Sample records for assembly heme synthesis

  1. Overexpression of the yeast frataxin homolog (Yfh1): contrasting effects on iron-sulfur cluster assembly, heme synthesis and resistance to oxidative stress

    DEFF Research Database (Denmark)

    Seguin, Alexandra; Bayot, Aurélien; Dancis, Andrew


    Friedreich's ataxia is generally associated with defects in [Fe-S] cluster assembly/stability and heme synthesis and strong susceptibility to oxidative stress. We used the yeast (Saccharomyces cerevisiae) model of Friedreich's ataxia to study the physiological consequences of modulating...

  2. Overexpression of the yeast frataxin homolog (Yfh1): contrasting effects on iron-sulfur cluster assembly, heme synthesis and resistance to oxidative stress. (United States)

    Seguin, Alexandra; Bayot, Aurélien; Dancis, Andrew; Rogowska-Wrzesinska, Adelina; Auchère, Françoise; Camadro, Jean-Michel; Bulteau, Anne-Laure; Lesuisse, Emmanuel


    Friedreich's ataxia is generally associated with defects in [Fe-S] cluster assembly/stability and heme synthesis and strong susceptibility to oxidative stress. We used the yeast (Saccharomyces cerevisiae) model of Friedreich's ataxia to study the physiological consequences of modulating the expression of the frataxin gene (YFH1). We show that the number of frataxin molecules per wild-type cell varies from less than 200 to 1500 according to the iron concentration in the medium. Cells overexpressing YFH1 on a plasmid (2muYFH1; about 3500 molecules Yfh1/cell) took up more iron than wild-type cells and displayed defective [Fe-S] cluster assembly/stability in vivo. By contrast, endogenous mitochondrial iron was more available to ferrochelatase in 2muYFH1 cells than in wild-type cells, resulting in higher levels of heme synthesis in vitro. Frataxin overproduction resulted in a shift from frataxin trimers to frataxin oligomers of higher molecular mass in the mitochondrial matrix. Much fewer carbonylated proteins were present in 2muYFH1 cells, and these cells were more resistant to oxidizing agents than wild-type cells, which probably resulted from the lower production of hydrogen peroxide by the mitochondria of 2muYFH1 cells compared to wild-type cells. To our knowledge, this work is the first description where major frataxin-related phenotypes ([Fe-S] cluster assembly and heme synthesis) can be split in vivo, suggesting that frataxin has independent roles in both processes, and that the optimal conditions for these independent roles are different.

  3. The heme a synthase Cox15 associates with cytochrome c oxidase assembly intermediates during Cox1 maturation. (United States)

    Bareth, Bettina; Dennerlein, Sven; Mick, David U; Nikolov, Miroslav; Urlaub, Henning; Rehling, Peter


    Cox1, the core subunit of the cytochrome c oxidase, receives two heme a cofactors during assembly of the 13-subunit enzyme complex. However, at which step of the assembly process and how heme is inserted into Cox1 have remained an enigma. Shy1, the yeast SURF1 homolog, has been implicated in heme transfer to Cox1, whereas the heme a synthase, Cox15, catalyzes the final step of heme a synthesis. Here we performed a comprehensive analysis of cytochrome c oxidase assembly intermediates containing Shy1. Our analyses suggest that Cox15 displays a role in cytochrome c oxidase assembly, which is independent of its functions as the heme a synthase. Cox15 forms protein complexes with Shy1 and also associates with Cox1-containing complexes independently of Shy1 function. These findings indicate that Shy1 does not serve as a mobile heme carrier between the heme a synthase and maturing Cox1 but rather cooperates with Cox15 for heme transfer and insertion in early assembly intermediates of cytochrome c oxidase.

  4. Molecular hijacking of siroheme for the synthesis of heme and d1 heme. (United States)

    Bali, Shilpa; Lawrence, Andrew D; Lobo, Susana A; Saraiva, Lígia M; Golding, Bernard T; Palmer, David J; Howard, Mark J; Ferguson, Stuart J; Warren, Martin J


    Modified tetrapyrroles such as chlorophyll, heme, siroheme, vitamin B(12), coenzyme F(430), and heme d(1) underpin a wide range of essential biological functions in all domains of life, and it is therefore surprising that the syntheses of many of these life pigments remain poorly understood. It is known that the construction of the central molecular framework of modified tetrapyrroles is mediated via a common, core pathway. Herein a further branch of the modified tetrapyrrole biosynthesis pathway is described in denitrifying and sulfate-reducing bacteria as well as the Archaea. This process entails the hijacking of siroheme, the prosthetic group of sulfite and nitrite reductase, and its processing into heme and d(1) heme. The initial step in these transformations involves the decarboxylation of siroheme to give didecarboxysiroheme. For d(1) heme synthesis this intermediate has to undergo the replacement of two propionate side chains with oxygen functionalities and the introduction of a double bond into a further peripheral side chain. For heme synthesis didecarboxysiroheme is converted into Fe-coproporphyrin by oxidative loss of two acetic acid side chains. Fe-coproporphyrin is then transformed into heme by the oxidative decarboxylation of two propionate side chains. The mechanisms of these reactions are discussed and the evolutionary significance of another role for siroheme is examined.

  5. Dietary iron controls circadian hepatic glucose metabolism through heme synthesis. (United States)

    Simcox, Judith A; Mitchell, Thomas Creighton; Gao, Yan; Just, Steven F; Cooksey, Robert; Cox, James; Ajioka, Richard; Jones, Deborah; Lee, Soh-Hyun; King, Daniel; Huang, Jingyu; McClain, Donald A


    The circadian rhythm of the liver maintains glucose homeostasis, and disruption of this rhythm is associated with type 2 diabetes. Feeding is one factor that sets the circadian clock in peripheral tissues, but relatively little is known about the role of specific dietary components in that regard. We assessed the effects of dietary iron on circadian gluconeogenesis. Dietary iron affects circadian glucose metabolism through heme-mediated regulation of the interaction of nuclear receptor subfamily 1 group d member 1 (Rev-Erbα) with its cosuppressor nuclear receptor corepressor 1 (NCOR). Loss of regulated heme synthesis was achieved by aminolevulinic acid (ALA) treatment of mice or cultured cells to bypass the rate-limiting enzyme in hepatic heme synthesis, ALA synthase 1 (ALAS1). ALA treatment abolishes differences in hepatic glucose production and in the expression of gluconeogenic enzymes seen with variation of dietary iron. The differences among diets are also lost with inhibition of heme synthesis with isonicotinylhydrazine. Dietary iron modulates levels of peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), a transcriptional activator of ALAS1, to affect hepatic heme. Treatment of mice with the antioxidant N-acetylcysteine diminishes PGC-1α variation observed among the iron diets, suggesting that iron is acting through reactive oxygen species signaling.

  6. Delayed globin synthesis leads to excess heme and the macrocytic anemia of Diamond Blackfan anemia and del(5q) myelodysplastic syndrome. (United States)

    Yang, Zhantao; Keel, Siobán B; Shimamura, Akiko; Liu, Li; Gerds, Aaron T; Li, Henry Y; Wood, Brent L; Scott, Bart L; Abkowitz, Janis L


    Diamond Blackfan anemia (DBA) and myelodysplastic syndrome (MDS) with isolated del(5q) are severe macrocytic anemias; although both are associated with impaired ribosome assembly, why the anemia occurs is not known. We cultured marrow cells from DBA (n = 3) and del(5q) MDS (n = 6) patients and determined how heme (a toxic chemical) and globin (a protein) are coordinated. We show that globin translation initiates slowly, whereas heme synthesis proceeds normally. This results in insufficient globin protein, excess heme and excess reactive oxygen species in early erythroid precursors, and CFU-E (colony-forming unit-erythroid)/proerythroblast cell death. The cells that can more rapidly and effectively export heme or can slow heme synthesis preferentially survive and appropriately mature. Consistent with these observations, treatment with 10 μM succinylacetone, a specific inhibitor of heme synthesis, improved the erythroid cell output of DBA and del(5q) MDS marrow cultures by 68 to 95% (P = 0.03 to 0.05), whereas the erythroid cell output of concurrent control marrow cultures decreased by 4 to 13%. Our studies demonstrate that erythropoiesis fails when heme exceeds globin. Our data further suggest that therapies that decrease heme synthesis (or facilitate heme export) could improve the red blood cell production of persons with DBA, del(5q) MDS, and perhaps other macrocytic anemias.

  7. Synthetic heme/copper assemblies: toward an understanding of cytochrome c oxidase interactions with dioxygen and nitrogen oxides. (United States)

    Hematian, Shabnam; Garcia-Bosch, Isaac; Karlin, Kenneth D


    Our long-time niche in synthetic biological inorganic chemistry has been to design ligands and generate coordination complexes of copper or iron ions or both, those reacting with dioxygen (O2) or nitrogen oxides (e.g., nitric oxide (NO(g)) and nitrite (NO2(-))) or both. As inspiration for this work, we turn to mitochondrial cytochrome c oxidase, which is responsible for dioxygen consumption and is also the predominant target for NO(g) and nitrite within mitochondria. In this Account, we highlight recent advances in studying synthetic heme/Cu complexes in two respects. First, there is the design, synthesis, and characterization of new O2 adducts whose further study will add insights into O2 reductive cleavage chemistry. Second, we describe how related heme/Cu constructs reduce nitrite ion to NO(g) or the reverse, oxidize NO(g) to nitrite. The reactions of nitrogen oxides occur as part of CcO's function, which is intimately tied to cellular O2 balance. We had first discovered that reduced heme/Cu compounds react with O2 giving μ-oxo heme-Fe(III)-O-Cu(II)(L) products; their properties are discussed. The O-atom is derived from dioxygen, and interrogations of these systems led to the construction and characterization of three distinctive classes of heme-peroxo complexes, two high-spin and one low-spin species. Recent investigations include a new approach to the synthesis of low-spin heme-peroxo-Cu complexes, employing a "naked" synthon, where the copper ligand denticity and geometric types can be varied. The result is a collection of such complexes; spectroscopic and structural features (by DFT calculations) are described. Some of these compounds are reactive toward reductants/protons effecting subsequent O-O cleavage. This points to how subtle improvements in ligand environment lead to a desired local structure and resulting optimized reactivity, as known to occur at enzyme active sites. The other sector of research is focused on heme/Cu assemblies mediating the redox

  8. Nitric oxide generation from heme/copper assembly mediated nitrite reductase activity. (United States)

    Hematian, Shabnam; Siegler, Maxime A; Karlin, Kenneth D


    Nitric oxide (NO) as a cellular signaling molecule and vasodilator regulates a range of physiological and pathological processes. Nitrite (NO2 (-)) is recycled in vivo to generate nitric oxide, particularly in physiologic hypoxia and ischemia. The cytochrome c oxidase binuclear heme a 3/CuB active site is one entity known to be responsible for conversion of cellular nitrite to nitric oxide. We recently reported that a partially reduced heme/copper assembly reduces nitrite ion, producing nitric oxide; the heme serves as the reductant and the cupric ion provides a Lewis acid interaction with nitrite, facilitating nitrite (N-O) bond cleavage (Hematian et al., J. Am. Chem. Soc. 134:18912-18915, 2012). To further investigate this nitrite reductase chemistry, copper(II)-nitrito complexes with tridentate and tetradentate ligands were used in this study, where either O,O'-bidentate or O-unidentate modes of nitrite binding to the cupric center are present. To study the role of the reducing ability of the ferrous heme center, two different tetraarylporphyrinate-iron(II) complexes, one with electron-donating para-methoxy peripheral substituents and the other with electron-withdrawing 2,6-difluorophenyl substituents, were used. The results show that differing modes of nitrite coordination to the copper(II) ion lead to differing kinetic behavior. Here, also, the ferrous heme is in all cases the source of the reducing equivalent required to convert nitrite to nitric oxide, but the reduction ability of the heme center does not play a key role in the observed overall reaction rate. On the basis of our observations, reaction mechanisms are proposed and discussed in terms of heme/copper heterobinuclear structures.

  9. Molecular hijacking of siroheme for the synthesis of heme and d1 heme


    Bali, Shilpa; Lawrence, Andrew D.; Lobo, Susana A; Saraiva, Lígia M.; Golding, Bernard T.; Palmer, David J.; Mark J. Howard; Ferguson, Stuart J.; Warren, Martin J.


    Modified tetrapyrroles such as chlorophyll, heme, siroheme, vitamin B12, coenzyme F430, and heme d1 underpin a wide range of essential biological functions in all domains of life, and it is therefore surprising that the syntheses of many of these life pigments remain poorly understood. It is known that the construction of the central molecular framework of modified tetrapyrroles is mediated via a common, core pathway. Herein a further branch of the modified tetrapyrrole biosynthesis pathway i...

  10. Gold nanoparticle assisted assembly of a heme protein for enhancement of long-range interfacial electron transfer

    DEFF Research Database (Denmark)

    Jensen, Palle Skovhus; Chi, Qijin; Grumsen, Flemming Bjerg


    of bioelectronics. A key challenge in molecular bioelectronics is to improve the efficiency of long-range charge transfer. The present work shows that this can be achieved by nanoparticle (NP) assisted assembly of cytochrome c (cyt c) on macroscopic single-crystalline electrode surfaces. We present the synthesis...... and characterization of water-soluble gold nanoparticles (AuNPs) with core diameter 3-4 nm and their application for the enhancement of long-range interfacial ET of a heme protein. Gold nanoparticles were electrostatically conjugated with cyt c to form nanoparticle-protein hybrid ET systems with well......-defined stoichiometry. The systems were investigated in homogeneous solution and at liquid/solid interface. Conjugation of cyt c results in a small but consistent broadening of the nanoparticle plasmon band. This phenomenon can be explained in terms of long-range electronic interactions between the gold nanoparticle...

  11. Identification and phylogenetic analysis of heme synthesis genes in trypanosomatids and their bacterial endosymbionts.

    Directory of Open Access Journals (Sweden)

    João M P Alves

    Full Text Available It has been known for decades that some insect-infecting trypanosomatids can survive in culture without heme supplementation while others cannot, and that this capability is associated with the presence of a betaproteobacterial endosymbiont in the flagellate's cytoplasm. However, the specific mechanisms involved in this process remained obscure. In this work, we sequence and phylogenetically analyze the heme pathway genes from the symbionts and from their hosts, as well as from a number of heme synthesis-deficient Kinetoplastida. Our results show that the enzymes responsible for synthesis of heme are encoded on the symbiont genomes and produced in close cooperation with the flagellate host. Our evidence suggests that this synergistic relationship is the end result of a history of extensive gene loss and multiple lateral gene transfer events in different branches of the phylogeny of the Trypanosomatidae.

  12. Comparative Genomics of the Genus Porphyromonas Identifies Adaptations for Heme Synthesis within the Prevalent Canine Oral Species Porphyromonas cangingivalis. (United States)

    O'Flynn, Ciaran; Deusch, Oliver; Darling, Aaron E; Eisen, Jonathan A; Wallis, Corrin; Davis, Ian J; Harris, Stephen J


    Porphyromonads play an important role in human periodontal disease and recently have been shown to be highly prevalent in canine mouths. Porphyromonas cangingivalis is the most prevalent canine oral bacterial species in both plaque from healthy gingiva and plaque from dogs with early periodontitis. The ability of P. cangingivalis to flourish in the different environmental conditions characterized by these two states suggests a degree of metabolic flexibility. To characterize the genes responsible for this, the genomes of 32 isolates (including 18 newly sequenced and assembled) from 18 Porphyromonad species from dogs, humans, and other mammals were compared. Phylogenetic trees inferred using core genes largely matched previous findings; however, comparative genomic analysis identified several genes and pathways relating to heme synthesis that were present in P. cangingivalis but not in other Porphyromonads. Porphyromonas cangingivalis has a complete protoporphyrin IX synthesis pathway potentially allowing it to synthesize its own heme unlike pathogenic Porphyromonads such as Porphyromonas gingivalis that acquire heme predominantly from blood. Other pathway differences such as the ability to synthesize siroheme and vitamin B12 point to enhanced metabolic flexibility for P. cangingivalis, which may underlie its prevalence in the canine oral cavity.

  13. Engineering the assembly of heme cofactors in man-made proteins. (United States)

    Solomon, Lee A; Kodali, Goutham; Moser, Christopher C; Dutton, P Leslie


    Timely ligation of one or more chemical cofactors at preselected locations in proteins is a critical preamble for catalysis in many natural enzymes, including the oxidoreductases and allied transport and signaling proteins. Likewise, ligation strategies must be directly addressed when designing oxidoreductase and molecular transport functions in man-made, first-principle protein constructs intended to operate in vitro or in vivo. As one of the most common catalytic cofactors in biology, we have chosen heme B, along with its chemical analogues, to determine the kinetics and barriers to cofactor incorporation and bishistidine ligation in a range of 4-α-helix proteins. We compare five elementary synthetic designs (maquettes) and the natural cytochrome b562 that differ in oligomeric forms, apo- and holo-tertiary structural stability; qualities that we show can either assist or hinder assembly. The cofactor itself also imposes an assembly barrier if amphiphilicity ranges toward too hydrophobic or hydrophilic. With progressive removal of identified barriers, we achieve maquette assembly rates as fast as native cytochrome b562, paving the way to in vivo assembly of man-made hemoprotein maquettes and integration of artificial proteins into enzymatic pathways.

  14. A Heme-responsive Regulator Controls Synthesis of Staphyloferrin B in Staphylococcus aureus. (United States)

    Laakso, Holly A; Marolda, Cristina L; Pinter, Tyler B; Stillman, Martin J; Heinrichs, David E


    Staphylococcus aureus possesses a multitude of mechanisms by which it can obtain iron during growth under iron starvation conditions. It expresses an effective heme acquisition system (the iron-regulated surface determinant system), it produces two carboxylate-type siderophores staphyloferrin A and staphyloferrin B (SB), and it expresses transporters for many other siderophores that it does not synthesize. The ferric uptake regulator protein regulates expression of genes encoding all of these systems. Mechanisms of fine-tuning expression of iron-regulated genes, beyond simple iron regulation via ferric uptake regulator, have not been uncovered in this organism. Here, we identify the ninth gene of the sbn operon, sbnI, as encoding a ParB/Spo0J-like protein that is required for expression of genes in the sbn operon from sbnD onward. Expression of sbnD-I is drastically decreased in an sbnI mutant, and the mutant does not synthesize detectable SB during early phases of growth. Thus, SB-mediated iron acquisition is impaired in an sbnI mutant strain. We show that the protein forms dimers and tetramers in solution and binds to DNA within the sbnC coding region. Moreover, we show that SbnI binds heme and that heme-bound SbnI does not bind DNA. Finally, we show that providing exogenous heme to S. aureus growing in an iron-free medium results in delayed synthesis of SB. This is the first study in S. aureus that identifies a DNA-binding regulatory protein that senses heme to control gene expression for siderophore synthesis.

  15. Heme oxygenase activity modulates vascular endothelial growth factor synthesis in vascular smooth muscle cells. (United States)

    Dulak, Jozef; Józkowicz, Alicja; Foresti, Roberta; Kasza, Aneta; Frick, Matthias; Huk, Ihor; Green, Colin J; Pachinger, Otmar; Weidinger, Franz; Motterlini, Roberto


    Hypoxia, cytokines, and nitric oxide (NO) stimulate the generation of vascular endothelial growth factor (VEGF) and induce heme oxygenase-1 (HO-1) expression in vascular tissue. HO-1 degrades heme to carbon monoxide (CO), iron, and biliverdin, the latter being reduced to bilirubin by biliverdin reductase. In the present study, we investigated the role of HO-1 in the modulation of VEGF synthesis in rat vascular smooth muscle cells (VSMC). In VSMC stimulated with cytokines, inhibition of NO production significantly, but not completely, reduced VEGF release. In contrast, inhibition of HO activity by tin protoporphyrin IX (SnPPIX) totally prevented cytokine-induced increase in VEGF, despite an augmented synthesis of intracellular NO. Stimulation of HO-1 activity by hemin enhanced VEGF production; this effect was abrogated by blockade of the HO pathway. Similarly, VEGF synthesis induced by hypoxia was down-regulated by SnPPIX, but not by inhibitors of NO synthase. To elucidate further a direct involvement of HO-1 in the observed effects, we generated transfected cells that overexpressed the HO-1 gene. Notably, these cells synthesized significantly more VEGF protein than cells transfected with a control gene. Among the products of HO-1, biliverdin and bilirubin showed no effect, whereas iron ions inhibited VEGF synthesis. Exposure of cells to 1% CO resulted in a marked accumulation of VEGF (20-fold increase) over the basal level. Our data indicate that HO-1 activity influences the generation of VEGF in VSMC in both normoxic and hypoxic conditions. As CO and iron, respectively the inducer and the inhibitor of VEGF synthesis, are concomitantly produced during the degradation of heme, these data indicate that HO by-products may differentially modulate VEGF production.

  16. Molecular basis of inherited microcytic anemia due to defects in iron acquisition or heme synthesis. (United States)

    Iolascon, Achille; De Falco, Luigia; Beaumont, Carole


    Microcytic anemia is the most commonly encountered anemia in general medical practice. Nutritional iron deficiency and beta thalassemia trait are the primary causes in pediatrics, whereas bleeding disorders and anemia of chronic disease are common in adulthood. Microcytic hypochromic anemia can result from a defect in globin genes, in heme synthesis, in iron availability or in iron acquisition by the erythroid precursors. These microcytic anemia can be sideroblastic or not, a trait which reflects the implications of different gene abnormalities. Iron is a trace element that may act as a redox component and therefore is integral to vital biological processes that require the transfer of electrons as in oxygen transport, oxidative phosphorylation, DNA biosynthesis and xenobiotic metabolism. However, it can also be pro-oxidant and to avoid its toxicity, iron metabolism is strictly controlled and failure of these control systems could induce iron overload or iron deficient anemia. During the past few years, several new discoveries mostly arising from human patients or mouse models have highlighted the implication of iron metabolism components in hereditary microcytic anemia, from intestinal absorption to its final inclusion into heme. In this paper we will review the new information available on the iron acquisition pathway by developing erythrocytes and its regulation, and we will consider only inherited microcytosis due to heme synthesis or to iron metabolism defects. This information could be useful in the diagnosis and classification of these microcytic anemias.

  17. Cloning and expression of zebrafish genes encoding the heme synthesis enzymes uroporphyrinogen III synthase (UROS) and protoporphyrinogen oxidase (PPO). (United States)

    Hanaoka, Ryuki; Dawid, Igor B; Kawahara, Atsuo


    Heme is synthesized from glycine and succinyl CoA by eight heme synthesis enzymes. Although genetic defects in any of these enzymes are known to cause severe human blood diseases, their developmental expression in mammals is unknown. In this paper, we report two zebrafish heme synthesis enzymes, uroporphyrinogen III synthase (UROS) and protoporphyrinogen oxidase (PPO) that are well conserved in comparison to their human counterparts. Both UROS and PPO formed pairs of bilateral stripes in the lateral plate mesoderm at the 15-somite stage. At 24 h post-fertilization (hpf), UROS and PPO were predominantly expressed in the intermediate cell mass (ICM) that is the major site of primitive hematopoiesis. The expression of UROS and PPO was drastically suppressed in the bloodless mutants cloche and vlad tepes/gata 1 from 15-somite to 24hpf stages, indicating that both cloche and vlad tepes/gata 1 are required for the induction and maintenance of UROS and PPO expression in the ICM.

  18. [The process of heme synthesis in bone marrow mesenchymal stem cells cultured under fibroblast growth factor bFGF and hypoxic conditions]. (United States)

    Poleshko, A G; Lobanok, E S; Mezhevikina, L M; Fesenko, E E; Volotkovskiĭ, I D


    It was demonstrated that fibroblast growth factor bFGF influences the process of heme synthesis, the proliferation activity and viability of bone marrow mesenchymal stem cells in culture under hypoxic conditions. The addition of fibroblast growth factor bFGF (7 ng/ml) to the medium under above conditions led to the accumulation of aminolevulinic acid--an early porphyrin and heme precursor, an increase in CD 71 expression--a transferrin receptor, and also a decrease in porphyrin pigments and heme contents--a late precursor and end products of heme synthesis, respectively. It was found that cultivation of the cells under hypoxic conditions and bFGF is an optimum to maintain high viability and proliferation capacity of the mesenchymal stem cells.

  19. Measurement of heme concentration. (United States)

    Sinclair, P R; Gorman, N; Jacobs, J M


    Heme (iron protoporphyrin IX) is a prosthetic group for a number of hemoproteins in different tissues (e.g., hemoglobin, myoglobin, cytochrome P-450s, mitochondrial cytochromes, catalases, and peroxidases). Mutations in the biosynthetic pathway can affect the synthesis and/or degradation of heme. Several assays are provided in this unit for quantifying heme: a spectrophotometric assay based on the characteristic absorption spectrum of oxidized and reduced form of the hemochrome formed by replacing the nitrogen ligands with pyridine; a fluorescence assay based on removal of the iron by a heated, strong oxalic acid solution to produce fluorescent protoporphyrin; a reversed-phase HPLC assay to measure heme and intermediates in the synthetic pathway; and a radiometric assay to measure newly synthesized heme in tissue culture cells.

  20. Diagnosis and treatment of sideroblastic anemias: from defective heme synthesis to abnormal RNA splicing. (United States)

    Cazzola, Mario; Malcovati, Luca


    The sideroblastic anemias are a heterogeneous group of inherited and acquired disorders characterized by the presence of ring sideroblasts in the bone marrow. X-linked sideroblastic anemia (XLSA) is caused by germline mutations in ALAS2. Hemizygous males have a hypochromic microcytic anemia, which is generally mild to moderate and is caused by defective heme synthesis and ineffective erythropoiesis. XLSA is a typical iron-loading anemia; although most patients are responsive to pyridoxine, treatment of iron overload is also important in the management of these patients. Autosomal recessive sideroblastic anemia attributable to mutations in SLC25A38, a member of the mitochondrial carrier family, is a severe disease: patients present in infancy with microcytic anemia, which soon becomes transfusion dependent. Conservative therapy includes regular red cell transfusion and iron chelation, whereas allogenic stem cell transplantation represents the only curative treatment. Refractory anemia with ring sideroblasts (RARS) is a myelodysplastic syndrome characterized mainly by anemia attributable to ineffective erythropoiesis. The clinical course of RARS is generally indolent, but there is a tendency to worsening of anemia over time, so that most patients become transfusion dependent in the long run. More than 90% of these patients carry somatic mutations in SF3B1, a gene encoding a core component of the RNA splicing machinery. These mutations cause misrecognition of 3' splice sites in downstream genes, resulting in truncated gene products and/or decreased expression attributable to nonsense-mediated RNA decay; this explains the multifactorial pathogenesis of RARS. Variants of RARS include refractory cytopenia with multilineage dysplasia and ring sideroblasts, and RARS associated with marked thrombocytosis; these variants involve additional genetic lesions. Inhibitors of molecules of the transforming growth factor-β superfamily have been shown recently to target ineffective

  1. Synthesis, assembly and device of 1-dimentional nanostructures

    Institute of Scientific and Technical Information of China (English)


    Synthesis and assembly of 1-dimentional (1-D) nanostructures and measurement of their electrical and optical properties are very important in fabrication of nanodevices. Recent developments in this field are summarized in this review. The assembling methods can be divided into two classes: assembly using macroscopic field forces and microfluidic-assisted-template-integration. The former can assemble nanowires by controlling direction and intensity of electric or magnetic field, while the latter represents a general assembly strategy for any kind of 1-D nanostructures. The assembly of 1-D nanostructures will make it possible to fabricate nanosensors, nanolasers and nanoscale logic gate circuits for computation.

  2. Measurement of heme efflux and heme content in isolated developing chloroplasts. [Cucumis sativus, cv. Sumter

    Energy Technology Data Exchange (ETDEWEB)

    Thomas, J.; Weinstein, J.D. (Clemson Univ., SC (USA))


    Hemes destined for cytosolic hemoproteins must originate in one of the cellular compartments which have the capacity for heme synthesis, namely the chloroplast or the mitochondria. Since developing chloroplasts from greening cucumber (Cucumis sativus, cv. Sumter) cotyledons are known to contain complete heme and chlorophyll biosynthetic pathways, they were tested for their capacity export hemes. Picomole quantities of heme were measured by reconstitution of the heme with apo-peroxidase and subsequent determination of peroxidase activity. The assay method was sensitive (as little as 0.7 picomole of heme could be detected in a volume of 100 microliters) and was linear with heme concentration. When intact plastids were incubated with apo-peroxidase, a steady-state rate of efflux between 0.12 and 0.45 picomole heme/minute/milligram plastid protein was measured. The efflux rate was not due to plastid breakage and could be enhanced by incubating with the heme precursor, {delta}-aminolevulinic acid. Cold acetone extraction removed 47 {plus minus} 17 picomoles heme/milligram plastid protein from the total b-type heme pool in the chloroplasts (166 {plus minus} 9 picomoles heme/milligram protein, by acid-acetone extraction). The reconstitution technique provided a similar estimate of readily exchangeable heme in the plastid, 37 {plus minus} 8 picomoles heme/milligram protein (or 6 micromolar in the plastids). These values may be indicative of a free heme pool which exists in the chloroplast.

  3. Functional Characterization of the Canine Heme-Regulated eIF2α Kinase: Regulation of Protein Synthesis

    Directory of Open Access Journals (Sweden)

    Kimon C. Kanelakis


    Full Text Available The heme-regulated inhibitor (HRI negatively regulates protein synthesis by phosphorylating eukaryotic initiation factor-2α (eIF2α thereby inhibiting protein translation. The importance of HRI in regulating hemoglobin synthesis in erythroid cells makes it an attractive molecular target in need of further characterization. In this work, we have cloned and expressed the canine form of the HRI kinase. The canine nucleotide sequence has 86%, 82%, and 81% identity to the human, mouse, and rat HRI, respectively. It was noted that an isoleucine residue in the ATP binding site of human, rat, and mouse HRI is replaced by a valine in the canine kinase. The expression of canine HRI protein by in vitro translation using wheat germ lysate or in Sf9 cells using a baculovirus expression system was increased by the addition of hemin. Following purification, the canine protein was found to be 72 kD and showed kinase activity determined by its ability to phosphorylate a synthetic peptide substrate. Quercetin, a kinase inhibitor known to inhibit mouse and human HRI, inhibits canine HRI in a concentration-dependent manner. Additionally, quercetin is able to increase de novo protein synthesis in canine reticulocytes. We conclude that the canine is a suitable model species for studying the role of HRI in erythropoiesis.

  4. Arsenite alters heme synthesis in long-term cultures of adult rat hepatocytes. (United States)

    Aguilar-González, M G; Hernández, A; López, M L; Mendoza-Figueroa, T; Albores, A


    Arsenite (As[III]) effects on the intermediate steps of heme biosynthesis were studied in adult rat hepatocytes seeded on a feeder layer of 3T3 cells (3T3-hepatocytes) and maintained for 2 weeks with culture medium non-supplemented or supplemented with 150 microM 5-aminolevulinic acid (ALA). The activities of the intracellular enzymes porphobilinogen deaminase (PBG-D), uroporphyrinogen III synthase (UROIII-S), and uroporphyrinogen III decarboxylase (URO-D), and the intermediary uroporphyrins (URO), coproporphyrins (COPRO) and protoporphyrin IX (PROTO) were determined in these cultures. The 3T3-hepatocytes maintained the activities of PBG-D, UROIII-S and URO-D during 2 weeks and ALA addition to the culture medium increased PBG-D (2-3-fold) and UROIII-S (50%) activities and porphyrin production, which accumulated as PROTO. Exposure to 3.9 microM As(III) inhibited UROIII-S activity (down to 34%), and PBG-D and URO-D activities to a lower extent; these effects were magnified by ALA supplementation. As(III) also produced an intracellular accumulation and a decreased excretion of PROTO, and a 31% reduction of the COPRO/URO ratio in the culture medium. Additionally, As(III) caused cytoplasmic vacuolization and lipid accumulation. Our results show that As(III) exposure selectively inhibits several intermediary enzymes of heme metabolism and affects the intra- and extracellular content of porphyrins and their ratio in the culture medium. They also confirm that 3T3-hepatocytes are a suitable in vitro model to study hepatic heme metabolism and its alterations by hepatotoxic chemicals.

  5. Impact of heme oxygenase-1 on cholesterol synthesis, cholesterol efflux and oxysterol formation in cultured astroglia. (United States)

    Hascalovici, Jacob R; Song, Wei; Vaya, Jacob; Khatib, Soliman; Fuhrman, Bianca; Aviram, Michael; Schipper, Hyman M


    Up-regulation of heme oxygenase-1 (HO-1) and altered cholesterol (CH) metabolism are characteristic of Alzheimer-diseased neural tissues. The liver X receptor (LXR) is a molecular sensor of CH homeostasis. In the current study, we determined the effects of HO-1 over-expression and its byproducts iron (Fe(2+)), carbon monoxide (CO) and bilirubin on CH biosynthesis, CH efflux and oxysterol formation in cultured astroglia. HO-1/LXR interactions were also investigated in the context of CH efflux. hHO-1 over-expression for 3 days ( approximately 2-3-fold increase) resulted in a 30% increase in CH biosynthesis and a two-fold rise in CH efflux. Both effects were abrogated by the competitive HO inhibitor, tin mesoporphyrin. CO, released from administered CORM-3, significantly enhanced CH biosynthesis; a combination of CO and iron stimulated CH efflux. Free iron increased oxysterol formation three-fold. Co-treatment with LXR antagonists implicated LXR activation in the modulation of CH homeostasis by heme degradation products. In Alzheimer's disease and other neuropathological states, glial HO-1 induction may transduce ambient noxious stimuli (e.g. beta-amyloid) into altered patterns of glial CH homeostasis. As the latter may impact synaptic plasticity and neuronal repair, modulation of glial HO-1 expression (by pharmacological or other means) may confer neuroprotection in patients with degenerative brain disorders.

  6. Effect of prostaglandin-J(2) on VEGF synthesis depends on the induction of heme oxygenase-1. (United States)

    Jozkowicz, Alicja; Huk, Ihor; Nigisch, Anneliese; Weigel, Günter; Weidinger, Franz; Dulak, Jozef


    Heme oxygenase-1 (HO-1) is an inducible enzyme that degrades heme to carbon monoxide, iron ions, and biliverdin. Its expression can be induced by 15-deoxy-Delta(12,14)prostaglandin-J(2) (15d-PGJ(2)), a natural ligand of peroxisome proliferator-activated receptor-gamma transcription factor. In macrophages and vascular smooth muscle cells, 15d-PGJ(2) up-regulates the expression of vascular endothelial growth factor (VEGF), a fundamental regulator of angiogenesis. Here we investigated the involvement of HO-1 in the 15d-PGJ(2)-mediated regulation of VEGF production by human microvascular endothelial cells (HMEC-1). Resting HMEC-1 released approximately 20 pg/ml VEGF protein after 24 h of incubation. Treatment of cells with 15d-PGJ(2) (1-10 microM) significantly and dose-dependently increased the VEGF promoter activity, mRNA expression, and protein secretion. In the same cells, 15d-PGJ(2) potently induced the expression of HO-1 protein that correlated with HO-1 promoter activity. Activation of HO-1 with hemin or ectopic overexpression of HO-1 in HMEC-1 perfectly mimicked the effect of 15d-PGJ(2) and led to increased VEGF production. Importantly, the inhibition of the HO-1 pathway by tin protoporphyrin-IX significantly reduced the stimulatory effect of 15d-PGJ(2) on VEGF synthesis. Thus, we postulate that the up-regulation of VEGF expression in response to 15d-PGJ(2 )in HMEC-1 is mediated by the activation of HO-1.

  7. A critical role for the co-repressor N-CoR in erythroid differentiation and heme synthesis

    Institute of Scientific and Technical Information of China (English)

    Dianzheng Zhang; Ellen Cho; Jiemin Wong


    Co-repressor N-CoR (nuclear receptor co-repressor) has important roles in different biological processes, including proliferation, differentiation and development. Mutant mice lacking N-CoR are embryonically lethal and appear to die from anemia owing to defects in definitive erythropoiesis. However, the underlying molecular mechanisms of N-CoR-mediated erythroid differentiation are largely unknown. Using the human erythroleukemic K562 cell line, which can be chemically induced to differentiate into either erythroid or megakaryocytic lineages depending on the inducers used, we have investigated the role of N-CoR in erythroid differentiation. We show that knockdown of N-CoR either transiently (siRNA) or permanently (shRNA) impairs the cytosine arabinoside (Ara-C)- but not hemin-induced erythroid differentiation of K562 cells. RT-PCR analysis reveals that N-CoR is required for induction by Ara-C of 5-aminolevulinate synthase (ALA-S2), a key enzyme involved in heme biosynthesis. Furthermore, the amount of N-CoR proteins increases significantly during Ara-C-induced K562 differentiation, apparently through a post-transcriptional mechanism. Consistent with the data from N-CoR-null mice, N-CoR is not required for the differentiation of K562 cells into megakaryocytic lineages, induced by phorbol 12-myristate 13-acetate. Thus, our in vitro study confirms a role for N-CoR in erythroid differentiation and reveals for the first time that N-CoR is required for the induction of a key enzyme involved in heme synthesis.

  8. Synthesis and Evaluation of Amyloid β Derived and Amyloid β Independent Enhancers of the Peroxidase-like Activity of Heme. (United States)

    Wißbrock, Amelie; Kühl, Toni; Silbermann, Katja; Becker, Albert J; Ohlenschläger, Oliver; Imhof, Diana


    Labile heme has been suggested to have an impact in several severe diseases. In the context of Alzheimer's disease (AD), however, decreased levels of free heme have been reported. Therefore, we were looking for an assay system that can be used for heme concentration determination. From a biochemical point of view the peroxidase activity of the Aβ-heme complex seemed quite attractive to pursue this goal. As a consequence, a peptide that is able to increase the readout even in the case of a low heme concentration is favorable. The examination of Aβ- and non-Aβ-derived peptides in complex with heme revealed that the peroxidase-like activity significantly depends on the peptide sequence and length. A 23mer His-based peptide derived from human fatty acyl-CoA reductase 1 in complex with heme exhibited a significantly higher peroxidase activity than Aβ(40)-heme. Structural modeling of both complexes demonstrated that heme binding via a histidine can be supported by hydrogen bond interactions of a basic residue near the propionate carboxyl function of protoporphyrin IX. Furthermore, the interplay of Aβ-heme and the lipoprotein LDL as a potential physiological effector of Aβ was examined.

  9. A functional mimic of natural peroxidases : synthesis and catalytic activity of a non-heme iron/peptide hydroperoxide complex

    NARCIS (Netherlands)

    Choma, Christin T.; Schudde, Ebe P.; Kellogg, Richard M.; Robillard, George T.; Feringa, Ben L.


    Site-selective attachment of unprotected peptides to a non-heme iron complex is achieved by displacing two halides on the catalyst by peptide caesium thiolates. This coupling approach should be compatible with any peptide sequence provided there is only a single reduced cysteine. The oxidation activ

  10. Synthesis and assembly of self-complementary calix[4]arenes. (United States)

    Shimizu, K D; Rebek, J


    A calix[4]arene was designed to reversibly dimerize and form an egg-shaped enclosure. Adhesive interactions in the assembly were provided by four self-associating ureas, which form a cyclic array containing 16 hydrogen bonds. The synthesis was completed in four steps from the previously described O,O',O",O"'-tetrabenzylcalix[4]arene. Evidence for dimerization of the calixarene tetraurea was provided by H NMR, mass spectrometry, and the observation of encapsulated molecules. The resulting cavity was of sufficient size to capture guests such as ethyl benzene and p-xylene. Images Fig. 1 Fig. 2 Fig. 6 PMID:8618910

  11. The Structure of the Complex between Yeast Frataxin and Ferrochelatase: CHARACTERIZATION AND PRE-STEADY STATE REACTION OF FERROUS IRON DELIVERY AND HEME SYNTHESIS. (United States)

    Söderberg, Christopher; Gillam, Mallory E; Ahlgren, Eva-Christina; Hunter, Gregory A; Gakh, Oleksandr; Isaya, Grazia; Ferreira, Gloria C; Al-Karadaghi, Salam


    Frataxin is a mitochondrial iron-binding protein involved in iron storage, detoxification, and delivery for iron sulfur-cluster assembly and heme biosynthesis. The ability of frataxin from different organisms to populate multiple oligomeric states in the presence of metal ions, e.g. Fe(2+) and Co(2+), led to the suggestion that different oligomers contribute to the functions of frataxin. Here we report on the complex between yeast frataxin and ferrochelatase, the terminal enzyme of heme biosynthesis. Protein-protein docking and cross-linking in combination with mass spectroscopic analysis and single-particle reconstruction from negatively stained electron microscopic images were used to verify the Yfh1-ferrochelatase interactions. The model of the complex indicates that at the 2:1 Fe(2+)-to-protein ratio, when Yfh1 populates a trimeric state, there are two interaction interfaces between frataxin and the ferrochelatase dimer. Each interaction site involves one ferrochelatase monomer and one frataxin trimer, with conserved polar and charged amino acids of the two proteins positioned at hydrogen-bonding distances from each other. One of the subunits of the Yfh1 trimer interacts extensively with one subunit of the ferrochelatase dimer, contributing to the stability of the complex, whereas another trimer subunit is positioned for Fe(2+) delivery. Single-turnover stopped-flow kinetics experiments demonstrate that increased rates of heme production result from monomers, dimers, and trimers, indicating that these forms are most efficient in delivering Fe(2+) to ferrochelatase and sustaining porphyrin metalation. Furthermore, they support the proposal that frataxin-mediated delivery of this potentially toxic substrate overcomes formation of reactive oxygen species.

  12. Synthesis and self-assembly of complex hollow materials

    KAUST Repository

    Zeng, Hua Chun


    Hollow materials with interiors or voids and pores are a class of lightweight nanostructured matters that promise many future technological applications, and they have received significant research attention in recent years. On the basis of well-known physicochemical phenomena and principles, for example, several solution-based protocols have been developed for the general preparation of these complex materials under mild reaction conditions. This article is thus a short introductory review on the synthetic aspects of this field of development. The synthetic methodologies can be broadly divided into three major categories: (i) template-assisted synthesis, (ii) self-assembly with primary building blocks, and (iii) induced matter relocations. In most cases, both synthesis and self-assembly are involved in the above processes. Further combinations of these methodologies appear to be very important, as they will allow one to prepare functional materials at a higher level of complexity and precision. The synthetic strategies are introduced through some simple case studies with schematic illustrations. Salient features of the methods developed have been summarized, and some urgent issues of this field have also been indicated. © 2011 The Royal Society of Chemistry.

  13. Heme oxygenase: evolution, structure, and mechanism. (United States)

    Wilks, Angela


    Heme oxygenase has evolved to carry out the oxidative cleavage of heme, a reaction essential in physiological processes as diverse as iron reutilization and cellular signaling in mammals, synthesis of essential light-harvesting pigments in cyanobacteria and higher plants, and the acquisition of iron by bacterial pathogens. In all of these processes, heme oxygenase has evolved a similar structural and mechanistic scaffold to function within seemingly diverse physiological pathways. The heme oxygenase reaction is catalytically distinct from that of other hemoproteins such as the cytochromes P450, peroxidases, and catalases, but shares a hemoprotein scaffold that has evolved to generate a distinct activated oxygen species. In the following review we discuss the evolution of the structural and functional properties of heme oxygenase in light of the recent crystal structures of the mammalian and bacterial enzymes.

  14. Synthesis and Self-Assembly of Triangulenium Salts

    DEFF Research Database (Denmark)

    Shi, Dong

    This thesis describes the design and synthesis of asymmetrically substituted amphiphilic tis(dialkylamino)trioxiatriangulenium (ATOTA+) salts with different counter ions. Attention was focused on exploring the assembling properties of the ATOTA+ salts in aqueous media. A direct vortexing....... Addition of soft counter ion into the nanosheets solution could induce gluing of the nanosheets. The solid thin film formed from the formed nanosheets after water evaporation showed crystalline patterning order as revealed by x-ray diffraction (XRD) measurements. Chpater 5 reports the counter ion effect...... in our group. The motivation of this project is to find potential application of the fluorescence sensor in biolabeling and bioimaging techniques. Chapter 8 is a brief summary of the thesis....

  15. Dendrimers in Layer-by-Layer Assemblies: Synthesis and Applications

    Directory of Open Access Journals (Sweden)

    Katsuhiko Sato


    Full Text Available We review the synthesis of dendrimer-containing layer-by-layer (LbL assemblies and their applications, including biosensing, controlled drug release, and bio-imaging. Dendrimers can be built into LbL films and microcapsules by alternating deposition of dendrimers and counter polymers on the surface of flat substrates and colloidal microparticles through electrostatic bonding, hydrogen bonding, covalent bonding, and biological affinity. Dendrimer-containing LbL assemblies have been used to construct biosensors, in which electron transfer mediators and metal nanoparticles are often coupled with dendrimers. Enzymes have been successfully immobilized on the surface of electrochemical and optical transducers by forming enzyme/dendrimer LbL multilayers. In this way, high-performance enzyme sensors are fabricated. In addition, dendrimer LbL films and microcapsules are useful for constructing drug delivery systems because dendrimers bind drugs to form inclusion complexes or the dendrimer surface is covalently modified with drugs. Magnetic resonance imaging of cancer cells by iron oxide nanoparticles coated with dendrimer LbL film is also discussed.

  16. Synthesis and metrology of conducting carbon nanotube assemblies (United States)

    Longson, Timothy Jay

    Since its discovery, the carbon nanotube (CNT) has been proposed as one of the ultimate materials for its electrical, thermal and mechanical properties due to its incredibly strong sp2 bonds, low defect density, and large aspect ratio. Many experimental results on individual CNTs have confirmed these outstanding theoretically predicted properties. However, scaling these properties to the macroscopic regime has proved to be challenging. This work focused on the synthesis and measurement of highly conducting, macroscopic, CNT assemblies. Scaling up the synthesis of vertically aligned multiwalled CNT (MWNT) forests was investigated through the development of a large, 100mm, wafer scale, cold wall chemical vapor deposition chamber. In addition to the synthesis, two distinct CNT assemblies have been investigated. A linear morphology where CNTs are strung in series for electrical transport (CNT wires) and a massively parallel 2D array of vertically aligned CNTs for Thermal Interface Material (TIM) applications. Poymer-CNT wire composites have been fabricated by developing a coaxial CNT core-polymer shell electrospinning technique. The core-shell interactions in this system have been studied by way of Hansen's solubility parameters. The most well defined CNT core was achieved using a core solvent that is semi-immiscible with the shell solution, yet still a solvent of the shell polymer. Electrical characterization of the resulting CNT core has shown a two orders of magnitude increase in conductivity over traditional, homogeneously mixed, electrospun CNT wires. A number of vertically aligned MWNT assemblies were studied for their thermal interface properties. Double-sided Silicon substrate (MWNT-Si-MWNT) TIM assemblies were characterized using a DC, 1D reference bar, thermal measurement technique. While attempts to control MWNT density via a micelle template technique produced only 'spaghetti like' CNTs, sputter deposited catalyst provided stark variations in array density

  17. Non-heme induction of heme oxygenase-1 does not alter cellular iron metabolism. (United States)

    Sheftel, Alex D; Kim, Sangwon F; Ponka, Prem


    The catabolism of heme is carried out by members of the heme oxygenase (HO) family. The products of heme catabolism by HO-1 are ferrous iron, biliverdin (subsequently converted to bilirubin), and carbon monoxide. In addition to its function in the recycling of hemoglobin iron, this microsomal enzyme has been shown to protect cells in various stress models. Implicit in the reports of HO-1 cytoprotection to date are its effects on the cellular handling of heme/iron. However, the limited amount of uncommitted heme in non-erythroid cells brings to question the source of substrate for this enzyme in non-hemolytic circumstances. In the present study, HO-1 was induced by either sodium arsenite (reactive oxygen species producer) or hemin or overexpressed in the murine macrophage-like cell line, RAW 264.7. Both of the inducers elicited an increase in active HO-1; however, only hemin exposure caused an increase in the synthesis rate of the iron storage protein, ferritin. This effect of hemin was the direct result of the liberation of iron from heme by HO. Cells stably overexpressing HO-1, although protected from oxidative stress, did not display elevated basal ferritin synthesis. However, these cells did exhibit an increase in ferritin synthesis, compared with untransfected controls, in response to hemin treatment, suggesting that heme levels, and not HO-1, limit cellular heme catabolism. Our results suggest that the protection of cells from oxidative insult afforded by HO-1 is not due to the catabolism of significant amounts of cellular heme as thought previously.

  18. Free heme and sickle hemoglobin polymerization (United States)

    Uzunova, Veselina V.

    This work investigates further the mechanism of one of the most interesting of the protein self-assembly systems---the polymerization of sickle hemoglobin and the role of free heme in it. Polymerization of sickle hemoglobin is the primary event in the pathology of a chronic hemolytic condition called sickle cell anemia with complex pathogenesis, unexplained variability and symptomatic treatment. Auto-oxidation develops in hemoglobin solutions exposed to room temperature and causes release of ferriheme. The composition of such solutions is investigated by mass spectrometry. Heme dimers whose amount corresponds to the initial amounts of heme released from the protein are followed. Differences in the dimer peak height are established for hemoglobin variants A, S and C and depending on the exposure duration. The effects of free heme on polymerization kinetics are studied. Growth rates and two characteristic parameters of nucleation are measured for stored Hb S. After dialysis of polymerizing solutions, no spherulites are detected at moderately high supersaturation and prolonged exposure times. The addition of 0.16-0.26 mM amounts of heme to dialyzed solutions leads to restoration of polymerization. The measured kinetic parameters have higher values compared to the ones before dialysis. The amount of heme in non-dialyzed aged solution is characterized using spectrophotometry. Three methods are used: difference in absorbance of dialyzed and non-dialyzed solutions, characteristic absorbance of heme-albumin complex and absorbance of non-dialyzed solutions with added potassium cyanide. The various approaches suggest the presence of 0.12 to 0.18 mM of free ferriheme in such solutions. Open questions are whether the same amounts of free heme are present in vivo and whether the same mechanism operates intracellulary. If the answer to those questions is positive, then removal of free heme from erythrocytes can influence their readiness to sickle.

  19. Simultaneous synthesis/assembly of anisotropic cake-shaped porphyrin particles toward colloidal microcrystals. (United States)

    Wang, Ting; Kuang, Minxuan; Jin, Feng; Cai, Jinhua; Shi, Lei; Zheng, Yongmei; Wang, Jingxia; Jiang, Lei


    The one-step synthesis/assembly of a cake-shaped porphyrin colloidal microcrystal with tailored height-diameter was demonstrated based on interfacial assembly and the water-droplet template. The as-fabricated anisotropic colloidal crystals showed special optic properties and enhanced optic-limiting behavior.

  20. Porphyrin and heme metabolism and the porphyrias. (United States)

    Bonkovsky, Herbert L; Guo, Jun-Tao; Hou, Weihong; Li, Ting; Narang, Tarun; Thapar, Manish


    Porphyrins and metalloporphyrins are the key pigments of life on earth as we know it, because they include chlorophyll (a magnesium-containing metalloporphyrin) and heme (iron protoporphyrin). In eukaryotes, porphyrins and heme are synthesized by a multistep pathway that involves eight enzymes. The first and rate-controlling step is the formation of delta-aminolevulinic acid (ALA) from glycine plus succinyl CoA, catalyzed by ALA synthase. Intermediate steps occur in the cytoplasm, with formation of the monopyrrole porphobilinogen and the tetrapyrroles hydroxymethylbilane and a series of porphyrinogens, which are serially decarboxylated. Heme is utilized chiefly for the formation of hemoglobin in erythrocytes, myoglobin in muscle cells, cytochromes P-450 and mitochondrial cytochromes, and other hemoproteins in hepatocytes. The rate-controlling step of heme breakdown is catalyzed by heme oxygenase (HMOX), of which there are two isoforms, called HMOX1 and HMOX2. HMOX breaks down heme to form biliverdin, carbon monoxide, and iron. The porphyrias are a group of disorders, mainly inherited, in which there are defects in normal porphyrin and heme synthesis. The cardinal clinical features are cutaneous (due to the skin-damaging effects of excess deposited porphyrins) or neurovisceral attacks of pain, sometimes with weakness, delirium, seizures, and the like (probably due mainly to neurotoxic effects of ALA). The treatment of choice for the acute hepatic porphyrias is intravenous heme therapy, which repletes a critical regulatory heme pool in hepatocytes and leads to downregulation of hepatic ALA synthase, which is a biochemical hallmark of all forms of acute porphyria in relapse.

  1. Heme and non-heme iron transporters in non-polarized and polarized cells

    Directory of Open Access Journals (Sweden)

    Yasui Yumiko


    Full Text Available Abstract Background Heme and non-heme iron from diet, and recycled iron from hemoglobin are important products of the synthesis of iron-containing molecules. In excess, iron is potentially toxic because it can produce reactive oxygen species through the Fenton reaction. Humans can absorb, transport, store, and recycle iron without an excretory system to remove excess iron. Two candidate heme transporters and two iron transporters have been reported thus far. Heme incorporated into cells is degraded by heme oxygenases (HOs, and the iron product is reutilized by the body. To specify the processes of heme uptake and degradation, and the reutilization of iron, we determined the subcellular localizations of these transporters and HOs. Results In this study, we analyzed the subcellular localizations of 2 isoenzymes of HOs, 4 isoforms of divalent metal transporter 1 (DMT1, and 2 candidate heme transporters--heme carrier protein 1 (HCP1 and heme responsive gene-1 (HRG-1--in non-polarized and polarized cells. In non-polarized cells, HCP1, HRG-1, and DMT1A-I are located in the plasma membrane. In polarized cells, they show distinct localizations: HCP1 and DMT1A-I are located in the apical membrane, whereas HRG-1 is located in the basolateral membrane and lysosome. 16Leu at DMT1A-I N-terminal cytosolic domain was found to be crucial for plasma membrane localization. HOs are located in smooth endoplasmic reticulum and colocalize with NADPH-cytochrome P450 reductase. Conclusions HCP1 and DMT1A-I are localized to the apical membrane, and HRG-1 to the basolateral membrane and lysosome. These findings suggest that HCP1 and DMT1A-I have functions in the uptake of dietary heme and non-heme iron. HRG-1 can transport endocytosed heme from the lysosome into the cytosol. These localization studies support a model in which cytosolic heme can be degraded by HOs, and the resulting iron is exported into tissue fluids via the iron transporter ferroportin 1, which is

  2. Synthesis of nanocrystals and nanocrystal self-assembly (United States)

    Chen, Zhuoying

    Chapter 1. A general introduction is presented on nanomaterials and nanoscience. Nanoparticles are discussed with respect to their structure and properties. Ferroelectric materials and nanoparticles in particular are highlighted, especially in the case of the barium titanate, and their potential applications are discussed. Different nanocrystal synthetic techniques are discussed. Nanoparticle superlattices, the novel "meta-materials" built from self-assembly at the nanoscale, are introduced. The formation of nanoparticle superlattices and the importance and interest of synthesizing these nanostructures is discussed. Chapter 2. Advanced applications for high k dielectric and ferroelectric materials in the electronics industry continues to demand an understanding of the underlying physics in decreasing dimensions into the nanoscale. The first part of this chapter presents the synthesis, processing, and electrical characterization of nanostructured thin films (thickness ˜100 nm) of barium titanate BaTiO3 built from uniform nanoparticles (properties. We observe the BaTiO3 nanocrystals crystallize with evidence of tetragonality. Electric field dependent polarization measurements show spontaneous polarization and hysteresis, indicating ferroelectric behavior for the BaTiO 3 nanocrystalline films with grain sizes in the range of 10--30 nm. Dielectric measurements of the films show dielectic constants in the range of 85--90 over the 1 kHz--100 kHz, with low loss. We present nanocrystals as initial building blocks for the preparation of thin films which exhibit uniform nanostructured morphologies and grain sizes. In the second part of this chapter, a nonhydrolytic alcoholysis route to study the preparation of well-crystallized size-tunable BaTiO3 nanocrystals is presented. Different surfactants of amines, carboxylic acids, and alcohols were used to study the effect of size and morphological control over the nanocrystals. Techniques including X-ray diffraction, transmission

  3. Molecular self-assembly and nanochemistry: A chemical strategy for the synthesis of nanostructures (United States)

    Whitesides, George M.; Mathias, John P.; Seto, Christopher T.


    Molecular self assembly is the spontaneous association of molecules under equilibrium conditions into stable, structurally well-defined aggregates joined by non-covalent bonds. Molecular self-assembly is ubiquitous in biological systems, and underlies the formation of a wide variety of complex biological structures. Understanding self-assembly and the associated non-covalent interactions that connect complementary interacting molecular surfaces in biological aggregates is a central concern in structural biochemistry. Self-assembly is also emerging as a new strategy in chemical synthesis, with the potential of generating non-biological structures having dimensions of 1-10(exp 2) nanometers. Structures in the upper part of this range of sizes are presently inaccessible through chemical synthesis, and the ability to prepare them would open a route to structures comparable in size (and perhaps complementary in function) to those that can be prepared by microlithography and other techniques of microfabrication.

  4. In-situ synthesis of nanoparticles via supersolubilizing micelle self-assembly

    Institute of Scientific and Technical Information of China (English)

    WANG DingCong


    In-situ synthesis of nano-particles using the self-assembly of molten salt and super soluble micellae was proposed based on a phenomenon of super solubilization of molten salt in reverse micellae and its self-assembly when the concentration reached up to 95% (w/w). The mechanism of the self-assembly indicates that the self-assembly of molten salt occurs in a reverse micelle where a homogenous phase is established between 5% (w/w) of a surfactant with a VB value of less than 1 and a hydrocarbon species. This synthesis has some unique features, such as being free of water, highly effective deposition and narrow distribution of particle size.

  5. In-situ synthesis of nanoparticles via supersolubilizing micelle self-assembly

    Institute of Scientific and Technical Information of China (English)


    In-situ synthesis of nano-particles using the self-assembly of molten salt and super soluble micellae was proposed based on a phenomenon of super solubilization of molten salt in reverse micellae and its self-assembly when the concentration reached up to 95%(w/w).The mechanism of the self-assembly indicates that the self-assembly of molten salt occurs in a reverse micelle where a homogenous phase is established between 5%(w/w)of a surfactant with a VB value of less than 1 and a hydrocarbon spe- cies.This synthesis has some unique features,such as being free of water,highly effective deposition and narrow distribution of particle size.

  6. Amphiphilic Janus gold nanoparticles prepared by interface-directed self-assembly: synthesis and self-assembly. (United States)

    Liu, Guannan; Tian, Jia; Zhang, Xu; Zhao, Hanying


    Materials with Janus structures are attractive for wide applications in materials science. Although extensive efforts in the synthesis of Janus particles have been reported, the synthesis of sub-10 nm Janus nanoparticles is still challenging. Herein, the synthesis of Janus gold nanoparticles (AuNPs) based on interface-directed self-assembly is reported. Polystyrene (PS) colloidal particles with AuNPs on the surface were prepared by interface-directed self-assembly, and the colloidal particles were used as templates for the synthesis of Janus AuNPs. To prepare colloidal particles, thiol-terminated polystyrene (PS-SH) was dissolved in toluene and citrate-stabilized AuNPs were dispersed in aqueous solution. Upon mixing the two solutions, PS-SH chains were grafted to the surface of AuNPs and amphiphilic AuNPs were formed at the liquid-liquid interface. PS colloidal particles decorated with AuNPs on the surfaces were prepared by adding the emulsion to excess methanol. On the surface, AuNPs were partially embedded in the colloidal particles. The outer regions of the AuNPs were exposed to the solution and were functionalized through the grafting of atom-transfer radical polymerization (ATRP) initiator. Poly[2-(dimethamino)ethyl methacrylate] (PDMAEMA) on AuNPs were prepared by surface-initiated ATRP. After centrifugation and dissolving the colloidal particles in tetrahydrofuran (THF), Janus AuNPs with PS and PDMAEMA on two hemispheres were obtained. In acidic pH, Janus AuNPs are amphiphilic and are able to emulsify oil droplets in water; in basic pH, the Janus AuNPs are hydrophobic. In mixtures of THF/methanol at a volume ratio of 1:5, the Janus AuNPs self-assemble into bilayer structures with collapsed PS in the interiors and solvated PDMAEMA at the exteriors of the structures.

  7. An autonomous molecular assembler for programmable chemical synthesis (United States)

    Meng, Wenjing; Muscat, Richard A.; McKee, Mireya L.; Milnes, Phillip J.; El-Sagheer, Afaf H.; Bath, Jonathan; Davis, Benjamin G.; Brown, Tom; O'Reilly, Rachel K.; Turberfield, Andrew J.


    Molecular machines that assemble polymers in a programmed sequence are fundamental to life. They are also an achievable goal of nanotechnology. Here, we report synthetic molecular machinery made from DNA that controls and records the formation of covalent bonds. We show that an autonomous cascade of DNA hybridization reactions can create oligomers, from building blocks linked by olefin or peptide bonds, with a sequence defined by a reconfigurable molecular program. The system can also be programmed to achieve combinatorial assembly. The sequence of assembly reactions and thus the structure of each oligomer synthesized is recorded in a DNA molecule, which enables this information to be recovered by PCR amplification followed by DNA sequencing.

  8. Total Chemical Synthesis,Assembly of Human Torque Teno Virus Genome

    Institute of Scientific and Technical Information of China (English)

    Zheng Hou; Gengfu Xiao


    Torque teno virus(TTV)is a nonenveloped virus containing a single-stranded,circular DNA genome of approximately 3.8kb.We completely synthesized the 3808 nucleotides of the TTV(SANBAN isolate)genome,which contains a hairpin structure and a GC-rich region.More than 100 overlapping oligonucleotides were chemically synthesized and assembled by polymerise chain assembly reaction(PCA),and the synthesis was completed with splicing by overlap extension(SOEing).This study establishes the methodological basis of the chemical synthesis of a viral genome for use as a live attenuated vaccine or gene therapy vector.

  9. TMEM14C is required for erythroid mitochondrial heme metabolism (United States)

    Yien, Yvette Y.; Robledo, Raymond F.; Schultz, Iman J.; Takahashi-Makise, Naoko; Gwynn, Babette; Bauer, Daniel E.; Dass, Abhishek; Yi, Gloria; Li, Liangtao; Hildick-Smith, Gordon J.; Cooney, Jeffrey D.; Pierce, Eric L.; Mohler, Kyla; Dailey, Tamara A.; Miyata, Non; Kingsley, Paul D.; Garone, Caterina; Hattangadi, Shilpa M.; Huang, Hui; Chen, Wen; Keenan, Ellen M.; Shah, Dhvanit I.; Schlaeger, Thorsten M.; DiMauro, Salvatore; Orkin, Stuart H.; Cantor, Alan B.; Palis, James; Koehler, Carla M.; Lodish, Harvey F.; Kaplan, Jerry; Ward, Diane M.; Dailey, Harry A.; Phillips, John D.; Peters, Luanne L.; Paw, Barry H.


    The transport and intracellular trafficking of heme biosynthesis intermediates are crucial for hemoglobin production, which is a critical process in developing red cells. Here, we profiled gene expression in terminally differentiating murine fetal liver-derived erythroid cells to identify regulators of heme metabolism. We determined that TMEM14C, an inner mitochondrial membrane protein that is enriched in vertebrate hematopoietic tissues, is essential for erythropoiesis and heme synthesis in vivo and in cultured erythroid cells. In mice, TMEM14C deficiency resulted in porphyrin accumulation in the fetal liver, erythroid maturation arrest, and embryonic lethality due to profound anemia. Protoporphyrin IX synthesis in TMEM14C-deficient erythroid cells was blocked, leading to an accumulation of porphyrin precursors. The heme synthesis defect in TMEM14C-deficient cells was ameliorated with a protoporphyrin IX analog, indicating that TMEM14C primarily functions in the terminal steps of the heme synthesis pathway. Together, our data demonstrate that TMEM14C facilitates the import of protoporphyrinogen IX into the mitochondrial matrix for heme synthesis and subsequent hemoglobin production. Furthermore, the identification of TMEM14C as a protoporphyrinogen IX importer provides a genetic tool for further exploring erythropoiesis and congenital anemias. PMID:25157825

  10. TMEM14C is required for erythroid mitochondrial heme metabolism. (United States)

    Yien, Yvette Y; Robledo, Raymond F; Schultz, Iman J; Takahashi-Makise, Naoko; Gwynn, Babette; Bauer, Daniel E; Dass, Abhishek; Yi, Gloria; Li, Liangtao; Hildick-Smith, Gordon J; Cooney, Jeffrey D; Pierce, Eric L; Mohler, Kyla; Dailey, Tamara A; Miyata, Non; Kingsley, Paul D; Garone, Caterina; Hattangadi, Shilpa M; Huang, Hui; Chen, Wen; Keenan, Ellen M; Shah, Dhvanit I; Schlaeger, Thorsten M; DiMauro, Salvatore; Orkin, Stuart H; Cantor, Alan B; Palis, James; Koehler, Carla M; Lodish, Harvey F; Kaplan, Jerry; Ward, Diane M; Dailey, Harry A; Phillips, John D; Peters, Luanne L; Paw, Barry H


    The transport and intracellular trafficking of heme biosynthesis intermediates are crucial for hemoglobin production, which is a critical process in developing red cells. Here, we profiled gene expression in terminally differentiating murine fetal liver-derived erythroid cells to identify regulators of heme metabolism. We determined that TMEM14C, an inner mitochondrial membrane protein that is enriched in vertebrate hematopoietic tissues, is essential for erythropoiesis and heme synthesis in vivo and in cultured erythroid cells. In mice, TMEM14C deficiency resulted in porphyrin accumulation in the fetal liver, erythroid maturation arrest, and embryonic lethality due to profound anemia. Protoporphyrin IX synthesis in TMEM14C-deficient erythroid cells was blocked, leading to an accumulation of porphyrin precursors. The heme synthesis defect in TMEM14C-deficient cells was ameliorated with a protoporphyrin IX analog, indicating that TMEM14C primarily functions in the terminal steps of the heme synthesis pathway. Together, our data demonstrate that TMEM14C facilitates the import of protoporphyrinogen IX into the mitochondrial matrix for heme synthesis and subsequent hemoglobin production. Furthermore, the identification of TMEM14C as a protoporphyrinogen IX importer provides a genetic tool for further exploring erythropoiesis and congenital anemias.

  11. Processing of heme and heme-containing proteins by bacteria. (United States)

    Stojiljkovic, Igor; Perkins-Balding, Donna


    An extensive amount of new knowledge on bacterial systems involved in heme processing has been accumulated in the last 10 years. We discuss common themes in heme transport across bacterial outer and inner membranes, emphasizing proteins and mechanisms involved. The processing of heme in the bacterial cytoplasm is extensively covered, and a new hypothesis about the fate of heme in the bacterial cell is presented. Auxiliary genes involved in heme utilization, i.e., TonB, proteases, proteins involved in heme storage and pigmentation, as well as genes involved in regulation of heme assimilation are reviewed.

  12. Synthesis and electrocatalytic water oxidation by electrode-bound helical peptide chromophore-catalyst assemblies. (United States)

    Ryan, Derek M; Coggins, Michael K; Concepcion, Javier J; Ashford, Dennis L; Fang, Zhen; Alibabaei, Leila; Ma, Da; Meyer, Thomas J; Waters, Marcey L


    Artificial photosynthesis based on dye-sensitized photoelectrosynthesis cells requires the assembly of a chromophore and catalyst in close proximity on the surface of a transparent, high band gap oxide semiconductor for integrated light absorption and catalysis. While there are a number of approaches to assemble mixtures of chromophores and catalysts on a surface for use in artificial photosynthesis based on dye-sensitized photoelectrosynthesis cells, the synthesis of discrete surface-bound chromophore-catalyst conjugates is a challenging task with few examples to date. Herein, a versatile synthetic approach and electrochemical characterization of a series of oligoproline-based light-harvesting chromophore-water-oxidation catalyst assemblies is described. This approach combines solid-phase peptide synthesis for systematic variation of the backbone, copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) as an orthogonal approach to install the chromophore, and assembly of the water-oxidation catalyst in the final step. Importantly, the catalyst was found to be incompatible with the conditions both for amide bond formation and for the CuAAC reaction. The modular nature of the synthesis with late-stage assembly of the catalyst allows for systematic variation in the spatial arrangement of light-harvesting chromophore and water-oxidation catalyst and the role of intrastrand distance on chromophore-catalyst assembly properties. Controlled potential electrolysis experiments verified that the surface-bound assemblies function as water-oxidation electrocatalysts, and electrochemical kinetics data demonstrate that the assemblies exhibit greater than 10-fold rate enhancements compared to the homogeneous catalyst alone.

  13. Synthesis, characterization and applications of ionic supramolecular assemblies (United States)

    Lin, Xinrong

    Supramolecular ionic assemblies not only provide alternatives to conventional polymers, but also introduce unique and interesting functions for the design of "smart" polymeric assemblies for use in a number of fields due to their programmable and reversible properties. Research in the area has led to an understanding of the connection between molecular contributions and macroscopic properties, as well as a range of applications from material processing/manufacuturing to energy transfer and storage. To this end, we have developed a library of charged building blocks based on ionic liquids to create functional supramolecular ionic assemblies. The polymeric ionic assemblies prepared from a di-phosphonium and poly (acrylic acid) were first studied and found to have the potential to be utilized as "smart" materials due to their ability to reversibly respond to stimuli such as temperature and pressure. With the interest of elucidating the molecular contributions to the bulk macroscopic material properties, six supramolecular assemblies were sequentially characterized in terms of thermal, rheological and X-ray studies. The effect of side alkyl chain was found to dramatically change the material properties. A second type of supramolecular assembly was investigated based on a poly-phosphonium ionic liquid, which was complexed with a number of carboxylic acids. The material properties were easily manipulated from a sticky fiber to a brittle solid by changing the composition of the carboxylic acid. A crosslinked supramolecular assembly combining ionic interactions and weak covalent bonds, specifically disulfide bonds, was next designed and characterized. The network properties could be switched between "on and off" using mild conditions. The polymeric ionic networks and their building block ionic liquids are also of interest as safe electrolytes in energy storage devices due to their non-flammability, non-volatility, etc. We have identified one ionic liquid with superior

  14. Design, Synthesis, Assembly, and Engineering of Peptoid Nanosheets. (United States)

    Robertson, Ellen J; Battigelli, Alessia; Proulx, Caroline; Mannige, Ranjan V; Haxton, Thomas K; Yun, Lisa; Whitelam, Stephen; Zuckermann, Ronald N


    Two-dimensional (2D) atomically defined organic nanomaterials are an important material class with broad applications. However, few general synthetic methods exist to produce such materials in high yields and to precisely functionalize them. One strategy to form ordered 2D organic nanomaterials is through the supramolecular assembly of sequence-defined synthetic polymers. Peptoids, one such class of polymer, are designable bioinspired heteropolymers whose main-chain length and monomer sequence can be precisely controlled. We have recently discovered that individual peptoid polymers with a simple sequence of alternating hydrophobic and ionic monomers can self-assemble into highly ordered, free-floating nanosheets. A detailed understanding of their molecular structure and supramolecular assembly dynamics provides a robust platform for the discovery of new classes of nanosheets with tunable properties and novel applications. In this Account, we discuss the discovery, characterization, assembly, molecular modeling, and functionalization of peptoid nanosheets. The fundamental properties of peptoid nanosheets, their mechanism of formation, and their application as robust scaffolds for molecular recognition and as templates for the growth of inorganic minerals have been probed by an arsenal of experimental characterization techniques (e.g., scanning probe, electron, and optical microscopy, X-ray diffraction, surface-selective vibrational spectroscopy, and surface tensiometry) and computational techniques (coarse-grained and atomistic modeling). Peptoid nanosheets are supramolecular assemblies of 16-42-mer chains that form molecular bilayers. They span tens of microns in lateral dimensions and freely float in water. Their component chains are highly ordered, with chains nearly fully extended and packed parallel to one another as a result of hydrophobic and electrostatic interactions. Nanosheets form via a novel interface-catalyzed monolayer collapse mechanism. Peptoid

  15. Synthesis and self-assembly of glycal-based bolaforms. (United States)

    Bozell, Joseph J; Tice, Nathan C; Sanyal, Nibedita; Thompson, David; Kim, Jong-Mok; Vidal, Sébastien


    Glycal-based bolaforms serve as synthetically flexible components of molecular self-assembly. The compounds are prepared in good yield by a Ferrier reaction between triacetylglucal or -galactal or diacetylxylal and a long chain alpha,omega-diol, followed by deacetylation under Zemplen conditions. The reactions are stereoselective and preferentially afford the alpha-diastereomer. The bolaforms undergo self-assembly in water or water/dioxane solution to give a variety of nanostructures. In solution, bolaforms with C8 or C10 chains between glucal headgroups form nanoscale vesicles. In contrast, bolaforms with C12 chains exhibit lower solubility and a dynamic self-assembly, forming several different nanoscale structures. However, the solid-state structures of C12 bolaform isomers adopt shapes very similar to those of bolaforms possessing more extensive hydrogen-bonding networks, indicating that multiple hydrogen bonds in solution are important to formation of stable, discrete nanostructures but that only a few key intermolecular interactions between bolaform headgroups are necessary to determine the structure in the solid state. The diversity and differentiation of the functional groups present in glycal-based bolaforms suggest that they could be useful probes of the various noncovalent forces controlling the structure of new nanomaterials.

  16. Bottlebrush Polymers: Synthesis, Rheology, and Self-Assembly (United States)

    Dalsin, Samuel J.

    Bottlebrush polymers are comb-like molecules with a high density of side chains grafted along a central backbone. Due to their unique conformational properties, bottlebrush polymers have become attractive candidates for developing new photonic bandgap materials, nanotubes and nanowires, or drug delivery vehicles, to name a few. This dissertation primarily investigates the rheological properties and self-assembly behavior of bottlebrush polymer molecules made using a variety of different polymerization routes. A considerable portion of the work is directed towards the linear rheology of model, polyolefin-based bottlebrush polymers with independently varied branch and backbone lengths. These studies demonstrate how the tight spacing between branch points effectively precludes backbone entanglement in the polymer melts, but it does not inhibit the formation of entanglements among the branched side chains. Furthermore, the relaxation profiles reveal transient scaling behavior in which the dynamics transition from Zimm-like to Rouse-like at increasing relaxation times. These results highlight the distinct conformational character of bottlebrushes at different length scales. The latter parts of this work report on the self-assembly behavior of bottlebrush diblock polymers composed of atactic polypropylene and polystyrene side chains. The diblock samples are analyzed using small-angle X-ray scattering and atomic force microscopy. Nearly all of the samples display strong segregation between the two blocks, owing to the large molar mass of typical bottlebrush polymers. Consequently, only one experimental sample displays an accessible order-disorder transition temperature. The strong segregation is also shown to affect the ability of large bottlebrush diblocks to readily achieve well-ordered nanostructures by self-assembly. Finally, results of the most symmetric (by volume fraction) diblock samples are compared with predictions of a newly developed self-consistent field

  17. Synthesis and simultaneous self-assembly of novel antibacterial polyurethanes (United States)

    Duan, J. H.; Yin, F.; Jiang, G. C.


    Novel physically crosslinked polyurethane (PUII) based on isophorone diisocyanates (IPDI) was prepared by a conventional two step method. The chemical structures of the PUII were characterized by fourier transform infrared (FTIR), proton nuclear magnetic resonance (1H-NMR), gel permeation chromatography (GPC) and scanning electron microscopy (SEM). The PUII hydrogels were subjected to solvent-induced self-assembly in THF + water to construct a variety of morphologies. The self-assembly morphology of the PUII was observed by scanning electron microscopy (SEM). Different amounts (0.2%, 0.4%, 0.6%, 0.8%, 1.0%) of 1,3,5-Tris(2-hydroxyethyl)hexahydro-1,3,5-triazine (TNO) was added as antibacterial agent to the polyurethane prepolymers. The inhibiting capacity of the antibacterial films to the Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Gray mold has been studied. The inhibiting capacity of films for each strain effect became obvious with the increase of content of antibacterial agent and the sensitive degree to all kind of bacterial species was different.

  18. A self-assembled synthesis of carbon nanotubes for interconnects. (United States)

    Chen, Zexiang; Cao, Guichuan; Lin, Zulun; Koehler, Irmgard; Bachmann, Peter K


    We report a novel approach to grow highly oriented, freestanding and structured carbon nanotubes (CNTs) between two substrates, using microwave plasma chemical vapour deposition. Sandwiched, multi-layered catalyst structures are employed to generate such structures. The as-grown CNTs adhere well to both the substrate and the top contact, and provide a low-resistance electric contact between the two. High-resolution scanning electron microscope (SEM) images show that the CNTs grow perpendicular to these surfaces. This presents a simple way to grow CNTs in different, predetermined directions in a single growth step. The overall resistance of a CNT bundle and two CNT-terminal contacts is measured to be about 14.7 k Ω. The corresponding conductance is close to the quantum limit conductance G(0). This illustrates that our new approach is promising for the direct assembly of CNT-based interconnects in integrated circuits (ICs) or other micro-electronic devices.

  19. Dioxygen reactivity of meso-hydroxylated hemes: intermediates in heme degradation process catalyzed by heme oxygenase

    Indian Academy of Sciences (India)

    Sankar Prasad Rath


    Heme oxygenase (HO) is the only enzyme in mammals known to catalyse the physiological degradation of unwanted heme into biliverdin, Fe ion and CO. The process involves introduction of the hydroxyl group at one of its meso-positions as the first fundamental step of the heme cleavage process. It was also found that meso-amino heme undergoes similar ring-cleavage process while reacting with dioxygen in presence of pyridine as an axial ligand. The present paper briefly reviews the reactions of model meso-hydroxylated heme and its analogues with dioxygen, and their relevance in the heme degradation process.

  20. Triazolobithiophene Light Absorbing Self-Assembled Monolayers: Synthesis and Mass Spectrometry Applications

    Directory of Open Access Journals (Sweden)

    Denis Séraphin


    Full Text Available The synthesis of five light absorbing triazolobithiophenic thiols, which were utilized for producing self-assembled monolayers (SAMs on gold surfaces, is presented. The monolayer formation was monitored by cyclic voltammetry, indicating excellent surface coverage. The new triazolobithiophenic compounds exhibited an absorption maximum around 340 nm, which is close to the emission wavelength of a standard nitrogen laser. Consequently these compounds could be used to aid ionization in laser desorption mass spectrometry (MS.

  1. De novo synthesis and properties of analogues of the self-assembling chlorosomal bacteriochlorophylls

    Energy Technology Data Exchange (ETDEWEB)

    Mass, Olga [North Carolina State Univ., Raleigh, NC (United States); Pandithavidana, Dinesh R. [North Carolina State Univ., Raleigh, NC (United States); Ptaszek, Marcin [North Carolina State Univ., Raleigh, NC (United States); Santiago, Koraliz [North Carolina State Univ., Raleigh, NC (United States); Springer, Joseph W. [Washington Univ., St. Louis, MO (United States); Jiao, Jieying [Univ. Of California, Riverside, CA (United States); Tang, Qun [Univ. Of California, Riverside, CA (United States); Kirmaier, Christine [Washington Univ., St. Louis, MO (United States); Bocian, David F. [Univ. Of California, Riverside, CA (United States); Holten, Dewey [Washington Univ., St. Louis, MO (United States); Lindsey, Jonathan S. [North Carolina State Univ., Raleigh, NC (United States)


    Natural photosynthetic pigments bacteriochlorophyllsc, d and e in green bacteria undergo self-assembly to create an organized antenna system known as the chlorosome, which collects photons and funnels the resulting excitation energy toward the reaction centers. Mimicry of chlorosome function is a central problem in supramolecular chemistry and artificial photosynthesis, and may have relevance for the design of photosynthesis-inspired solar cells. The main challenge in preparing artificial chlorosomes remains the synthesis of the appropriate pigment (chlorin) equipped with a set of functional groups suitable to direct the assembly and assure efficient energy transfer. Prior approaches have entailed derivatization of porphyrins or semisynthesis beginning with chlorophylls. This paper reports a third approach, the de novo synthesis of macrocycles that contain the same hydrocarbon skeleton as chlorosomal bacteriochlorophylls. The synthesis here of Zn(II) 3-(1-hydroxyethyl)-10-aryl-13¹-oxophorbines (the aryl group consists of phenyl, mesityl, or pentafluorophenyl) entails selective bromination of a 3,13-diacetyl-10-arylchlorin, palladium-catalyzed 13¹-oxophorbine formation, and selective reduction of the 3-acetyl group using BH₃·tBuNH₂. Each macrocycle contains a geminal dimethyl group in the pyrroline ring to provide stability toward adventitious dehydrogenation. A Zn(II) 7-(1-hydroxyethyl)-10-phenyl-17-oxochlorin also has been prepared. Altogether, 30 new hydroporphyrins were synthesized. The UV-Vis absorption spectra of the new chlorosomal bacteriochlorophyll mimics reveal a bathochromic shift of [similar]1800 cm-1 of the Qy band in nonpolar solvent, indicating extensive assembly in solution. The Zn(II) 3-(1-hydroxyethyl)-10-aryl-13¹-oxophorbines differ in the propensity to form assemblies based on the 10-substituent in the following order: mesitylassemblies also can be formed in solid media and

  2. Clinically Important Features of Porphyrin and Heme Metabolism and the Porphyrias


    Siddesh Besur; Wehong Hou; Paul Schmeltzer; Bonkovsky, Herbert L.


    Heme, like chlorophyll, is a primordial molecule and is one of the fundamental pigments of life. Disorders of normal heme synthesis may cause human diseases, including certain anemias (X-linked sideroblastic anemias) and porphyrias. Porphyrias are classified as hepatic and erythropoietic porphyrias based on the organ system in which heme precursors (5-aminolevulinic acid (ALA), porphobilinogen and porphyrins) are chiefly overproduced. The hepatic porphyrias are further subdivided into acute p...

  3. Self assembled homodinuclear dithiocarbamates: One pot synthesis and spectral characterization (United States)

    Nami, Shahab A. A.; Husain, Ahmad; Ullah, Irfan


    Several self assembled homodinuclear complexes of the type [M2(Ldtc)2·4H2O] derived from quadridentate ligand (Ldtc), where Ldtc = 2-aminobenzoylhydrazidebis(dithiocarbamate) and M = Mn(II), Fe(II), Co(II), Ni(II), Cu(II) and Zn(II) have been reported. The in situ procedure gives high yield with the formation of single product as evident by TLC and various other physicochemical techniques. Elemental analysis, TGA, 1H NMR, 13C NMR, ESI mass spectrometry, EPR, UV-vis. and IR spectroscopy were used to characterize the homodinuclear complexes. The spectroscopic evidences and room temperature magnetic moment values suggest that all the complexes have octahedral geometry around the transition metal atom. A symmetrical bidentate coordination of the dithiocarbamato moiety has been observed in all the complexes. The energy-minimized structure of the molecule also showed that each metal atom acquires a distorted octahedral geometry. The complexes exhibit a three-step thermolytic pattern and are non-electrolyte in nature.

  4. Hydrothermal synthesis of self-assembled hierarchical tungsten oxides hollow spheres and their gas sensing properties. (United States)

    Li, Jinwei; Liu, Xin; Cui, Jiashan; Sun, Jianbo


    Hierarchical self-assembled hollow spheres (HS) of tungsten oxide nanosheets have been synthesized via a template-free hydrothermal method. Morphology evolution of the products is determined by the amount of H2C2O4 (oxalic acid) which serves as chelating agent. Structural features of the products were characterized by X-ray diffraction (XRD), and morphology was investigated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In addition, the porous structure was analyzed using the Brunauer-Emmett-Teller (BET) approach. The synthesis mechanism of the products with self-assembled hierarchical structures was proposed. The NO2 gas sensing properties of self-assembled hierarchical WO3 HS materials were investigated, the gas sensing properties of WO3 synthesized by a variety of formulations were compared, and the possible gas sensing mechanism was discussed. The obvious enhancement of the gas sensing properties was ascribed to the structure of the hierarchical HS.

  5. Synthesis of In2S3 nanoplates and their self-assembly into superlattices. (United States)

    Zhong, Haizheng; Ye, Mingfu; Zhou, Yi; Yang, Chunhe; Li, Yongfang


    Ultrathin nanoplates of beta-phase semiconductor In2S3 with the diameter of 50 - 80 nm and a thickness of 1.85 nm were synthesized via a simple hot injection solution method with Hexadecylamine (HDA) as ligand, and the nanoplates self-assembled into superlattices by face to face stacking during evaporation of the solvent. The nanoplates and the assembled superlattices were characterized by XRD, TEM, HRTEM, XPS etc. The TEM and XRD results display that the nanocrystals are highly crystalline, hexagonal shape, and uniform in thickness. A strong quantum confinement effect of the 2D nanoplates was observed from the UV-Vis and PL spectra for the first time in the In2S3 nanoplates. A possible mechanism of the synthesis and assembly was proposed.

  6. Assembly fabrication of linkers on glass surface and their effect on DNA synthesis and hybridization

    Institute of Scientific and Technical Information of China (English)

    ShenJiayao; XiaoPengfeng; HouPeng; JiMeiju; SunXiao; HeNongyue


    Linkers were assembled on a glass surface based on the hydrolysis and condensation of 3-glycidoxy-propyltrimethoxysilane (GPS). After the assembly of GPS, four approaches were tried to open the ending epoxide group of GPS or to further elongate the linkers. The effect of these approaches on DNA in situ synthesis and hybridization was investigated. For the spacing of the synthesis initiation sites, the wettability of the support and the length of the linking group that attaches the initiation site to the surface have direct influences on the yield of coupling reactions and the subsequent hybridization events. X-ray photoelectron spectroscopy (XPS) and mean contact angles of deionized water of the above slides were measured to assess the linker's characteristics in each procedure. It was proved that the glass slides were successfully modified and became excellent supports for the oligonucleotides synthesis. In addition, it proved best for the in situ oligonueleotides synthesis that a glass slide was in turn treated with ethylenediamine, glutaradehyde, ethanolamine and sodium borohydride solution at ambient temperature after silanized with GPS.

  7. Heme mediates cytotoxicity from artemisinin and serves as a general anti-proliferation target.

    Directory of Open Access Journals (Sweden)

    Shiming Zhang

    Full Text Available Heme (Fe2+ protoporphyrin IX is an essential molecule that has been implicated the potent antimalarial action of artemisinin and its derivatives, although the source and nature of the heme remain controversial. Artemisinins also exhibit selective cytotoxicity against cancer cells in vitro and in vivo. We demonstrate that intracellular heme is the physiologically relevant mediator of the cytotoxic effects of artemisinins. Increasing intracellular heme synthesis through the addition of aminolevulinic acid, protoporphyrin IX, or transferrin-bound iron increased the cytotoxicity of dihydroartemisinin, while decreasing heme synthesis through the addition of succinyl acetone decreased its cytotoxic activity. A simple and robust high throughput assay was developed to screen chemical compounds that were capable of interacting with heme. A natural products library was screened which identified the compound coralyne, in addition to artemisinin, as a heme interacting compound with heme synthesis dependent cytotoxic activity. These results indicate that cellular heme may serve a general target for the development of both anti-parasitic and anti-cancer therapeutics.

  8. Heme crystallization in the midgut of triatomine insects. (United States)

    Oliveira, Marcus F; Gandara, Ana Caroline P; Braga, Cláudia M S; Silva, José R; Mury, Flavia B; Dansa-Petretski, Marílvia; Menezes, Diego; Vannier-Santos, Marcos A; Oliveira, Pedro L


    Hemozoin (Hz) is a heme crystal produced by several blood-feeding organisms in order to detoxify free heme released upon hemoglobin (Hb) digestion. Here we show that heme crystallization also occurs in three species of triatomine insects. Ultraviolet-visible and infrared light absorption spectra of insoluble pigments isolated from the midgut of three triatomine species Triatoma infestans, Dipetalogaster maximus and Panstrongylus megistus indicated that all produce Hz. Morphological analysis of T. infestans and D. maximus midguts revealed the close association of Hz crystals to perimicrovillar membranes and also as multicrystalline assemblies, forming nearly spherical structures. Heme crystallization was promoted by isolated perimicrovillar membranes from all three species of triatomine bugs in vitro in heat-sensitive reactions. In conclusion, the data presented here indicate that Hz formation is an ancestral adaptation of Triatominae to a blood-sucking habit and that the presence of perimicrovillar membranes plays a central role in this process.

  9. Synthesis of self-assembly plasmonic silver nanoparticles with tunable luminescence color

    Energy Technology Data Exchange (ETDEWEB)

    Al-Ghamdi, Haifa S.; Mahmoud, Waleed E., E-mail:


    Assembly is an elegant and effective bottom-up approach to prepare arrays of nanoparticles from nobel metals. Noble metal nanoparticles are perfect building blocks because they can be prepared with an adequate functionalization to allow their assembly and with controlled sizes. Herein, we report a novel recipe for the synthesis of self-assembled silver nanoparticles with tunable optical properties and sizes. The synthetic route followed here based on the covalent binding among silver nanoparticles by means of poly vinyl alcohol for the first time. The size of silver nanoparticle is governed by varying the amount of sodium borohydride. The as-synthesized nanoparticles were characterized by transmission electron microscopy, x-ray diffraction, energy dispersive x-ray spectroscopy, selected area electron diffraction and UV–vis spectroscopy. Results depicted that self-assembly of mono-dispersed silver nanoparticles with different sizes have been achieved. The silver nanostructure has a single crystalline faced centered cubic structure with growth orientation along (1 1 1) facet. These nanoparticles exhibited localized surface plasmon resonance at 403 nm. The luminescence peaks were red-sifted from violet to green due to the increase of the particle sizes. -- Highlights: • Self-assembled silver nanoparticles based PVA were synthesized. • NaBH{sub 4} amount was found particle size dependent. • Silver nanoparticles strongly affected the surface plasmon resonance. • Highly symmetric luminescence emission band narrow width is obtained. • Dark field image showed a tunable color change from violet to green.

  10. Correlating self-assembly of block copolymers for their application in synthesis of gold nanoparticles. (United States)

    Ray, Debes; Aswall, Vinod Kumar; Srivastava, Dinesh


    We report the role of self-assembly of polyethylene oxide-polypropylene oxide-polyethylene oxide (PEO-PPO-PEO) block copolymers for the synthesis of gold nanoparticles from hydrogen tetrachloroaureate (III) hydrate (HAuCl4 x 3H2O) in aqueous solution. The synthesis has been carried out using three different block copolymers P85 [EO26PO39EO26], F88 [EO103PO39EO103] and P105 [EO37PO56EO37], which not only have varying molecular weight but also differ in hydrophobicity to hydrophilicity ratio. The formation of gold nanoparticles is confirmed by the UV-Visible Spectroscopy. Transmission electron microscopy (TEM) provides the sizes of the nanoparticles formed in these systems. Small-Angle Neutron Scattering (SANS) and Dynamic Light Scattering (DLS) techniques are used to correlate the self-assembly of block copolymer to their propensity to form gold nanoparticles. The yield is found to be in the order P105 > P85 > F88 and is related to the higher tendency of block copolymer to self-assemble to give greater yield of gold nanoparticles. For all the block copolymers, SANS and DLS results suggests that the yield in the synthesis does not always increases with the salt concentration and is limited due to the fact that most of the block copolymers remain unassociated with the gold nanoparticles. By making use of these unassociated block copolymers, we propose two methods (i) step addition method and (ii) additional reductant method, where the synthesis yield of gold nanoparticles can be enhanced by manifold.

  11. Precipitation synthesis and magnetic properties of self-assembled magnetite-chitosan nanostructures (United States)

    Bezdorozhev, Oleksii; Kolodiazhnyi, Taras; Vasylkiv, Oleg


    This paper reports the synthesis and magnetic properties of unique magnetite-chitosan nanostructures synthesized by the chemical precipitation of magnetite nanoparticles in the presence of chitosan. The influence of varying synthesis parameters on the morphology of the magnetic composites is determined. Depending on the synthesis parameters, magnetite-chitosan nanostructures of spherical (9-18 nm), rice-seed-like (75-290 nm) and lumpy (75-150 nm) shapes were obtained via self-assembly. Spherical nanostructures encapsulated by a 9-15 nm chitosan layer were assembled as well. The prospective morphology of the nanostructures is combined with their excellent magnetic characteristics. It was found that magnetite-chitosan nanostructures are ferromagnetic and pseudo-single domain. Rice-seed-like nanostructures exhibited a coercivity of 140 Oe and saturation magnetization of 56.7 emu/g at 300 K. However, a drop in the magnetic properties was observed for chitosan-coated spherical nanostructures due to the higher volume fraction of chitosan.

  12. Heme degrading protein HemS is involved in oxidative stress response of Bartonella henselae.

    Directory of Open Access Journals (Sweden)

    MaFeng Liu

    Full Text Available Bartonellae are hemotropic bacteria, agents of emerging zoonoses. These bacteria are heme auxotroph Alphaproteobacteria which must import heme for supporting their growth, as they cannot synthesize it. Therefore, Bartonella genome encodes for a complete heme uptake system allowing the transportation of this compound across the outer membrane, the periplasm and the inner membranes. Heme has been proposed to be used as an iron source for Bartonella since these bacteria do not synthesize a complete system required for iron Fe³⁺ uptake. Similarly to other bacteria which use heme as an iron source, Bartonellae must transport this compound into the cytoplasm and degrade it to allow the release of iron from the tetrapyrrole ring. For Bartonella, the gene cluster devoted to the synthesis of the complete heme uptake system also contains a gene encoding for a polypeptide that shares homologies with heme trafficking or degrading enzymes. Using complementation of an E. coli mutant strain impaired in heme degradation, we demonstrated that HemS from Bartonella henselae expressed in E. coli allows the release of iron from heme. Purified HemS from B. henselae binds heme and can degrade it in the presence of a suitable electron donor, ascorbate or NADPH-cytochrome P450 reductase. Knocking down the expression of HemS in B. henselae reduces its ability to face H₂O₂ induced oxidative stress.

  13. Facile synthesis of self-assembled biporous NiO and its electrochemical properties (United States)

    Muruganandham, M.; Suri, Rominder P. S.; Sillanpää, Mika; Lee, Gang-Juan; Wu, Jerry J.


    In this article, we report the synthesis of self-assembled bi-porous nickel oxide on a large scale without using any templates or matrix. Porous NiO microspheres composed of particles were obtained by thermal decomposition of nickel oxalate, which was prepared using nickel salt and oxalic acid as precursors. The as-obtained nickel oxalate and nickel oxide were characterized using X-ray powder diffraction (XRD), Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), X-ray Photoelectron Spectroscopy (XPS), thermogravimetric analysis (TGA), and nitrogen adsorption-desorption analysis. The influence of various experimental conditions on the formation nickel oxalate and NiO were studied. The nitrogen adsorption-desorption analysis showed that the synthesized NiO possesses a biporous (both mesoporous and macroporous) surface structur. The NiO microspheres showed a discharge capacity of 2929 mAh g-1. A plausible mechanism for the NiO self-assembly was proposed.

  14. Gene synthesis by integrated polymerase chain assembly and PCR amplification using a high-speed thermocycler (United States)

    TerMaat, Joel R.; Pienaar, Elsje; Whitney, Scott E.; Mamedov, Tarlan G.; Subramanian, Anuradha


    Polymerase chain assembly (PCA) is a technique used to synthesize genes ranging from a few hundred base pairs to many kilobase pairs in length. In traditional PCA, equimolar concentrations of single stranded DNA oligonucleotides are repeatedly hybridized and extended by a polymerase enzyme into longer dsDNA constructs, with relatively few full-length sequences being assembled. Thus, traditional PCA is followed by a second primer-mediated PCR reaction to amplify the desired full-length sequence to useful, detectable quantities. Integration of assembly and primer-mediated amplification steps into a single reaction using a high-speed thermocycler is shown to produce similar results. For the integrated technique, the effects of oligo concentration, primer concentration, and number of oligonucleotides are explored. The technique is successfully demonstrated for the synthesis of two genes encoding EPCR-1 (653 bp) and pUC19 β-lactamase (929 bp) in under 20 min. However, rapid integrated PCA–PCR was found to be problematic when attempted with the TM-1 gene (1509 bp). Partial oligonucleotide sets of TM-1 could be assembled and amplified simultaneously, indicating that the technique may be limited to a maximum number of oligonucleotides due to competitive annealing and competition for primers. PMID:19799938

  15. How Heme Oxygenase-1 Prevents Heme-Induced Cell Death.

    Directory of Open Access Journals (Sweden)

    Lilibeth Lanceta

    Full Text Available Earlier observations indicate that free heme is selectively toxic to cells lacking heme oxygenase-1 (HO-1 but how this enzyme prevents heme toxicity remains unexplained. Here, using A549 (human lung cancer and immortalized human bronchial epithelial cells incubated with exogenous heme, we find knock-down of HO-1 using siRNA does promote the accumulation of cell-associated heme and heme-induced cell death. However, it appears that the toxic effects of heme are exerted by "loose" (probably intralysosomal iron because cytotoxic effects of heme are lessened by pre-incubation of HO-1 deficient cells with desferrioxamine (which localizes preferentially in the lysosomal compartment. Desferrioxamine also decreases lysosomal rupture promoted by intracellularly generated hydrogen peroxide. Supporting the importance of endogenous oxidant production, both chemical and siRNA inhibition of catalase activity predisposes HO-1 deficient cells to heme-mediated killing. Importantly, it appears that HO-1 deficiency somehow blocks the induction of ferritin; control cells exposed to heme show ~10-fold increases in ferritin heavy chain expression whereas in heme-exposed HO-1 deficient cells ferritin expression is unchanged. Finally, overexpression of ferritin H chain in HO-1 deficient cells completely prevents heme-induced cytotoxicity. Although two other products of HO-1 activity--CO and bilirubin--have been invoked to explain HO-1-mediated cytoprotection, we conclude that, at least in this experimental system, HO-1 activity triggers the induction of ferritin and the latter is actually responsible for the cytoprotective effects of HO-1 activity.

  16. Serum iron increases with acute induction of hepatic heme oxygenase-1 in mice. (United States)

    Mostert, Volker; Nakayama, Akihiro; Austin, Lori M; Levander, Ximena A; Ferris, Christopher D; Hill, Kristina E; Burk, Raymond F


    Heme oxygenase (HO)-1 is induced by oxidative stress and protects against oxidant injury. We examined the effect of rapid induction of hepatic HO-1 on serum iron level. Serum iron was approximately doubled within 6 h when HO-1 was induced by phenobarbital treatment of selenium-deficient mice. Blocking heme synthesis with diethyl 1,4-dihydro-2,4,6-trimethyl-3,5-pyridinedicarboxylate (DDC) prevented the induction of HO-1 and the rise in serum iron. DDC did not block HO-1 induction by hemin. Inhibition of HO activity by tin protoporphyrin prevented a rise in serum iron that occurred following phorone treatment. These results indicate that heme synthesis or an exogenous source of heme is needed to allow induction of HO-1. Further, they link HO-1 induction with a rise in serum iron, suggesting that the iron resulting from catabolism of heme by HO-1 is released by the liver.

  17. Synthesis of Zeolites Using the ADOR (Assembly-Disassembly-Organization-Reassembly) Route. (United States)

    Wheatley, Paul S; Čejka, Jiří; Morris, Russell E


    Zeolites are an important class of materials that have wide ranging applications such as heterogeneous catalysts and adsorbents which are dependent on their framework topology. For new applications or improvements to existing ones, new zeolites with novel pore systems are desirable. We demonstrate a method for the synthesis of novel zeolites using the ADOR route. ADOR is an acronym for Assembly, Disassembly, Organization and Reassembly. This synthetic route takes advantage of the assembly of a relatively poorly stable that which can be selectively disassembled into a layered material. The resulting layered intermediate can then be organized in different manners by careful chemical manipulation and then reassembled into zeolites with new topologies. By carefully controlling the organization step of the synthetic pathway, new zeolites with never before seen topologies are capable of being synthesized. The structures of these new zeolites are confirmed using powder X-ray diffraction and further characterized by nitrogen adsorption and scanning electron microscopy. This new synthetic pathway for zeolites demonstrates its capability to produce novel frameworks that have never been prepared by traditional zeolite synthesis techniques.

  18. Molecular Design of Bioinspired Nanostructures for Biomedical Applications: Synthesis, Self-Assembly and Functional Properties (United States)

    Xu, Hesheng Victor; Zheng, Xin Ting; Mok, Beverly Yin Leng; Ibrahim, Salwa Ali; Yu, Yong; Tan, Yen Nee


    Biomolecules are the nanoscale building blocks of cells, which play multifaceted roles in the critical biological processes such as biomineralization in a living organism. In these processes, the biological molecules such as protein and nucleic acids use their exclusive biorecognition properties enabled from their unique chemical composition, shape and function to initiate a cascade of cellular events. The exceptional features of these biomolecules, coupled with the recent advancement in nanotechnology, have led to the emergence of a new research field that focuses on the molecular design of bioinspired nanostructures that inherit the extraordinary function of natural biomaterials. These “bioinspired” nanostructures could be formulated by biomimetic approaches through either self-assembling of biomolecules or acting as a biomolecular template/precursor to direct the synthesis of nanocomposite. In either situation, the resulting nanomaterials exhibit phenomenal biocompatibility, superb aqueous solubility and excellent colloidal stability, branding them exceptionally desirable for both in vitro and in vivo biomedical applications. In this review, we will present the recent developments in the preparation of “bioinspired” nanostructures through biomimetic self-assembly and biotemplating synthesis, as well as highlight their functional properties and potential applications in biomedical diagnostics and therapeutic delivery. Lastly, we will conclude this topic with some personal perspective on the challenges and future outlooks of the “bioinspired” nanostructures for nanomedicine.

  19. Biofabrication methods for the patterned assembly and synthesis of viral nanotemplates

    Energy Technology Data Exchange (ETDEWEB)

    Gerasopoulos, K; Ghodssi, R [MEMS Sensors and Actuators Laboratory (MSAL), University of Maryland, College Park, MD 20742 (United States); McCarthy, M [Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139 (United States); Banerjee, P [Department of Materials Science and Engineering, University of Maryland, College Park, MD 20742 (United States); Fan, X [Institute for Systems Research, University of Maryland, College Park, MD 20742 (United States); Culver, J N, E-mail: [Center for Biosystems Research, University of Maryland Biotechnology Institute, MD 20742 (United States)


    This paper reports on novel methodologies for the patterning and templated synthesis of virus-structured nanomaterials in two- and three-dimensional microfabricated architectures using the Tobacco mosaic virus (TMV). The TMV is a high aspect ratio biological molecule which can be engineered to include amino acids with enhanced binding properties. These modifications facilitate self-assembly of the TMV onto various substrates and enable its use as a template for the synthesis of nanostructured materials. This work focuses on the combination of this bottom-up biologically inspired fabrication method with standard top-down micromachining processes that allow direct integration of the virus-structured materials into batch-fabricated devices. Photolithographic patterning of uncoated as well as nickel-coated TMV nanostructures has been achieved using a lift-off process in both solvent and mild basic solutions and their assembly onto three-dimensional polymer and silicon microstructures is demonstrated. In addition to these patterning techniques, in situ formation of metal oxide TMV coatings in patterned microfabricated environments is shown using atomic layer deposition directly on the nickel-coated viruses. The biofabrication 'process toolbox' presented in this work offers a simple and versatile alternative for the hierarchical patterning and incorporation of biotemplated nanomaterials into micro/nanofabrication schemes.

  20. Copper(II)-Mediated Self-Assembly of Hairpin Peptides and Templated Synthesis of CuS Nanowires. (United States)

    Wang, Chengdong; Sun, Yawei; Wang, Jiqian; Xu, Hai; Lu, Jian R


    The self-assembly of peptides and proteins under well-controlled conditions underlies important nanostructuring processes that could be harnessed in practical applications. Herein, the synthesis of a new hairpin peptide containing four histidine residues is reported and the self-assembly process mediated by metal ions is explored. The work involves the combined use of circular dichroism, NMR spectroscopy, UV/Vis spectroscopy, AFM, and TEM to follow the structural and morphological details of the metal-coordination-mediated folding and self-assembly of the peptide. The results indicate that by forming a tetragonal coordination geometry with four histidine residues, copper(II) ions selectively trigger the peptide to fold and then self-assemble into nanofibrils. Furthermore, the copper(II)-bound nanofibrils template the synthesis of CuS nanowires, which display a near-infrared laser-induced thermal effect.

  1. The Legend of Sally Hemings (United States)

    Belz, Herman


    The part played by Sally Hemings in the life of Thomas Jefferson has been regarded as provocatively dubious since political enemy James Callender claimed in 1802 that Jefferson was the father of several of Hemings's children. Historian Merrill Peterson, observing that paternity is hard to prove, wrote in 1960 that no concrete evidence was ever…

  2. Synthesis of 5-hydroxyectoine from ectoine: crystal structure of the non-heme iron(II and 2-oxoglutarate-dependent dioxygenase EctD.

    Directory of Open Access Journals (Sweden)

    Klaus Reuter

    Full Text Available As a response to high osmolality, many microorganisms synthesize various types of compatible solutes. These organic osmolytes aid in offsetting the detrimental effects of low water activity on cell physiology. One of these compatible solutes is ectoine. A sub-group of the ectoine producer's enzymatically convert this tetrahydropyrimidine into a hydroxylated derivative, 5-hydroxyectoine. This compound also functions as an effective osmostress protectant and compatible solute but it possesses properties that differ in several aspects from those of ectoine. The enzyme responsible for ectoine hydroxylation (EctD is a member of the non-heme iron(II-containing and 2-oxoglutarate-dependent dioxygenases (EC 1.14.11. These enzymes couple the decarboxylation of 2-oxoglutarate with the formation of a high-energy ferryl-oxo intermediate to catalyze the oxidation of the bound organic substrate. We report here the crystal structure of the ectoine hydroxylase EctD from the moderate halophile Virgibacillus salexigens in complex with Fe(3+ at a resolution of 1.85 A. Like other non-heme iron(II and 2-oxoglutarate dependent dioxygenases, the core of the EctD structure consists of a double-stranded beta-helix forming the main portion of the active-site of the enzyme. The positioning of the iron ligand in the active-site of EctD is mediated by an evolutionarily conserved 2-His-1-carboxylate iron-binding motif. The side chains of the three residues forming this iron-binding site protrude into a deep cavity in the EctD structure that also harbours the 2-oxoglutarate co-substrate-binding site. Database searches revealed a widespread occurrence of EctD-type proteins in members of the Bacteria but only in a single representative of the Archaea, the marine crenarchaeon Nitrosopumilus maritimus. The EctD crystal structure reported here can serve as a template to guide further biochemical and structural studies of this biotechnologically interesting enzyme family.

  3. Synthesis of 5-hydroxyectoine from ectoine: crystal structure of the non-heme iron(II) and 2-oxoglutarate-dependent dioxygenase EctD. (United States)

    Reuter, Klaus; Pittelkow, Marco; Bursy, Jan; Heine, Andreas; Craan, Tobias; Bremer, Erhard


    As a response to high osmolality, many microorganisms synthesize various types of compatible solutes. These organic osmolytes aid in offsetting the detrimental effects of low water activity on cell physiology. One of these compatible solutes is ectoine. A sub-group of the ectoine producer's enzymatically convert this tetrahydropyrimidine into a hydroxylated derivative, 5-hydroxyectoine. This compound also functions as an effective osmostress protectant and compatible solute but it possesses properties that differ in several aspects from those of ectoine. The enzyme responsible for ectoine hydroxylation (EctD) is a member of the non-heme iron(II)-containing and 2-oxoglutarate-dependent dioxygenases (EC 1.14.11). These enzymes couple the decarboxylation of 2-oxoglutarate with the formation of a high-energy ferryl-oxo intermediate to catalyze the oxidation of the bound organic substrate. We report here the crystal structure of the ectoine hydroxylase EctD from the moderate halophile Virgibacillus salexigens in complex with Fe(3+) at a resolution of 1.85 A. Like other non-heme iron(II) and 2-oxoglutarate dependent dioxygenases, the core of the EctD structure consists of a double-stranded beta-helix forming the main portion of the active-site of the enzyme. The positioning of the iron ligand in the active-site of EctD is mediated by an evolutionarily conserved 2-His-1-carboxylate iron-binding motif. The side chains of the three residues forming this iron-binding site protrude into a deep cavity in the EctD structure that also harbours the 2-oxoglutarate co-substrate-binding site. Database searches revealed a widespread occurrence of EctD-type proteins in members of the Bacteria but only in a single representative of the Archaea, the marine crenarchaeon Nitrosopumilus maritimus. The EctD crystal structure reported here can serve as a template to guide further biochemical and structural studies of this biotechnologically interesting enzyme family.

  4. Effects of chronic exposure to sublethal concentrations of lead acetate on heme synthesis and immune function in red-tailed hawks. (United States)

    Redig, P T; Lawler, E M; Schwartz, S; Dunnette, J L; Stephenson, B; Duke, G E


    Red-tailed hawks were exposed to sublethal levels of lead acetate for periods of 3 or 11 weeks. Alterations in the heme biosynthetic pathway were demonstrated after the first week of exposure to 0.82 mg lead per kilogram body weight per day. Activity of erythrocyte porphobilinogen synthase (aminolevulinic acid dehydratase) was depressed significantly and did not return to normal levels until 5 weeks after the termination of lead treatments. A rapid and relatively brief increase in erythrocyte free protoporphyrin and a slower but more prolonged increase in its zinc complex were also demonstrated with exposure to this dose of lead for 3 weeks. Less substantial decreases in hematocrit and hemoglobin levels occurred but only in the longer experiment with exposure to higher lead levels. Short term, low level lead exposure did not effect immune function significantly in the hawks, as measured by antibody titers to foreign red blood cells or by the mitogenic stimulation of T-lymphocytes. Increased lead exposure produced a significant decrease in the mitogenic response but had no effect on antibody titers.

  5. Disk-shaped Symmetric Hexa-substituted Triphenylene Derivatives: Synthesis, Physical Properties and Self-assembly

    Institute of Scientific and Technical Information of China (English)

    WANG Dong; MI Yong-sheng; TANG Jian-kai; LIANG Peng-xia; JIN Zhao-kui; YANG Zhou; YANG Huai


    A series of triphenylene derivatives with six symmetric substituents was synthesized from hexabromotriphenylene.The synthesis was conducted by six-fold palladium-catalyzed Hagihara-Sonogashira crosscoupling reactions to yield the hexa-alkynyl substituted triphenylene derivatives of HTP1,HTP2,HTP3 and HTP4.The six symmetric substituents can not only endow the triphenylene the longer π-conjugated range,but also increase the solubility of the compounds.Their photophysical,electrochemical,thermal properties were investigated respectively.With the comparison of their properties,the structure-property relationships were established which demonstrated the influences of different substituents on the electronic nature and the mesomorphic phase of these disk-shaped molecules.In addition,with the scanning electron microscopy(SEM) and polarized optical microscopy(POM) characterization,the self-assembly behaviors of the compounds were also investigated.

  6. Synthesis and self-assembly of PAMAM/PAA Janus dendrimers (United States)

    Gao, Chunmei; Liu, Mingzhu; Lü, Shaoyu; Zhang, Xinjie; Chen, Yuanmou


    Janus dendrimers have two differently functionalized segments which are located on opposite sides. They have many excellent properties and broad application prospects. In this study, poly(amido amine)/poly(acrylic acid) (PAMAM/PAA) Janus dendrimers were prepared by click chemistry. One of the first steps taken was the synthesis of N-Boc-G3.0 PAMAM dendrimers with primary amine groups at the periphery. Second, by amide coupling between propargylic acid and N-Boc-G3.0 PAMAM, PAMAM dendrimers with alkyne were successfully synthesized. After being dissolved in aqueous solutions with different pH, Janus dendrimers spontaneously form flowerlike micellar, Janus particles, and spherical micelles due to primary amino, tertiary amino, and carboxyl groups in the dendrimers. This self-assembly behavior depending on pH changes has a number of potential applications in the field of materials.

  7. The heme-heme oxygenase system: a molecular switch in wound healing.

    NARCIS (Netherlands)

    Wagener, F.A.D.T.G.; Beurden, H.E. van; Hoff, J.W. Von den; Adema, G.J.; Figdor, C.G.


    When cells are injured they release their contents, resulting in a local accumulation of free heme proteins and heme. Here, we investigated the involvement of heme and its degrading enzyme heme oxygenase (HO) in the inflammatory process during wound healing. We observed that heme directly accumulate

  8. Heme in intestinal epithelial cell turnover, differentiation,detoxification, inflammation, carcinogenesis, absorption and motility

    Institute of Scientific and Technical Information of China (English)

    Phillip S Oates; Adrian R West


    The gastrointestinal tract is lined by a simple epithelium that undergoes constant renewal involving cell division,differentiation and cell death. In addition, the epithelial lining separates the hostile processes of digestion and absorption that occur in the intestinal lumen from the aseptic environment of the internal milieu by defensive mechanisms that protect the epithelium from being breached. Central to these defensive processes is the synthesis of heme and its catabolism by heme oxygenase (HO). Dietary heme is also an important source of iron for the body which is taken up intact by the enterocyte.This review describes the recent literature on the diverse properties of heme/HO in the intestine tract.The roles of heme/HO in the regulation of the cell cycle/apoptosis, detoxification of xenobiotics, oxidative stress,inflammation, development of colon cancer, hemeiron absorption and intestinal motility are specifically examined.

  9. Synthesis and Self-Assembly Behaviors of Polyhedral Oligomeric Silsesquioxane Based Giant Molecular Shape Amphiphiles (United States)

    Yue, Kan; Yu, Xinfei; Liu, Chang; Zhang, Wen-Bin; Cheng, Stephen


    Recently, our group has focus on the synthesis and characterization of novel giant molecular shape amphiphiles (GMSAs) based on functionalized molecular nanoparticles (MNPs), such as polyhedral oligomeric silsesquioxane (POSS), tethered with polymeric tails. A general synthetic method via the combination of sequential ?click? reactions has been developed and several model GMSAs with various tail lengths and distinct molecular topologies, which can be referred as the ?giant surfactants?, ?giant lipids?, ?giant gemini surfactants?, and ?giant bolaform surfactants? etc., have been demonstrated. Studies on their self-assembly behaviors in the bulk have revealed the formation of different ordered mesophase structures with feature sizes around 10 nanometers, which have been investigated in detail by small angle X-ray scattering (SAXS) technique and transmission electron microscopy (TEM). These findings have general implications on understanding the underlying principles of self-assembly behaviors of GMSAs, and might have potential applications in nano-patterning technology. This work is supported by NSF (DMR-0906898) and the Joint-Hope Foundation.

  10. Polymer-virus core-shell structures prepared via co-assembly and template synthesis methods

    Institute of Scientific and Technical Information of China (English)



    Bionanoparticles(BNPs),consisting of virus and virus-like assemblies,have attracted much attention in the biomedical field for their applications such as imaging and targeted drug delivery,owing to their well-defined structures and well-controlled chemistries.BNPs-based core-shell structures provide a unique system for the investigation of biological interactions such as protein-protein and protein-carbohydrate interactions.However,it is still a challenge to prepare the BNPs-based core-shell structures.Herein,we describe(i) co-assembly method and(ii) template synthesis method in the development of polymer-BNPs core-shell structures.These two methods can be divided into three different systems.In system A,different polymers including poly(2-vinylpyridine)(P2VP),poly(4-vinylpyridine)(P4VP) and poly(ε-caprolactone)-block-poly(2-vinylpyridine)(PCL-b-P2VP) can form a raspberry-like structure with BNPs.In system B,polystyrene(PS) spheres end capped with free amine and BNPs can form a core-shell structure.In System C,layer-by-layer(LBL) method is used to prepare positive charged PS particles,which can be used as a template to form the core-shell structures with BNPs.These two methods may open a new way for preparing novel protein-based functional materials for potential applications in the biomedical field.

  11. RAFT Synthesis and Self-Assembly of Free-Base Porphyrin Cored Star Polymers

    Directory of Open Access Journals (Sweden)

    Lin Wu


    Full Text Available Reversible addition fragmentation chain transfer (RAFT synthesis and self-assembly of free-base porphyrin cored star polymers are reported. The polymerization, in the presence of a free-base porphyrin cored chain transfer agent (CTA-FBP, produced porphyrin star polymers with controlled molecular weights and narrow polydispersities for a number of monomers including N, N-dimethylacrylamide (DMA and styrene (St. Well-defined amphiphilic star block copolymers, P-(PS-PDMA4 and P-(PDMA-PS4 (P: porphyrin, were also prepared and used for self-assembly studies. In methanol, a selective solvent for PDMA, spherical micelles were observed for both block copolymers as characterized by TEM. UV-vis studies suggested star-like micelles were formed from P-(PS-PDMA4, while P-(PDMA-PS4 aggregated into flower-like micelles. Spectrophotometric titrations indicated that the optical response of these two micelles to external ions was a function of micellar structures. These structure-related properties will be used for micelle studies and functional material development in the future.

  12. Synthesis and Characterization of Fatty Acid/Amino Acid Self-Assemblies

    Directory of Open Access Journals (Sweden)

    Joanna Gajowy


    Full Text Available In this paper, we discuss the synthesis and self-assembling behavior of new copolymers derived from fatty acid/amino acid components, namely dimers of linoleic acid (DLA and tyrosine derived diphenols containing alkyl ester pendent chains, designated as “R” (DTR. Specific pendent chains were ethyl (E and hexyl (H. These poly(aliphatic/aromatic-ester-amides were further reacted with poly(ethylene glycol (PEG and poly(ethylene glycol methyl ether of different molecular masses, thus resulting in ABA type (hydrophilic-hydrophobic-hydrophilic triblock copolymers. We used Fourier transform infrared (FTIR and nuclear magnetic resonance (NMR spectroscopies to evaluate the chemical structure of the final materials. The molecular masses were estimated by gel permeation chromatography (GPC measurements. The self-organization of these new polymeric systems into micellar/nanospheric structures in aqueous environment was evaluated using ultraviolet/visible (UV-VIS spectroscopy, dynamic light scattering (DLS and transmission electron microscopy (TEM. The polymers were found to spontaneously self-assemble into nanoparticles with sizes in the range 196–239 nm and critical micelle concentration (CMC of 0.125–0.250 mg/mL. The results are quite promising and these materials are capable of self-organizing into well-defined micelles/nanospheres encapsulating bioactive molecules, e.g., vitamins or antibacterial peptides for antibacterial coatings on medical devices.

  13. Templated Synthesis of Magnetic Nanoparticles through the Self-Assembly of Polymers and Surfactants

    Directory of Open Access Journals (Sweden)

    Vo Thu An Nguyen


    Full Text Available The synthesis of superparamagnetic nanoparticles (NPs for various technological applications continues to be an interesting research topic. The successful application of superparamagnetic NPs to each specific area typically depends on the achievement of high magnetization for the nanocrystals obtained, which is determined by their average size and size distribution. The size dispersity of magnetic NPs (MNPs is markedly improved when, during the synthesis, the nucleation and growth steps of the reaction are well-separated. Tuning the nucleation process with the assistance of a hosting medium that encapsulates the precursors (such as self-assembled micelles, dispersing them in discrete compartments, improves control over particle formation. These inorganic-organic hybrids inherit properties from both the organic and the inorganic materials, while the organic component can also bring a specific functionality to the particles or prevent their aggregation in water. The general concept of interest in this review is that the shape and size of the synthesized MNPs can be controlled to some extent by the geometry and the size of the organic templates used, which thus can be considered as molds at the nanometer scale, for both porous continuous matrices and suspensions.

  14. Synthesis and Characterization of Stimuli Responsive Block Copolymers, Self-Assembly Behavior and Applications

    Energy Technology Data Exchange (ETDEWEB)

    Determan, Michael Duane [Iowa State Univ., Ames, IA (United States)


    The central theme of this thesis work is to develop new block copolymer materials for biomedical applications. While there are many reports of stimuli-responsive amphiphilic [19-21] and crosslinked hydrogel materials [22], the development of an in situ gel forming, pH responsive pentablock copolymer is a novel contribution to the field, Figure 1.1 is a sketch of an ABCBA pentablock copolymer. The A blocks are cationic tertiary amine methacrylates blocked to a central Pluronic F127 triblock copolymer. In addition to the prerequisite synthetic and macromolecular characterization of these new materials, the self-assembled supramolecular structures formed by the pentablock were experimentally evaluated. This synthesis and characterization process serves to elucidate the important structure property relationships of these novel materials, The pH and temperature responsive behavior of the pentablock copolymer were explored especially with consideration towards injectable drug delivery applications. Future synthesis work will focus on enhancing and tuning the cell specific targeting of DNA/pentablock copolymer polyplexes. The specific goals of this research are: (1) Develop a synthetic route for gel forming pentablock block copolymers with pH and temperature sensitive properties. Synthesis of these novel copolymers is accomplished with ATRP, yielding low polydispersity and control of the block copolymer architecture. Well defined macromolecular characteristics are required to tailor the phase behavior of these materials. (2) Characterize relationship between the size and shape of pentablock copolymer micelles and gel structure and the pH and temperature of the copolymer solutions with SAXS, SANS and CryoTEM. (3) Evaluate the temperature and pH induced phase separation and macroscopic self-assembly phenomenon of the pentablock copolymer. (4) Utilize the knowledge gained from first three goals to design and formulate drug delivery formulations based on the multi

  15. Oegylated and cross-linking carbazole dendrons and dendrimers: Synthesis, characterization, assembly and thin film fabrication (United States)

    Felipe, Mary Jane Legaspi


    Dendrimers and dendrons (fractional dendrimers) are macromolecular structures that have well-defined molecular weights and precise number of functional groups. Tailoring these structures has provided designer molecules that can be used for various applications including drug delivery, sensors, and anti-biofouling surfaces. Overall, this dissertation provides novel protocols for the understanding of molecular design, synthesis, and structure-property relationship of OEGylated and conjugated carbazole dendrons and dendrimers. In this design, the use of oligo(ethylene glycol) (OEG) allows for the fabrication of biocompatible materials and imparts hydrophilicity on the structure while the carbazole functionality allows the cross-linking of these designer molecules. Such fine-tuning of macromolecular structures leading to the fabrication of anti-biofouling thin films, nanostructuring at the air-water interface, and assembly into supramolecular superstructures are considered in this dissertation. Chapter 2 details the synthesis, characterization, and electrochemical cross-linking of OEGylated linear dendrons and "Janus-type" dendrimers. Cross-linking the carbazole moieties enables the deposition of these films on Au, indium tin oxide-coated glass, and doped silicon through cyclic voltammetry and provides films with secondary level of organization imparted by the inter- and intra-molecular interaction among the carbazole units. Chapter 3 describes the fabrication of nonspecific protein adsorption resistant surfaces through electrochemical grafting of three different dendrons on SAM carbazole-coated gold substrates. The predictable shape of each dendron and the ability to cross-link the carbazole units have enabled parametrization of OEG conformation and density on these interfaces. Chapter 4 demonstrates the fundamental architectural requirements for obtaining stable films with OEGylated linear dendron molecules providing a new architectural design of nanostructuring

  16. Heme-iron complexing biphenyl and naphthalene derivatives as CYP17 inhibitorsfor the treatment of prostate cancer : design, synthesis and evaluation


    Pinto-Bazurco Mendieta, Mariano Aurelio Ernesto


    Insufficient efficacy and unsafe treatment options are the current state of the art of prostate cancer therapies. This motivated us to develop antiandrogenic drugs with reduced side effects, aiming at effective tumor growth prevention and regression for prostatic adenocarcinomas. The key enzyme in androgen synthesis CYP17 is to date the most promising target for an overall, mild treatment of prostate carcinomas. Thus, it was our aim to develop nonsteroidal CYP17 inhibitors. In the present wor...

  17. The P. aeruginosa Heme Binding Protein PhuS is a Heme Oxygenase Titratable Regulator of Heme Uptake



    The Pseudomonas aeruginosa heme utilization (Phu) system encodes several proteins involved in the acquisition of heme as an iron source. Once internalized heme is degraded by the iron-regulated heme oxygenase, HemO to biliverdin (BV) IXδ and β. In vitro studies have shown holo-PhuS transfers heme to the iron-regulated HemO. This protein-protein interaction is specific for HemO as PhuS does not interact with the α-regioselective heme oxygenase, BphO. Bacterial genetics and isotopic labeling...

  18. Heme on innate immunity and inflammation

    Directory of Open Access Journals (Sweden)

    Fabianno Ferreira Dutra


    Full Text Available Heme is an essential molecule expressed ubiquitously all through our tissues. Heme plays major functions in cellular physiology and metabolism as the prostetic group of diverse proteins. Once released from cells and from hemeproteins free heme causes oxidative damage and inflammation, thus acting as a prototypic damage-associated molecular pattern. In this context, free heme is a critical component of the pathological process of sterile and infectious hemolytic conditions including malaria, hemolytic anemias, ischemia-reperfusion and hemorrhage. The plasma scavanger proteins hemopexin and albumin reduce heme toxicity and are responsible for transporting free heme to intracellular compartments where it is catabolized by heme-oxygenase enzymes. Upon hemolysis or severe cellular damage the serum capacity to scavange heme may saturate and increase free heme to sufficient amounts to cause tissue damage in various organs. The mechanism by which heme causes reactive oxygen generation, activation of cells of the innate immune system and cell death are not fully understood. Although heme can directly promote lipid peroxidation by its iron atom, heme can also induce ROS generation and production of inflammatory mediators through the activation of selective signaling pathways. Heme activates innate immune cells such as macrophages and neutrophils through activation of innate immune receptors. The importance of these events has been demonstrated in infectious and non-infectious diseases models. In this review we will discuss the mechanisms behind heme-induced citotoxicity and inflammation and the consequences of these events on different tissues and diseases.

  19. Synchronous One-Pot (SOP) synthesis of hybrid structures: metal nanoparticles in self-assemblies of amphiphilic calix[6]biscrowns. (United States)

    Liang, Qing; Li, Changxi; Chen, Guosong; Jiang, Ming


    In this paper, we present a novel strategy, named Synchronous One-Pot (SOP) synthesis, to prepare gold nanoparticles (AuNPs) with a diameter of 2nm incorporated in self-assembled organic spheres with a diameter around 60nm (denoted as NPAs). Merits of this method include: (1) self-assembly of the organic component (calix[6]biscrown TAC) into spheres and the reduction of chloroauric acid (HAuCl(4)) take place simultaneously; (2) preparation combining UV irradiation and formaldehyde addition reduces the size and homogenizes the distribution of the resultant AuNPs within the TAC spheres. (3) Obtained material NPA gives attractive catalytic property to hydrogenation reaction.

  20. Structural Characterization of Heme Environmental Mutants of CgHmuT that Shuttles Heme Molecules to Heme Transporters

    Directory of Open Access Journals (Sweden)

    Norifumi Muraki


    Full Text Available Corynebacteria contain a heme uptake system encoded in hmuTUV genes, in which HmuT protein acts as a heme binding protein to transport heme to the cognate transporter HmuUV. The crystal structure of HmuT from Corynebacterium glutamicum (CgHmuT reveals that heme is accommodated in the central cleft with His141 and Tyr240 as the axial ligands and that Tyr240 forms a hydrogen bond with Arg242. In this work, the crystal structures of H141A, Y240A, and R242A mutants were determined to understand the role of these residues for the heme binding of CgHmuT. Overall and heme environmental structures of these mutants were similar to those of the wild type, suggesting that there is little conformational change in the heme-binding cleft during heme transport reaction with binding and the dissociation of heme. A loss of one axial ligand or the hydrogen bonding interaction with Tyr240 resulted in an increase in the redox potential of the heme for CgHmuT to be reduced by dithionite, though the wild type was not reduced under physiological conditions. These results suggest that the heme environmental structure stabilizes the ferric heme binding in CgHmuT, which will be responsible for efficient heme uptake under aerobic conditions where Corynebacteria grow.

  1. Giardia intestinalis incorporates heme into cytosolic cytochrome b₅. (United States)

    Pyrih, Jan; Harant, Karel; Martincová, Eva; Sutak, Robert; Lesuisse, Emmanuel; Hrdý, Ivan; Tachezy, Jan


    The anaerobic intestinal pathogen Giardia intestinalis does not possess enzymes for heme synthesis, and it also lacks the typical set of hemoproteins that are involved in mitochondrial respiration and cellular oxygen stress management. Nevertheless, G. intestinalis may require heme for the function of particular hemoproteins, such as cytochrome b5 (cytb5). We have analyzed the sequences of eukaryotic cytb5 proteins and identified three distinct cytb5 groups: group I, which consists of C-tail membrane-anchored cytb5 proteins; group II, which includes soluble cytb5 proteins; and group III, which comprises the fungal cytb5 proteins. The majority of eukaryotes possess both group I and II cytb5 proteins, whereas three Giardia paralogs belong to group II. We have identified a fourth Giardia cytb5 paralog (gCYTb5-IV) that is rather divergent and possesses an unusual 134-residue N-terminal extension. Recombinant Giardia cytb5 proteins, including gCYTb5-IV, were expressed in Escherichia coli and exhibited characteristic UV-visible spectra that corresponded to heme-loaded cytb5 proteins. The expression of the recombinant gCYTb5-IV in G. intestinalis resulted in the increased import of extracellular heme and its incorporation into the protein, whereas this effect was not observed when gCYTb5-IV containing a mutated heme-binding site was expressed. The electrons for Giardia cytb5 proteins may be provided by the NADPH-dependent Tah18-like oxidoreductase GiOR-1. Therefore, GiOR-1 and cytb5 may constitute a novel redox system in G. intestinalis. To our knowledge, G. intestinalis is the first anaerobic eukaryote in which the presence of heme has been directly demonstrated.

  2. Correction: A highly enantioselective Biginelli reaction using self-assembled methanoproline-thiourea organocatalysts: asymmetric synthesis of 6-isopropyl-3,4-dihydropyrimidines. (United States)

    Hang, Zhijun; Zhu, Jun; Lian, Xiang; Xu, Peng; Yu, Han; Han, Sheng


    Correction for 'A highly enantioselective Biginelli reaction using self-assembled methanoproline-thiourea organocatalysts: asymmetric synthesis of 6-isopropyl-3,4-dihydropyrimidines' by Zhijun Hang et al., Chem. Commun., 2016, 52, 80-83.

  3. Synthesis and characterization of DNA fenced, self-assembled SnO2 nano-assemblies for supercapacitor applications. (United States)

    Nithiyanantham, U; Ramadoss, Ananthakumar; Kundu, Subrata


    Self-assembled, aggregated, chain-like SnO2 nano-assemblies were synthesized at room temperature by a simple wet chemical route within an hour in the presence of DNA as a scaffold. The average size of the SnO2 particles and the chain diameter were controlled by tuning the DNA to Sn(ii) molar ratio and altering the other reaction parameters. A formation and growth mechanism of the SnO2 NPs on DNA is discussed. The SnO2 chain-like assemblies were utilized as potential anode materials in an electrochemical supercapacitor. From the supercapacitor study, it was found that the SnO2 nanomaterials showed different specific capacitance (Cs) values depending on varying chain-like morphologies and the order of Cs values was: chain-like (small size) > chain-like (large size). The highest Cs of 209 F g(-1) at a scan rate of 5 mV s(-1) was observed for SnO2 nano-assemblies having chain-like structure with a smaller size. The long term cycling stability study of a chain-like SnO2 electrode was found to be stable and retained ca. 71% of the initial specific capacitance, even after 5000 cycles. A supercapacitor study revealed that both morphologies can be used as a potential anode material and the best efficiency was observed for small sized chain-like morphology which is due to their higher BET surface area and specific structural orientation. The proposed route, by virtue of its simplicity and being environmentally benign, might become a future promising candidate for further processing, assembly, and practical application of other oxide based nanostructure materials.

  4. Solvothermal synthesis and controlled self-assembly of monodisperse titanium-based perovskite colloidal nanocrystals (United States)

    Caruntu, Daniela; Rostamzadeh, Taha; Costanzo, Tommaso; Salemizadeh Parizi, Saman; Caruntu, Gabriel


    The rational design of monodisperse ferroelectric nanocrystals with controlled size and shape and their organization into hierarchical structures has been a critical step for understanding the polar ordering in nanoscale ferroelectrics, as well as the design of nanocrystal-based functional materials which harness the properties of individual nanoparticles and the collective interactions between them. We report here on the synthesis and self-assembly of aggregate-free, single-crystalline titanium-based perovskite nanoparticles with controlled morphology and surface composition by using a simple, easily scalable and highly versatile colloidal route. Single-crystalline, non-aggregated BaTiO3 colloidal nanocrystals, used as a model system, have been prepared under solvothermal conditions at temperatures as low as 180 °C. The shape of the nanocrystals was tuned from spheroidal to cubic upon changing the polarity of the solvent, whereas their size was varied from 16 to 30 nm for spheres and 5 to 78 nm for cubes by changing the concentration of the precursors and the reaction time, respectively. The hydrophobic, oleic acid-passivated nanoparticles exhibit very good solubility in non-polar solvents and can be rendered dispersible in polar solvents by a simple process involving the oxidative cleavage of the double bond upon treating the nanopowders with the Lemieux-von Rudloff reagent. Lattice dynamic analysis indicated that regardless of their size, BaTiO3 nanocrystals present local disorder within the perovskite unit cell, associated with the existence of polar ordering. We also demonstrate for the first time that, in addition to being used for fabricating large area, crack-free, highly uniform films, BaTiO3 nanocubes can serve as building blocks for the design of 2D and 3D mesoscale structures, such as superlattices and superparticles. Interestingly, the type of superlattice structure (simple cubic or face centered cubic) appears to be determined by the type of solvent

  5. Synthesis, characterization and photocatalytic performance of self-assembled mesoporous TiO₂ nanoparticles. (United States)

    Lin, Yuan-Chung; Liu, Shou-Heng; Syu, Han-Ren; Ho, Tsung-Han


    A facile synthesis route is reported for preparation of mesoporous TiO(2) nanoparticles (MT-x) through evaporation induced self-assembly by using Pluronic F127, titanium isopropoxide, and various amounts of ethanol as templating agents, titanium sources and solvents, respectively. A variety of different spectroscopic and analytical techniques, such as small- and large-angle powder X-ray diffraction (XRD), N(2) adsorption-desorption isotherms, transmission electron microscopy (TEM), Raman and Fourier transform infrared (FTIR) spectroscopies were used to characterize the physicochemical properties of various MT-x catalysts. Among the catalysts, MT-20 was found to have better mesostructures formed by the arrangement of anatase TiO(2) nanoparticles of ca. 17.3 nm with broad interparticle pore size distribution. Hydrogen generation from water splitting on MT-20 using visible light was enhanced by at least 8.7 times if compared with the conventional TiO(2) photocatalyst. The superior photocatalytic performances observed for the synthesized MT-20 may be attributed to the presence of unique nanostructures in the TiO(2) photocatalysts.

  6. Synthesis and Self-Assembly of Amphiphilic Triblock Terpolymers with Complex Macromolecular Architecture

    KAUST Repository

    Polymeropoulos, George


    Two star triblock terpolymers (PS-b-P2VP-b-PEO)3 and one dendritic-like terpolymer [PS-b-P2VP-b-(PEO)2]3 of PS (polystyrene), P2VP (poly(2-vinylpyridine)), and PEO (poly(ethylene oxide)), never reported before, were synthesized by combining atom transfer radical and anionic polymerizations. The synthesis involves the transformation of the -Br groups of the previously reported Br-terminated 3-arm star diblock copolymers to one or two -OH groups, followed by anionic polymerization of ethylene oxide to afford the star or dendritic structure, respectively. The well-defined structure of the terpolymers was confirmed by static light scattering, size exclusion chromatography, and NMR spectroscopy. The self-assembly in solution and the morphology in bulk of the terpolymers, studied by dynamic light scattering and transmission electron microscopy, respectively, reveal new insights in the phase separation of these materials with complex macromolecular architecture. © 2015 American Chemical Society.

  7. Non-heme iron as ferrous sulfate does not interact with heme iron absorption in humans. (United States)

    Gaitán, Diego; Olivares, Manuel; Lönnerdal, Bo; Brito, Alex; Pizarro, Fernando


    The absorption of heme iron has been described as distinctly different from that of non-heme iron. Moreover, whether heme and non-heme iron compete for absorption has not been well established. Our objective was to investigate the potential competition between heme and non-heme iron as ferrous sulfate for absorption, when both iron forms are ingested on an empty stomach. Twenty-six healthy nonpregnant women were selected to participate in two iron absorption studies using iron radioactive tracers. We obtained the dose-response curve for absorption of 0.5, 10, 20, and 50 mg heme iron doses, as concentrated red blood cells. Then, we evaluated the absorption of the same doses, but additionally we added non-heme iron, as ferrous sulfate, at constant heme/non-heme iron molar ratio (1:1). Finally, we compare the two curves by a two-way ANOVA. Iron sources were administered on an empty stomach. One factor analysis showed that heme iron absorption was diminished just by increasing total heme iron (P heme iron as ferrous sulfate did not have any effect on heme iron absorption (P = NS). We reported evidence that heme and non-heme iron as ferrous sulfate does not compete for absorption. The mechanism behind the absorption of these iron sources is not clear.

  8. Fabrication of Zeolite A Rods with Irregular Macropores by Self-assembly of Zeolite A Microcrystals Using Microwave-assisted Hydrothermal Synthesis

    Institute of Scientific and Technical Information of China (English)

    程志林; 万惠霖; 刘赞


    Zeolite A rods by self-assembly of zeolite A microcrystal were successfully synthesized by microwave-assisted hydrothermal synthesis. The average size of zeolite crystals consisting of self-assembling materials was about 300 nm and the length of zeolite rods was in the range of 15-30 μm.

  9. Understanding the synthesis of mesoporous silica particles by evaporation induced self assembly (United States)

    Rathod, Shailendra B.


    Evaporation-induced self-assembly (EISA) of amphiphilic molecules within aerosol droplets is an attractive method for synthesis of mesoporous silica particles. The aim of this research was to demonstrate synthetic methodologies to develop novel particle architectures using this technique, and to understand the influence of the competing dynamics within an evaporating droplet undergoing EISA on the particle morphology and mesostructure. Experiments were conducted to control particle characteristics. Particle size and distribution was varied by varying the size and distribution of starting droplets. The compressed gas atomizer, TSI 3076, gave a roughly micron-sized droplets with a polydisperse population, whereas the vibrating orifice aerosol generator (VOAG), TSI 3450, gave a highly monodisperse droplet population when orifices of diameters 10 mum and 20 mum were used. The mesopore size and mesostructure ordering were varied by employing amphiphiles of different geometry and by the use of 1,2,3-trimethylbenzene, a pore-swelling agent. The extent of ordering was influenced by factors that govern the rates of reactions of the silica precursors relative to the rates of amphiphile self-assembly. These factors included acid concentration, the alkyl group in the tetraalkoxysilane precursor, the time for which the sol was aged before droplet generation, and CTAB/Si ratio in the starting sol. Experiments and simulation studies were carried out for particles made using CTAB as the templating agent and TMB as a pore-swelling agent. Analysis of these experiments was used to get insight into the three main dynamic processes occurring inside these droplets: evaporation of the volatile species, amphiphile self-assembly and phase transformation, and hydrolysis and condensation reactions of the silica precursor species. Pore swelling was observed for particles made using the VOAG. Particles made using the 10 mum orifice retained their hexagonal mesostructure upon addition of TMB in

  10. Synergistic self-assembly of scaffolds and building blocks for directed synthesis of organic nanomaterials

    Energy Technology Data Exchange (ETDEWEB)

    Dergunov, Sergey [ORNL; Richter, Andrew G [ORNL; Kim, Mariya D. [Saint Louis University; Pingali, Sai Venkatesh [ORNL; Urban, Volker S [ORNL; Pinkhassik, Eugene [University of Memphis


    Surfactants and hydrophobic monomers spontaneously assemble into vesicles containing monomers within the bilayer. The joint action of monomers and surfactants is essential in this synergistic self-assembly. Polymerization in the bilayer formed hollow polymer nanocapsules.

  11. pH-induced simultaneous synthesis and self-assembly of 3D layered beta-FeOOH nanorods. (United States)

    Fang, Xiao-Liang; Li, Yue; Chen, Cheng; Kuang, Qin; Gao, Xiang-Zhi; Xie, Zhao-Xiong; Xie, Su-Yuan; Huang, Rong-Bin; Zheng, Lan-Sun


    Higher-ordered architectures self-assembly of nanomaterials have recently attracted increasing attention. In this work, we report a spontaneous and efficient route to simultaneous synthesis and self-assembly of 3D layered beta-FeOOH nanorods depending on a pH-induced strategy, in which the continuous change of pH is achieved by hydrolysis of FeCl(3).6H(2)O in the presence of urea under hydrothermal conditions. The electron microscopy observations reveal that the square-prismic beta-FeOOH nanorods are self-assembled in a side-by-side fashion to form highly oriented 2D nanorod arrays, and the 2D nanorod arrays are further stacked in a face-to-face fashion to form the final 3D layered architectures. On the basis of time-dependent experiments, a multistage reaction mechanism for the formation of the 3D layered beta-FeOOH nanorods architecture is presented, involving the fast growth and synchronous self-assembly of the nanorods toward 1D, 2D, and 3D spontaneously. The experimental evidence further demonstrates that the urea-decomposition-dependent pH continuously changing in the solution, spontaneously altering the driving force competition between the electrostatic repulsive force and the attractive van der Waals force among the nanorods building blocks, is the essential factor to influence the self-assembly of the beta-FeOOH nanorods from 1D to 3D.

  12. Molecular Assemblies of Porphyrins and Macrocyclic Receptors: Recent Developments in Their Synthesis and Applications

    Directory of Open Access Journals (Sweden)

    Abdirahman A. Mohamod


    Full Text Available Metalloporphyrins which form the core of many bioenzymes and natural light harvesting or electron transport systems, exhibit a variety of selective functional properties depending on the state and surroundings with which they exist in biological systems. The specificity and ease with which they function in each of their bio-functions appear to be largely governed by the nature and disposition of the protein globule around the porphyrin reaction center. Synthetic porphyrin frameworks confined within or around a pre-organized molecular entity like the protein network in natural systems have attracted considerable attraction, especially in the field of biomimetic reactions. At the same time a large number of macrocyclic oligomers such as calixarenes, resorcinarenes, spherands, cyclodextrins and crown ethers have been investigated in detail as efficient molecular receptors. These molecular receptors are synthetic host molecules with enclosed interiors, which are designed three dimensionally to ensure strong and precise molecular encapsulation/recognition. Due to their complex structures, enclosed guest molecules reside in an environment isolated from the outside and as a consequence, physical properties and chemical reactions specific to that environment in these guest species can be identified. The facile incorporation of such molecular receptors into the highly photoactive and catalytically efficient porphyrin framework allows for convenient design of useful molecular systems with unique structural and functional properties. Such systems have provided over the years attractive model systems for the study of various biological and chemical processes, and the design of new materials and molecular devices. This review focuses on the recent developments in the synthesis of porphyrin assemblies associated with cyclodextrins, calixarenes and resorcinarenes and their potential applications in the fields of molecular encapsulation/recognition, and

  13. Allocation of Heme is Differentially Regulated by Ferrochelatase Isoforms in Arabidopsis Cells

    Directory of Open Access Journals (Sweden)

    Nino Asuela Espinas


    Full Text Available Heme is involved in various biological processes as a cofactor of hemoproteins located in various organelles. In plant cells, heme is synthesized by two isoforms of plastid-localized ferrochelatase, FC1 and FC2. In this study, by characterizing Arabidopsis T-DNA insertional mutants, we showed that the allocation of heme is differentially regulated by ferrochelatase isoforms in plant cells. Analyses of weak (fc1-1 and null (fc1-2 mutants suggest that FC1-producing heme is required for initial growth of seedling development. In contrast, weak (fc2-1 and null (fc2-2 mutants of FC2 showed pale green leaves and retarded growth, indicating that FC2-producing heme is necessary for chloroplast development. During the initial growth stage, FC2 deficiency caused reduction of plastid cytochromes. In addition, although FC2 deficiency marginally affected the assembly of photosynthetic reaction center complexes, it caused relatively larger but insufficient light-harvesting antenna to reaction centers, resulting in lower efficiency of photosynthesis. In the later vegetative growth, however, fc2-2 recovered photosynthetic growth, showing that FC1-producing heme may complement the FC2 deficiency. On the other hand, reduced level of cytochromes in microsomal fraction was discovered in fc1-1, suggesting that FC1-producing heme is mainly allocated to extraplastidic organelles. Furthermore, the expression of FC1 is induced by the treatment of an elicitor flg22 while that of FC2 was reduced, and fc1-1 abolished the flg22-dependent induction of FC1 expression and peroxidase activity. Consequently, our results clarified that FC2 produces heme for the photosynthetic machinery in the chloroplast, while FC1 is the housekeeping enzyme providing heme cofactor to the entire cell. In addition, FC1 can partly complement FC2 deficiency and is also involved in defense against stressful conditions.

  14. Molecular self-assembly: Design, synthesis, and characterization of peptidic materials for bio- and nano-technologies (United States)

    Lamm, Matthew S.

    The research presented in this dissertation focuses on the design, synthesis, and characterization of amphiphilic peptides capable of self-assembling into beta-sheet fibrils under specific aqueous solution conditions. The peptide design consists of two beta-sheet forming strands of alternating valine and lysine residues, flanking a central tetrapeptide sequence that contains a diproline. Depending on the chirality of the prolines the peptide can assume either an intramolecularly folded or an extended conformation in the self-assembled state. For the peptide where intramolecular folding is designed against, the self-assembled nanostructure was found to exhibit a unique, nontwisted laminated morphology. Experimental techniques including transmission electron microscopy, atomic force microscopy, circular dichroism spectroscopy, Fourier transform infra-red spectroscopy and x-ray diffraction were employed to characterize the self-assembled structure and kinetics. With the understanding of the self-assembly process gained from these first peptides, other peptide sequences were rationally designed to assemble with a desired nanostructure. For example, the effect of peptide strand length in conserving the laminated morphology and controlling the fibril height was investigated. In addition, other peptides were designed so as to affect the self-assembled nanostructure by enforcing a parallel versus anti-parallel beta-sheet or by disrupting the registry of laminating beta-sheet filaments. In some cases, the peptides assembled into structures predicted by the initial design while other peptides assembled into unexpected fibril morphologies. The major conclusions from the research on these peptides is that the diproline sequence plays an important role in disrupting the beta-strand twist thereby resulting in a nontwisted laminated morphology. However, when the flanking beta-strands become long enough, the effect of the diproline sequence becomes diminished and the fibrils do

  15. Insitu synthesis of self-assembled gold nanoparticles on glass or silicon substrates through reactive inkjet printing

    KAUST Repository

    Abulikemu, Mutalifu


    A facile and low cost method for the synthesis of self-assembled nanoparticles (NPs) with minimal size variation and chemical waste by using reactive inkjet printing was developed. Gold NPs with diameters as small as (8±2)nm can be made at low temperature (120 °C). The size of the resulting NPs can be readily controlled through the concentration of the gold precursor and oleylamine ink. The pure gold composition of the synthesized NPs was confirmed by energy-dispersive X-ray spectroscopy (EDXS) analysis. High-resolution SEM (HRSEM) and TEM (HRTEM), and X-ray diffraction revealed their size and face-centered cubic (fcc) crystal structure, respectively. Owing to the high density of the NP film, UV/Vis spectroscopy showed a red shift in the intrinsic plasmonic resonance peak. We envision the extension of this approach to the synthesis of other nanomaterials and the production of tailored functional nanomaterials and devices. Midas touch: The use of low-cost manufacturing approaches in the synthesis of nanoparticles is critical for many applications. Reactive inkjet printing, along with a judicious choice of precursor/solvent system, was used to synthesize a relatively uniform assembly of crystalline gold nanoparticles, with diameters as small as (8±2)nm, over a given substrate surface. © 2014 WILEY-VCH Verlag GmbH.

  16. Structural and Functional Models of Non-Heme Iron Enzymes : A Study of the 2-His-1-Carboxylate Facial Triad Structural Motif

    NARCIS (Netherlands)

    Bruijnincx, P.C.A.


    The structural and functional modeling of a specific group of non-heme iron enzymes by the synthesis of small synthetic analogues is the topic of this thesis. The group of non-heme iron enzymes with the 2-His-1-carboxylate facial triad has recently been established as a common platform for the activ

  17. Cell-free synthesis of the H-cluster: a model for the in vitro assembly of metalloprotein metal centers. (United States)

    Kuchenreuther, Jon M; Shiigi, Stacey A; Swartz, James R


    Many organometallic cofactors are highly complex and require multiple accessory proteins for both their assembly and transfer to a target protein. A cell-free system in which the biosynthetic pathway for a prosthetic group has been fully or even partially reconstructed enables investigations of the reaction sequence as well as the cofactor itself. As a model for the in vitro assembly of protein-bound metal centers, we describe a procedure for the cell-free synthesis of the H-cluster in the context of producing purified and active [FeFe] hydrogenase samples for spectroscopic studies. In general terms, this in vitro system is a combination of non-purified accessory proteins, exogenous substrates, and purified hydrogenase apoprotein. We also describe methods for making the required components used in the cell-free system. Specifically, these procedures include anaerobic expression of heterologous metalloproteins in Escherichia coli, anaerobic cell lysate production, and anaerobic metalloprotein purification using Strep-Tactin(®) chromatography.

  18. Synthesis of Nitrogen-Doped Mesoporous Carbon Spheres with Extra-Large Pores through Assembly of Diblock Copolymer Micelles

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Jing [Waseda University, Tokyo, Japan; Liu, Jiang [Curtin University of Technology, Perth, Australia; Li, Cuiling [KEK, Tsukuba, Ibaraki, Japan; Li, Yunqi [Waseda University, Tokyo, Japan; Tade, Moses O. [Curtin University of Technology, Perth, Australia; Dai, Sheng [ORNL; Yamauchi, Yusuke [Waseda University, Tokyo, Japan


    In this study, the synthesis of highly nitrogen-doped mesoporous carbon spheres (NMCS) is reported. The large pores of the NMCS were obtained through self-polymerization of dopamine (DA) and spontaneous co-assembly of diblock copolymer micelles. The resultant narrowly dispersed NMCS possess large mesopores (ca. 16 nm) and small particle sizes (ca. 200 nm). Lastly, the large pores and small dimensions of the N-heteroatom-doped carbon spheres contribute to the mass transportation by reducing and smoothing the diffusion pathways, leading to high electrocatalytic activity.

  19. DNA-surfactant complexes : preparation, self-assembly properties and applications in synthesis and bioelectronics

    NARCIS (Netherlands)

    Liu, Kai


    The powerful ionic self-assembly behavior of DNA-surfactant complexes make it a unique material for various applications from optoelectronics to biomedicine. Three types of DNA-surfactant assemblies, including bulk films, lyotropic liquid crystals (LCs) and hydrogels have been investigated extensive

  20. Synthesis, characterization and self-assembly of Co3+ complexes appended with phenol and catechol groups

    Indian Academy of Sciences (India)

    Afsar Ali; Deepak Bansal; Rajeev Gupt


    This work presents the syntheses, characterization and hydrogen bonding based self-assembly of Co3+ complexes of pyridine-amide based bidentate ligands containing appended phenol and catechol groups. Placement of multiple hydrogen bond donors (phenolic OH and amidic NH groups) and acceptors (Oamide groups) in these molecules results in interesting self-assembled architectures.

  1. Single-step microfluidic synthesis of various nonspherical polymer nanoparticles via in situ assembling: dominating role of polyelectrolytes molecules. (United States)

    Visaveliya, Nikunjkumar; Köhler, J Michael


    In this paper, a microfluidic approach has been used for the synthesis of ellipsoidal, dumbbell, rodlike, and necklacelike polymer nanoparticles. High yields of special types of nonspherical nanoparticles have been achieved by the implementation of an emulsion polymerization into microfluidic arrangement with a micro hole-plate reactor for the formation of monomer droplets. Here, in particular, the formation of nonspherical polymer nanoparticles is dependent on the presence of polyelectrolyte surface active molecules such as poly(4-styrenesulfonic acid-co-maleic acid) sodium salt (PSS-co-PM), poly(sodium-p-styrenesulfonate) (PSSS), and polyanetholesulfonic acid sodium salt (PAES). The shapes and sizes of the interparticle nanoassemblies are precisely controlled by adjusting the concentration of polyelectrolytes in the aqueous phase, and by choosing suitable flow rate ratios (aqueous to monomer phase), respectively. The formation of polymer nanoparticles with different morphologies can be explained by a spontaneous in situ assembling under partial electrostatic repulsive control in the single step synthesis. The effect of particle charge and the competition between thermal motion of particles and electrostatic repulsion on the spontaneous assembling under the condition of a limited polarizability are discussed here as an important factor for the formation process of nonspherical polymer nanoparticles.

  2. Heme Oxygenase-1 and Breast Cancer Resistance Protein Protect Against Heme-induced Toxicity

    NARCIS (Netherlands)

    Wagener, F.A.D.T.G.; Dankers, A.C.A.; Summeren, F. van; Scharstuhl, A.; Heuvel, J.J. van den; Koenderink, J.B.; Pennings, S.W.C.; Russel, F.G.M.; Masereeuw, R.


    Heme is the functional group of diverse hemoproteins and crucial for many cellular processes. However, heme is increasingly recognized as a culprit for a wide variety of pathologies, including sepsis, malaria, and kidney failure. Excess of free heme can be detrimental to tissues by mediating oxidati

  3. Rapamycin Induces Heme Oxygenase-1 in Liver but Inhibits Bile Flow Recovery after Ischemia

    NARCIS (Netherlands)

    Kist, Alwine; Wakkie, Joris; Madu, Max; Versteeg, Ruth; ten Berge, Judith; Nikolic, Andrej; Nieuwenhuijs, Vincent B.; Porte, Robert J.; Padbury, Robert T. A.; Barritt, Greg J.


    Background/Aims. Rapamycin, which is employed in the management of patients undergoing liver surgery, induces the synthesis of heme oxygenase-1 (HO-1) in some non-liver cell types. The aim was to investigate whether rapamycin can induce HO-1 expression in the liver, and to test the effects of rapamy

  4. Clinically Important Features of Porphyrin and Heme Metabolism and the Porphyrias (United States)

    Besur, Siddesh; Hou, Weihong; Schmeltzer, Paul; Bonkovsky, Herbert L.


    Heme, like chlorophyll, is a primordial molecule and is one of the fundamental pigments of life. Disorders of normal heme synthesis may cause human diseases, including certain anemias (X-linked sideroblastic anemias) and porphyrias. Porphyrias are classified as hepatic and erythropoietic porphyrias based on the organ system in which heme precursors (5-aminolevulinic acid (ALA), porphobilinogen and porphyrins) are chiefly overproduced. The hepatic porphyrias are further subdivided into acute porphyrias and chronic hepatic porphyrias. The acute porphyrias include acute intermittent, hereditary copro-, variegate and ALA dehydratase deficiency porphyria. Chronic hepatic porphyrias include porphyria cutanea tarda and hepatoerythropoietic porphyria. The erythropoietic porphyrias include congenital erythropoietic porphyria (Gűnther’s disease) and erythropoietic protoporphyria. In this review, we summarize the key features of normal heme synthesis and its differing regulation in liver versus bone marrow. In both organs, principal regulation is exerted at the level of the first and rate-controlling enzyme, but by different molecules (heme in the liver and iron in the bone marrow). We also describe salient clinical, laboratory and genetic features of the eight types of porphyria. PMID:25372274

  5. Clinically important features of porphyrin and heme metabolism and the porphyrias. (United States)

    Besur, Siddesh; Hou, Wehong; Schmeltzer, Paul; Bonkovsky, Herbert L


    Heme, like chlorophyll, is a primordial molecule and is one of the fundamental pigments of life. Disorders of normal heme synthesis may cause human diseases, including certain anemias (X-linked sideroblastic anemias) and porphyrias. Porphyrias are classified as hepatic and erythropoietic porphyrias based on the organ system in which heme precursors (5-aminolevulinic acid (ALA), porphobilinogen and porphyrins) are chiefly overproduced. The hepatic porphyrias are further subdivided into acute porphyrias and chronic hepatic porphyrias. The acute porphyrias include acute intermittent, hereditary copro-, variegate and ALA dehydratase deficiency porphyria. Chronic hepatic porphyrias include porphyria cutanea tarda and hepatoerythropoietic porphyria. The erythropoietic porphyrias include congenital erythropoietic porphyria (Gűnther's disease) and erythropoietic protoporphyria. In this review, we summarize the key features of normal heme synthesis and its differing regulation in liver versus bone marrow. In both organs, principal regulation is exerted at the level of the first and rate-controlling enzyme, but by different molecules (heme in the liver and iron in the bone marrow). We also describe salient clinical, laboratory and genetic features of the eight types of porphyria.

  6. Clinically Important Features of Porphyrin and Heme Metabolism and the Porphyrias

    Directory of Open Access Journals (Sweden)

    Siddesh Besur


    Full Text Available Heme, like chlorophyll, is a primordial molecule and is one of the fundamental pigments of life. Disorders of normal heme synthesis may cause human diseases, including certain anemias (X-linked sideroblastic anemias and porphyrias. Porphyrias are classified as hepatic and erythropoietic porphyrias based on the organ system in which heme precursors (5-aminolevulinic acid (ALA, porphobilinogen and porphyrins are chiefly overproduced. The hepatic porphyrias are further subdivided into acute porphyrias and chronic hepatic porphyrias. The acute porphyrias include acute intermittent, hereditary copro-, variegate and ALA dehydratase deficiency porphyria. Chronic hepatic porphyrias include porphyria cutanea tarda and hepatoerythropoietic porphyria. The erythropoietic porphyrias include congenital erythropoietic porphyria (Gűnther’s disease and erythropoietic protoporphyria. In this review, we summarize the key features of normal heme synthesis and its differing regulation in liver versus bone marrow. In both organs, principal regulation is exerted at the level of the first and rate-controlling enzyme, but by different molecules (heme in the liver and iron in the bone marrow. We also describe salient clinical, laboratory and genetic features of the eight types of porphyria.

  7. Mechanisms of assembly of the enzyme-ssDNA complexes required for recombination-dependent DNA synthesis and repair in bacteriophage T4

    Energy Technology Data Exchange (ETDEWEB)

    Morrical, S.; Hempstead, K.; Morrical, M. [Univ. of Vermont College of Medicine, Burlington, VT (United States)


    During late stages of bacteriophage T4 infection in E. coli, the initiation of phage DNA replication is dependent on the homologous recombination activity of the T4 uvsX protein. In vitro, uvsX protein initiates DNA synthesis on a duplex template by inserting the 3{prime} end of a homologous ssDNA molecule into the duplex. The resulting D-loop structure serves as a primer-template junction for the assembly of the T4 replication fork. Two key steps in this initiation process are (A) the assembly of uvsX-ssDNA complexes necessary for recombination activity and for the priming of lead-strand DNA synthesis, and (B) the assembly of the T4 primosome (gp41 helicase/gp61 primase complex) onto the single-stranded template for lagging-strand synthesis. Our laboratory is focusing on the mechanisms of these two different but related enzyme-ssDNA assembly processes. In this extended abstract, we describe recent efforts in our laboratory to elucidate the mechanism by which the gp41 helicase enzyme is assembled onto gp32-covered ssDNA, a process requiring the activity of a special helicase assembly factor, the T4 gp59 protein.

  8. Conformation and assembly of polypeptide scaffolds in templating the synthesis of silica: an example of a polylysine macromolecular "switch". (United States)

    Patwardhan, Siddharth V; Maheshwari, Ronak; Mukherjee, Niloy; Kiick, Kristi L; Clarson, Stephen J


    Although the role of polycationic macromolecules in catalyzing the synthesis of silica structures is well established, detailed understanding of the mechanisms behind the production of silica structures of controlled morphologies remains unclear. In this study, we have used both poly-L-lysine (PLL) and/or poly-D-lysine (PDL) for silica synthesis to investigate mechanisms controlling inorganic morphologies. The formation of both spherical silica particles and hexagonal plates was observed. The formation of hexagonal plates was suggested, via circular dichroic spectroscopy (CD), to result from the assembly of helical polylysine molecules. We confirm that the formation of PLL helices is a prerequisite to the hexagonal silica synthesis. In addition, we present for the first time that the handedness of the helicity of the macromolecule does not affect the formation of hexagonal silica. We also show, by using two different silica precursors, that the precursor does not have a direct effect on the formation of hexagonal silica plates. Furthermore, when polylysine helices were converted to beta-sheet structure, only silica particles were obtained, thus suggesting that the adoption of a helical conformation by PLL is required for the formation of hexagonally organized silica. These results demonstrate that the change in polylysine conformation can act as a "switch" in silica structure formation and suggest the potential for controlling morphologies and structures of inorganic materials via control of the conformation of soft macromolecular templates.

  9. Facile synthesis and properties of hierarchical double-walled copper silicate hollow nanofibers assembled by nanotubes. (United States)

    Jin, Renxi; Yang, Yang; Xing, Yan; Chen, Li; Song, Shuyan; Jin, Rongchao


    The hierarchical assembly of multilevel, nonspherical hollow structures remains a considerable challenge. Here, we report a facile approach for synthesizing copper silicate hollow nanofibers with an ultrasmall nanotube-assembled, double-walled structure. The as-prepared hollow fibers possess a tailored complex wall structure, high length-to-diameter ratio, good structural stability, and a high surface area, and they exhibit excellent performance as an easily recycled adsorbent for wastewater treatment and as an ideal support for noble metal catalysts. In addition, this strategy can be extended as a general approach to prepare other double-walled, hollow, fibrous silica-templated materials.

  10. Self-assembly strategies for the synthesis of functional nanostructured materials (United States)

    Perego, M.; Seguini, G.


    Self-assembly is the autonomous organization of components into patterns or structures without human intervention. This is the approach followed by nature to generate living cells and represents one of the practical strategies to fabricate ensembles of nanostructures. In static self-assembly the formation of ordered structures could require energy but once formed the structures are stable. The introduction of additional regular features in the environment could be used to template the self-assembly guiding the organization of the components and determining the final structure they form. In this regard self-assembly of block copolymers represents a potent platform for fundamental studies at the nanoscale and for application-driven investigation as a tool to fabricate functional nanostructured materials. Block copolymers can hierarchically assemble into chemically distinct domains with size and periodicity on the order of 10nm or below, offering a potentially inexpensive route to generate large-area nanostructured materials. The final structure characteristics of these materials are dictated by the properties of the elementary block copolymers, like chain length, volume fraction or degree of block incompatibility. Modern synthetic chemistry offers the possibility to design these macromolecules with very specific length scales and geometries, directly embodying in the block copolymers the code that drives their self- assembling process. The understanding of the kinetics and thermodynamics of the block copolymer self-assembly process in the bulk phase as well as in thin films represents a fundamental prerequisite toward the exploitation of these materials. Incorporating block copolymer into device fabrication procedures or directly into devices, as active elements, will lead to the development of a new generation of devices fabricated using the fundamental law of nature to our advantage in order to minimize cost and power consumption in the fabrication process

  11. Mechanisms of heme utilization by Francisella tularensis.

    Directory of Open Access Journals (Sweden)

    Helena Lindgren

    Full Text Available Francisella tularensis is a highly virulent facultative intracellular pathogen causing the severe disease tularemia in mammals. As for other bacteria, iron is essential for its growth but very few mechanisms for iron acquisition have been identified. Here, we analyzed if and how F. tularensis can utilize heme, a major source of iron in vivo. This is by no means obvious since the bacterium lacks components of traditional heme-uptake systems. We show that SCHU S4, the prototypic strain of subspecies tularensis, grew in vitro with heme as the sole iron source. By screening a SCHU S4 transposon insertion library, 16 genes were identified as important to efficiently utilize heme, two of which were required to avoid heme toxicity. None of the identified genes appeared to encode components of a potential heme-uptake apparatus. Analysis of SCHU S4 deletion mutants revealed that each of the components FeoB, the siderophore system, and FupA, contributed to the heme-dependent growth. In the case of the former two systems, iron acquisition was impaired, whereas the absence of FupA did not affect iron uptake but led to abnormally high binding of iron to macromolecules. Overall, the present study demonstrates that heme supports growth of F. tularensis and that the requirements for the utilization are highly complex and to some extent novel.

  12. Synthesis, double-helix formation, and higher-assembly formation of chiral polycyclic aromatic compounds: conceptual development of polyketide aldol synthesis. (United States)

    Yamaguchi, Masahiko; Shigeno, Masanori; Saito, Nozomi; Yamamoto, Koji


    Polycyclic aromatic compounds are an important group of substances in chemistry, and the study of their properties is a subject of interest in the development of drugs and materials. We have been conducting studies to develop chiral polycyclic aromatic compounds, i.e., helicenes and equatorenes. These helical molecules showed notable aggregate-forming properties and the capability for chiral recognition exerted by noncovalent bond interactions, which were not observed in compounds with central chirality. Homo- and hetero-double-helix-forming helicene oligomers were developed, and the latter self-assembled to form gels and vesicles. In this article, we describe such hierarchical studies of polycyclic aromatic compounds, which were started from polyketide aldol synthesis.

  13. Microwave-assisted click polymerization for the synthesis of A beta(16-22) cyclic oligomers and their self-assembly into polymorphous aggregates

    NARCIS (Netherlands)

    Elgersma, R.C.; van Dijk, M.; Dechesne, A.C.; van Nostrum, C.F.; Hennink, W.E.; Rijkers, D.T.S.; Liskamp, R.M.J.


    We report on the design, synthesis, and structural analysis of cyclic oligomers with an amyloidogenic peptide sequence as the repeating unit to obtain novel self-assembling bionanomaterials. The peptide was derived from the Alzheimer A beta(16-22) sequence since its strong tendency to form antiparal


    For the first time, we report density-assisted self-assembly and efficient synthesis of gold (Au) and platinum (Pt) nanospheres, nanowires and nanorods using vitamin B2 (riboflavin) without employing any special capping or dispersing agent at room temperature; this env...

  15. Synthesis, Characterization, and Secondary Structure Determination of a Silk-Inspired, Self-Assembling Peptide: A Laboratory Exercise for Organic and Biochemistry Courses (United States)

    Albin, Tyler J.; Fry, Melany M.; Murphy, Amanda R.


    This laboratory experiment gives upper-division organic or biochemistry undergraduate students a comprehensive look at the synthesis, chemical characterization, self-assembly, and secondary structure determination of small, N-acylated peptides inspired by the protein structure of silkworm silk. All experiments can be completed in one 4 h lab…

  16. Fatty acid synthesis is inhibited by inefficient utilization of unusual fatty acids for glycerolipid assembly. (United States)

    Bates, Philip D; Johnson, Sean R; Cao, Xia; Li, Jia; Nam, Jeong-Won; Jaworski, Jan G; Ohlrogge, John B; Browse, John


    Degradation of unusual fatty acids through β-oxidation within transgenic plants has long been hypothesized as a major factor limiting the production of industrially useful unusual fatty acids in seed oils. Arabidopsis seeds expressing the castor fatty acid hydroxylase accumulate hydroxylated fatty acids up to 17% of total fatty acids in seed triacylglycerols; however, total seed oil is also reduced up to 50%. Investigations into the cause of the reduced oil phenotype through in vivo [(14)C]acetate and [(3)H]2O metabolic labeling of developing seeds surprisingly revealed that the rate of de novo fatty acid synthesis within the transgenic seeds was approximately half that of control seeds. RNAseq analysis indicated no changes in expression of fatty acid synthesis genes in hydroxylase-expressing plants. However, differential [(14)C]acetate and [(14)C]malonate metabolic labeling of hydroxylase-expressing seeds indicated the in vivo acetyl-CoA carboxylase activity was reduced to approximately half that of control seeds. Therefore, the reduction of oil content in the transgenic seeds is consistent with reduced de novo fatty acid synthesis in the plastid rather than fatty acid degradation. Intriguingly, the coexpression of triacylglycerol synthesis isozymes from castor along with the fatty acid hydroxylase alleviated the reduced acetyl-CoA carboxylase activity, restored the rate of fatty acid synthesis, and the accumulation of seed oil was substantially recovered. Together these results suggest a previously unidentified mechanism that detects inefficient utilization of unusual fatty acids within the endoplasmic reticulum and activates an endogenous pathway for posttranslational reduction of fatty acid synthesis within the plastid.

  17. π-Conjugated aromatics based on truxene: synthesis, self-assembly, and applications. (United States)

    Shi, Ke; Wang, Jie-Yu; Pei, Jian


    Recently, many efforts have been devoted to developing novel polycyclic aromatics due to their unique optical and electronic properties and broad applications, such as in organic field-effect transistors, organic photovoltaics, and organic light-emitting diodes. Among various π-conjugated molecules, many truxene derivatives have interesting characteristics such as C3 -symmetry, strong blue emission, and a planar rigid structure. Moreover, compared with many other π-conjugated aromatics, the synthesis and modification of truxene are particularly facile and diverse. In this account, we summarize investigations into truxene derivatives from synthesis and physical properties to applications in organic electronics.

  18. Design and Synthesis of a Highly Stable Six-hydrogen-bonded Self-assembly Yellowish Green Electroluminescent Molecular Duplex

    Institute of Scientific and Technical Information of China (English)


    This paper describes the design, synthesis and characterization of a hydrogen-bonded molecular duplex with 1,8-naphthalimide fluorescent pendants. The two oligoamide molecular strands, with complementary hydrogen bond sequences of DDADAA and AADADD, caa form an ultra stable self-assembly duplex. Its molecular structure was confirmed by 1H NMR and ESI-MS, and its photoluminescence properties were determined. The resulting duplex exhibited a dramatically enhanced photoluminescence (PL) quantum efficiency of 63.7% compared to the corresponding 1,8-naphthalimide segment (32.4%), suggesting that the formation of the duplex with larger molecular weight could successfully inhibit the quenching of the fluorescent pendant.This novel duplex is a prospective candidate for new electroluminescent emitter.

  19. Selective Synthesis of Molecular Borromean Rings: Engineering of Supramolecular Topology via Coordination-Driven Self-Assembly. (United States)

    Kim, Taegeun; Singh, Nem; Oh, Jihun; Kim, Eun-Hee; Jung, Jaehoon; Kim, Hyunuk; Chi, Ki-Whan


    Molecular Borromean rings (BRs) is one of the rare topology among interlocked molecules. Template-free synthesis of BRs via coordination-driven self-assembly of tetracene-based Ru(II) acceptor and ditopic pyridyl donors is reported. NMR and single-crystal XRD analysis observed sequential transformation of a fully characterized monomeric rectangle to molecular BRs and vice versa. Crystal structure of BRs revealed that the particular topology was enforced by the appropriate geometry of the metallacycle and multiple parallel-displaced π-π interactions between the donor and tetracene moiety of the acceptor. Computational studies based on density functional theory also supported the formation of BRs through dispersive intermolecular interactions in solution.

  20. Self-assembled M2L4 coordination cages : Synthesis and potential applications

    NARCIS (Netherlands)

    Schmidt, Andrea; Casini, Angela; Kuehn, Fritz E.


    Metal-mediated self-assemblies of the general formula MxLy (M = metal ion, L = ligand) have emerged as a promising research area of supramolecular chemistry because of their applicability in various fields such as molecular recognition, catalysis and drug delivery. The focus of this review is on dis

  1. Structure-based design and synthesis of N(omega)-nitro-L-arginine-containing peptidomimetics as selective inhibitors of neuronal nitric oxide synthase. Displacement of the heme structural water. (United States)

    Seo, Jiwon; Igarashi, Jotato; Li, Huiying; Martasek, Pavel; Roman, Linda J; Poulos, Thomas L; Silverman, Richard B


    The neuronal isoform of nitric oxide synthase (nNOS), the enzyme responsible for the production of nitric oxide in the central nervous system, represents an attractive target for the treatment of various neurodegenerative disorders. X-ray crystal structures of complexes of nNOS with two nNOS-selective inhibitors, (4S)-N-{4-amino-5-[(2-aminoethylamino]pentyl}-N'-nitroguanidine (1) and 4-N-(Nomega-nitro-l-argininyl)-trans-4-amino-l-proline amide (2), led to the discovery of a conserved structural water molecule that was hydrogen bonded between the two heme propionates and the inhibitors (Figure 2). On the basis of this observation, we hypothesized that by attaching a hydrogen bond donor group to the amide nitrogen of 2 or to the secondary amine nitrogen of 1, the inhibitor molecules could displace the structural water molecule and obtain a direct interaction with the heme cofactor. To test this hypothesis, peptidomimetic analogues 3-5, which have either an N-hydroxyl (3 and 5) or N-amino (4) donor group, were designed and synthesized. X-ray crystal structures of nNOS with inhibitors 3 and 5 bound verified that the N-hydroxyl group had, indeed, displaced the structural water molecule and provided a direct interaction with the heme propionate moiety (Figures 5 and 6). Surprisingly, in vitro activity assay results indicated that the addition of a hydroxyl group (3) only increased the potency slightly against the neuronal isoform over the parent compound (1). Rationalizations for the small increase in potency are consistent with other changes in the crystal structures.

  2. Synthesis, electrochemistry, STM investigation of oligothiophene self-assemblies with superior structural order and electronic properties (United States)

    Kuo, Cheng-Yu; Liu, Yinghao; Yarotski, Dmitry; Li, Hao; Xu, Ping; Yen, Hung-Ju; Tretiak, Sergei; Wang, Hsing-Lin


    Three oligothiophene (terthiophene, tetrathiophene and pentathiophene) derivatives are synthesized and their monolayer self-assemblies on gold (Au) are prepared via Au-S covalent bond. Our UV-Vis experimental characterization of solution reveals the dependence of the optical properties on the conjugation length of the oligothiophenes, which compares well with Time-Dependent Density Functional Theory (TDDFT) simulations of spectra of individual chromophores. Photoluminescent spectra of thin films show pronounced red shifts compared to that of solutions, suggesting strong inter-oligomer interactions. The comparative studies of cyclic voltammograms of tetrathiophene from solution, cast film and self-assembled monolayer (SAM) indicate presence of one, two, and three oxidized species in these samples, respectively, suggesting a very strong electronic coupling between tetrathiophene molecules in the SAM. Scanning tunneling microscopy (STM) imaging of SAMs of the tetrathiophene on an atomically flat Au surface exhibits formation of monolayer assemblies with molecular order, and the molecular packing appears to show an overlay of oligothiophene molecules on top of another one. In contrast, the trimer and pentamer images show only aggregated species lacking long-range order on the molecular level. Such trends in going from disordered-ordered-disordered monolayer assemblies are mainly due to a delicate balance between inter-chromophore π-π couplings, hydrophobic interaction and the propensity to form Au-S covalent bond. Such hypothesis has been validated by our computational results suggesting different interaction patterns of oligothiophenes with odd numbered and even numbered thiophene repeat units placed in a dimer configuration. Observed correlations between oligomer geometry and structural order of monolayer assembly elucidate important structure-property relationships and have implications for these molecular structures in organic optoelectronic devices and energy

  3. Synthesis of GoldMag particles with assembled structure and their applications in immunoassay

    Institute of Scientific and Technical Information of China (English)

    CUI; Yali; ZHANG; Lianying; SU; Jing; ZHANG; Caifeng; LI; Qi; CUI; Ting; JIN; Boquan; CHEN; Chao


    Micrometer-sized Fe3O4 particles and nano-sized gold particles were first synthesized by methods of self-aggregation of surface-chemically modified Fe3O4 nanoparticles and citrate reduction of the Au3+ to Au0, respectively. Interaction between these two types of particles resulted in the assembly of nano-sized gold particles on the surface of the micrometer-sized Fe3O4 particles, forming an assembled structure with the Fe3O4 core particles around which are attached nano-sized gold particles. The Fe3O4/Au structure is named GoldMag particles with assembled structure. The synthetic process, structure, and magnetic property of the GoldMag particles were analyzed. GoldMag particles with assembled structure have an irregular shape, rough surface with a diameter of 2-3 (m. These particles exhibit the superparamagnetic property with saturated magnetization of 41 A·m2/kg. In a single step, antibodies could be readily immobilized onto the surface of the particles with a high binding capacity. The GoldMag particles can be used as a novel carrier in immunoassays. The maximum quantity of human IgG immobilized onto GoldMag particles was 330 (g/mg. In order to validate the quality of the GoldMag particles as immunoassay carriers, an immunoassay system was used. The relative amount of immobilized human IgG was measured by HRP-labeled anti human IgG. The coefficient of variation within parallel samples of each group was below 6% and the coefficient of variation of means between five groups carried out separately was below 7%. Based on the sandwich method, the Hepatitis B surface antigen (HBsAg) and interleukin-8 (IL-8) were also analyzed by qualitative and quantitative detection, respectively. The result indicated that the GoldMag particles with assembled structure were an ideal carrier in immunoassay.

  4. Salicylic acid induces mitochondrial injury by inhibiting ferrochelatase heme biosynthesis activity. (United States)

    Gupta, Vipul; Liu, Shujie; Ando, Hideki; Ishii, Ryohei; Tateno, Shumpei; Kaneko, Yuki; Yugami, Masato; Sakamoto, Satoshi; Yamaguchi, Yuki; Nureki, Osamu; Handa, Hiroshi


    Salicylic acid is a classic nonsteroidal anti-inflammatory drug. Although salicylic acid also induces mitochondrial injury, the mechanism of its antimitochondrial activity is not well understood. In this study, by using a one-step affinity purification scheme with salicylic acid-immobilized beads, ferrochelatase (FECH), a homodimeric enzyme involved in heme biosynthesis in mitochondria, was identified as a new molecular target of salicylic acid. Moreover, the cocrystal structure of the FECH-salicylic acid complex was determined. Structural and biochemical studies showed that salicylic acid binds to the dimer interface of FECH in two possible orientations and inhibits its enzymatic activity. Mutational analysis confirmed that Trp301 and Leu311, hydrophobic amino acid residues located at the dimer interface, are directly involved in salicylic acid binding. On a gel filtration column, salicylic acid caused a shift in the elution profile of FECH, indicating that its conformational change is induced by salicylic acid binding. In cultured human cells, salicylic acid treatment or FECH knockdown inhibited heme synthesis, whereas salicylic acid did not exert its inhibitory effect in FECH knockdown cells. Concordantly, salicylic acid treatment or FECH knockdown inhibited heme synthesis in zebrafish embryos. Strikingly, the salicylic acid-induced effect in zebrafish was partially rescued by FECH overexpression. Taken together, these findings illustrate that FECH is responsible for salicylic acid-induced inhibition of heme synthesis, which may contribute to its antimitochondrial and anti-inflammatory function. This study establishes a novel aspect of the complex pharmacological effects of salicylic acid.

  5. Expeditious organic–free assembly: morphologically controlled synthesis of iron oxides using microwaves (United States)

    A microwave hydrothermal method is developed for the synthesis of iron oxides, α-Fe2O3, β-FeOOH, and the junction of α-Fe2O3–β-FeOOH. This method is absolutely organic-free, and various structures could be obtained simply by changing th...

  6. Assembling Synthesis of BaSO4 Biomimetic Nano-superstructures through Eggshell Membrane Template

    Institute of Scientific and Technical Information of China (English)

    LIU Jin-ku; WU Qing-sheng; DING Ya-ping


    @@ Introduction The controlled synthesis of inorganic materials with specific sizes,architectures and morphologies has attracted intensive interest due to their importance in fundamental research and the potential brought out by them to design new materials and devices in various fields such as medicine,electronics, ceramics and pigments[1-4].

  7. Differential effects of heme oxygenase isoforms on heme mediation of endothelial intracellular adhesion molecule 1 expression. (United States)

    Wagener, F A; da Silva, J L; Farley, T; de Witte, T; Kappas, A; Abraham, N G


    Heme oxygenase (HO), by catabolizing heme to bile pigments, down-regulates cellular hemoprotein, hemoglobin, and heme; the latter generates pro-oxidant products, including free radicals. Two HO isozymes, the products of distinct genes, have been described; HO-1 is the inducible isoform, whereas HO-2 is suggested to be constitutively expressed. We studied the inducing effect of several metal compounds (CoCl(2), stannic mesoporphyrin, and heme) on HO activity. Additionally, we studied HO-1 expression in experimental models of adhesion molecule expression produced by heme in endothelial cells, and the relationship of HO-1 expression to the induced adhesion molecules. Flow cytometry analysis showed that heme induces intracellular adhesion molecule 1 (ICAM-1) expression in a concentration (10-100 microM)- and time (1-24 h)-dependent fashion in human umbilical vein endothelial cells. Pretreatment with stannic mesoporphyrin, an inhibitor of HO activity, caused a 2-fold increase in heme-induced ICAM-1 expression. In contrast, HO induction by CoCl(2) decreased heme-induced ICAM-1 expression by 33%. To examine the contribution of HO-1 and HO-2 to endothelial HO activity, specific antisense oligonucleotides (ODNs) of each isoform were tested for their specificity to inhibit HO activity in cells exposed to heme. Endothelial cells exposed to heme elicited increased HO activity, which was prevented (70%) by HO-1 antisense ODNs. HO-2 antisense ODN inhibited heme-induced HO activity by 21%. Addition of HO-1 antisense ODNs prevented heme degradation and resulted in elevation of microsomal heme. Western blot analysis showed that HO-1 antisense ODNs selectively inhibited HO-1 protein and failed to inhibit HO-2 protein. Incubation of endothelial cells with HO-1 antisense enhanced heme-dependent increase of ICAM-1. In contrast, addition of HO-2 antisense to endothelial cells failed to increase adhesion molecules. The role of glutathione, an important antioxidant, was examined on heme

  8. Synthesis and characterization of novel antibacterial silver nanocomposite nanofiltration and forward osmosis membranes based on layer-by-layer assembly. (United States)

    Liu, Xin; Qi, Saren; Li, Ye; Yang, Liang; Cao, Bin; Tang, Chuyang Y


    Using layer-by-layer (LbL) assembly method, we fabricated novel silver nanocomposite LbL-Ag nanofiltration (NF) and forward osmosis (FO) membranes. The incorporation of silver nanoparticles (AgNPs) in the membranes did not adversely affect the membrane separation performance in NF and FO processes at low AgNPs incorporation levels (0.22-1.19 wt.% as silver). The FO performance of the xLbL-Ag membranes was better than or comparable to most NF-like FO membranes reported in the literature. In addition, the silver nanocomposite membranes exhibited excellent antibacterial properties against both Gram-positive Bacillus subtilis and Gram-negative Escherichia coli. Our results showed that the performances of the silver nanocomposite membranes are highly dependent on silver incorporation in the membranes, which could be controlled by using different membrane synthesis routines and doping of AgNPs. To the best knowledge of the authors, this is the first study on fabrication and characterization of novel antibacterial silver nanocomposite NF and FO membranes through LbL assembly approach.

  9. Heme sensing in Bacillus thuringiensis: a supplementary HssRS-regulated heme resistance system. (United States)

    Schmidt, Rachel M; Carter, Micaela M; Chu, Michelle L; Latario, Casey J; Stadler, Sarah K; Stauff, Devin L


    Several Gram-positive pathogens scavenge host-derived heme to satisfy their nutritional iron requirement. However, heme is a toxic molecule capable of damaging the bacterial cell. Gram-positive pathogens within the phylum Firmicutes overcome heme toxicity by sensing heme through HssRS, a two-component system that regulates the heme detoxification transporter HrtAB. Here we show that heme sensing by HssRS and heme detoxification by HrtAB occur in the insect pathogen Bacillus thuringiensis We find that in B. thuringiensis, HssRS directly regulates an operon, hrmXY, encoding hypothetical membrane proteins that are not found in other Firmicutes with characterized HssRS and HrtAB systems. This novel HssRS-regulated operon or its orthologs BMB171_c3178 and BMB171_c3330 are required for maximal heme resistance. Furthermore, the activity of HrmXY is not dependent on expression of HrtAB. These results suggest that B. thuringiensis senses heme through HssRS and induces expression of separate membrane-localized systems capable of overcoming different aspects of heme toxicity.

  10. Self-assembly versus stepwise synthesis: heterometal-organic frameworks based on metalloligands with tunable luminescence properties. (United States)

    Zhang, Shu-Ran; Du, Dong-Ying; Tan, Ke; Qin, Jun-Sheng; Dong, Hui-Qing; Li, Shun-Li; He, Wen-Wen; Lan, Ya-Qian; Shen, Ping; Su, Zhong-Min


    A new family of heterometal-organic frameworks has been prepared by two synthesis strategies, in which IFMC-26 and IFMC-27 are constructed by self-assembly and IFMC-28 is obtained by stepwise synthesis based on the metalloligand (IFMC=Institute of Functional Material Chemistry). IFMC-26 is a (3,6)-connected net and IFMC-27 is a (4,8)-connected 3D framework. The metalloligands {Ni(H4 L)}(NO3 )2 are connected by binuclear lanthanide clusters giving rise to a 2D sheet structure in IFMC-28. Notably, IFMC-26-Eux Tby and IFMC-28-Eux Tby have been obtained by changing the molar ratios of raw materials. Owing to the porosity of IFMC-26, Tb(3+) @IFMC-26-Eu and Eu(3+) @IFMC-26-Tb are obtained by postencapsulating Tb(III) and Eu(III) ions into the pores, respectively. Tunable luminescence in metal-organic frameworks is achieved by the two kinds of doping methods. In particular, the quantum yields of heterometal-organic frameworks are apparently enhanced by postencapsulation of Ln(III) ions.

  11. Block copolymer self-assembly-directed synthesis of mesoporous gyroidal superconductors. (United States)

    Robbins, Spencer W; Beaucage, Peter A; Sai, Hiroaki; Tan, Kwan Wee; Werner, Jörg G; Sethna, James P; DiSalvo, Francis J; Gruner, Sol M; Van Dover, Robert B; Wiesner, Ulrich


    Superconductors with periodically ordered mesoporous structures are expected to have properties very different from those of their bulk counterparts. Systematic studies of such phenomena to date are sparse, however, because of a lack of versatile synthetic approaches to such materials. We demonstrate the formation of three-dimensionally continuous gyroidal mesoporous niobium nitride (NbN) superconductors from chiral ABC triblock terpolymer self-assembly-directed sol-gel-derived niobium oxide with subsequent thermal processing in air and ammonia gas. Superconducting materials exhibit a critical temperature (T c) of about 7 to 8 K, a flux exclusion of about 5% compared to a dense NbN solid, and an estimated critical current density (J c) of 440 A cm(-2) at 100 Oe and 2.5 K. We expect block copolymer self-assembly-directed mesoporous superconductors to provide interesting subjects for mesostructure-superconductivity correlation studies.

  12. Confined cooperative self-assembly and synthesis of optically and electrically active nanostructures : final LDRD report

    Energy Technology Data Exchange (ETDEWEB)

    Coker, Eric Nicholas; Haddad, Raid Edward (University of New Mexico, Albuquerque, NM); Fan, Hongyou; Ta, Anh (University of New Mexico, Albuquerque, NM); Bai, Feng (University of New Mexico, Albuquerque, NM); Rodriguez, Mark Andrew; Huang, Jian Yu


    In this project, we developed a confined cooperative self-assembly process to synthesize one-dimensional (1D) j-aggregates including nanowires and nanorods with controlled diameters and aspect ratios. The facile and versatile aqueous solution process assimilates photo-active macrocyclic building blocks inside surfactant micelles, forming stable single-crystalline high surface area nanoporous frameworks with well-defined external morphology defined by the building block packing. Characterizations using TEM, SEM, XRD, N{sub 2} and NO sorption isotherms, TGA, UV-vis spectroscopy, and fluorescence imaging and spectroscopy indicate that the j-aggregate nanostructures are monodisperse and may further assemble into hierarchical arrays with multi-modal functional pores. The nanostructures exhibit enhanced and collective optical properties over the individual chromophores. This project was a small footprint research effort which, nonetheless, produced significant progress towards both the stated goal as well as unanticipated research directions.

  13. Synthesis and self-assembly of photonic materials from nanocrystalline titania sheets. (United States)

    Zhu, Jian; Wang, Jinguo; Lv, Fujian; Xiao, Shengxiong; Nuckolls, Colin; Li, Hexing


    We describe the use of benzyl alcohols in a solvothermal/alcoholysis reaction to form nanocrystalline sheets of anatase titania. By tuning the reaction conditions, we adjust the size of the nanosheets. The type and density of benzyl groups that decorate the basal plane of the titania sheets control the self-assembly into layered structures. These layered materials can be grown from solid substrates to create iridescent thin films that reflect specific wavelengths of visible light.

  14. Self-assembled polyhydroxy fatty acids vesicles: a mechanism for plant cutin synthesis. (United States)

    Heredia-Guerrero, José A; Benítez, José J; Heredia, Antonio


    Despite its biological importance, the mechanism of formation of cutin, the polymeric matrix of plant cuticles, has not yet been fully clarified. Here, for the first time, we show the participation in the process of lipid vesicles formed by the self-assembly of endogenous polyhydroxy fatty acids. The accumulation and fusion of these vesicles (cutinsomes) at the outer part of epidermal cell wall is proposed as the mechanism for early cuticle formation.

  15. Development of Self-Assembled Supramolecular Catalysts and Their Applications to Organic Synthesis

    Institute of Scientific and Technical Information of China (English)

    Shiro IKEGAMI


    @@ 1Introduction In modern synthetic organic chemistry, the development of efficient reagent or catalyst recycling systems is regarded as one of the most important topics. We have previously reported a self-assembled process between poly(N-isopropylacrylamide) (PNIPAAm) based polymer ligands and an inorganic species[1]. This process afforded a networked supramolecular complex where the polymers are cross-linked together by the inorganic species. Thus obtained complex was insoluble in water and worked as an efficient triphase catalyst.

  16. Synthesis, optical properties, and helical self-assembly of a bivaline-containing tetraphenylethene (United States)

    Li, Hongkun; Zheng, Xiaoyan; Su, Huimin; Lam, Jacky W. Y.; Sing Wong, Kam; Xue, Shan; Huang, Xuejiao; Huang, Xuhui; Li, Bing Shi; Tang, Ben Zhong


    A chiral tetraphenylethene derivative with two valine-containing attachments (TPE-DVAL), was synthesized by Cu(I)-catalyzed azide-alkyne “click” reaction. The optical properties and self-assembling behaviours of TPE-DVAL were investigated. The molecule is non-emissive and circular dichroism (CD)-silent in solution, but shows strong fluorescence and Cotton effects in the aggregation state, demonstrating aggregation-induced emission (AIE) and CD (AICD) characteristics. TPE-DVAL exhibits good circularly polarized luminescence (CPL) when depositing on the surface of quartz to allow the evaporation of its 1,2-dichloroethane solution. SEM and TEM images of the molecule show that the molecule readily self-assembles into right-handed helical nanofibers upon the evaporation of its solvent of DCE. The molecular alignments and interactions in assembling process are further explored through XRD analysis and computational simulation. The driving forces for the formation of the helical fibers were from the cooperative effects of intermolecular hydrogen bonding, π-π interactions and steric effect.

  17. Supramolecular assembled three-dimensional graphene hybrids: Synthesis and applications in supercapacitors (United States)

    Ni, Lubin; Zhang, Wang; Wu, Zhen; Sun, Chunyu; Cai, Yin; Yang, Guang; Chen, Ming; Piao, Yuanzhe; Diao, Guowang


    Graphene-based materials have received worldwide attention in the focus of forefront energy storage investigations. Currently, the design of novel three-dimensional (3D) graphene structures with high energy capability, superior electron and ion conductivity, and robust mechanical flexibility is still a great challenge. Herein, we have successfully demonstrated a novel approach to fabricate 3D assembled graphene through the supramolecular interactions of β-cyclodextrin polymers (β-CDP) with an adamantine end-capped poly(ethylene oxide) polymer linker (PEG-AD). The incorporation of PEG-AD linker into rGO sheets increased the interlayer spacing of rGO sheets to form 3D graphene materials, which can provide efficient 3D electron transfer pathways and ion diffusion channels, and facilitate the infiltration of gel electrolyte. The as-prepared 3D self-assembled graphene materials exhibit significantly improved electrochemical performances of supercapacitor in terms of high specific capacitance, remarkable rate capability, and excellent cycling stability compared to pristine reduced graphene oxide. This study shed new lights to the construction of three dimensional self-assembled graphene materials and their urgent applications in energy storage.

  18. Precise positioning and compliance synthesis for automatic assembly using Lorentz levitation (United States)

    Hollis, R. L.; Salcudean, S.


    Many manufacturing assembly tasks require fine compliant motion and fast, accurate positioning. Conventional robots perform poorly in these tasks because of their large mass, friction and backlash in gears, cogging in drive motors and other deleterious effects. Even robots equipped with special control systems enabling compliant operation offer only partial solutions. It is difficult or impossible to automate many product assemblies requiring fine, compliant motion. This problem can be greatly alleviated by dividing the manipulation system into coarse and fine domains. In this scenario, a standard industrial robot can serve as a coarse positioner which in turn carries a six degrees of freedom fine motion wrist. Thus the robot can access a workspace measured in meters at low bandwidth and low resolution while the wrist can move over millimeters at high bandwidth and high resolution during the final phase of the assembly operation. Work indicates that fine motion wrists using Lorentz levitation can greatly augment the accuracy and dexterity of robots because they are frictionless, have high bandwidths and have a single back drivable moving part. Also, since there is no contact between the moving and stationary parts, wear and contamination can be eliminated. The use of six Lorentz force actuators in combination with real time position and orientation sensing offers several important advantages over magnetic bearing approaches.

  19. Influence of heme-thiolate in shaping the catalytic properties of a bacterial nitric-oxide synthase. (United States)

    Hannibal, Luciana; Somasundaram, Ramasamy; Tejero, Jesús; Wilson, Adjele; Stuehr, Dennis J


    Nitric-oxide synthases (NOS) are heme-thiolate enzymes that generate nitric oxide (NO) from L-arginine. Mammalian and bacterial NOSs contain a conserved tryptophan (Trp) that hydrogen bonds with the heme-thiolate ligand. We mutated Trp(66) to His and Phe (W66H, W66F) in B. subtilis NOS to investigate how heme-thiolate electronic properties control enzyme catalysis. The mutations had opposite effects on heme midpoint potential (-302, -361, and -427 mV for W66H, wild-type (WT), and W66F, respectively). These changes were associated with rank order (W66H < WT < W66F) changes in the rates of oxygen activation and product formation in Arg hydroxylation and N-hydroxyarginine (NOHA) oxidation single turnover reactions, and in the O(2) reactivity of the ferrous heme-NO product complex. However, enzyme ferrous heme-O(2) autoxidation showed an opposite rank order. Tetrahydrofolate supported NO synthesis by WT and the mutant NOS. All three proteins showed similar extents of product formation (L-Arg → NOHA or NOHA → citrulline) in single turnover studies, but the W66F mutant showed a 2.5 times lower activity when the reactions were supported by flavoproteins and NADPH. We conclude that Trp(66) controls several catalytic parameters by tuning the electron density of the heme-thiolate bond. A greater electron density (as in W66F) improves oxygen activation and reactivity toward substrate, but decreases heme-dioxy stability and lowers the driving force for heme reduction. In the WT enzyme the Trp(66) residue balances these opposing effects for optimal catalysis.

  20. Heme environment in HmuY, the heme-binding protein of Porphyromonas gingivalis

    Energy Technology Data Exchange (ETDEWEB)

    Wojtowicz, Halina [Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw, Tamka 2, 50-137 Wroclaw (Poland); Wojaczynski, Jacek [Department of Chemistry, University of Wroclaw, 50-383 Wroclaw (Poland); Olczak, Mariusz [Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw, Tamka 2, 50-137 Wroclaw (Poland); Kroliczewski, Jaroslaw [Laboratory of Biophysics, Faculty of Biotechnology, University of Wroclaw, 50-148 Wroclaw (Poland); Latos-Grazynski, Lechoslaw [Department of Chemistry, University of Wroclaw, 50-383 Wroclaw (Poland); Olczak, Teresa, E-mail: [Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw, Tamka 2, 50-137 Wroclaw (Poland)


    Porphyromonas gingivalis, a Gram-negative anaerobic bacterium implicated in the development and progression of chronic periodontitis, acquires heme for growth by a novel mechanism composed of HmuY and HmuR proteins. The aim of this study was to characterize the nature of heme binding to HmuY. The protein was expressed, purified and detailed investigations using UV-vis absorption, CD, MCD, and {sup 1}H NMR spectroscopy were carried out. Ferric heme bound to HmuY may be reduced by sodium dithionite and re-oxidized by potassium ferricyanide. Heme complexed to HmuY, with a midpoint potential of 136 mV, is in a low-spin Fe(III) hexa-coordinate environment. Analysis of heme binding to several single and double HmuY mutants with the methionine, histidine, cysteine, or tyrosine residues replaced by an alanine residue identified histidines 134 and 166 as potential heme ligands.

  1. Transcriptional profile of Haemophilus influenzae: effects of iron and heme. (United States)

    Whitby, Paul W; Vanwagoner, Timothy M; Seale, Thomas W; Morton, Daniel J; Stull, Terrence L


    Haemophilus influenzae requires either heme or a porphyrin and iron source for growth. Microarray studies of H. influenzae strain Rd KW20 identified 162 iron/heme-regulated genes, representing approximately 10% of the genome, with > or =1.5-fold changes in transcription in response to iron/heme availability in vitro. Eighty genes were preferentially expressed under iron/heme restriction; 82 genes were preferentially expressed under iron/heme-replete conditions.

  2. Utility of heme analogues to intentionally modify heme-globin interactions in myoglobin. (United States)

    Neya, Saburo; Nagai, Masako; Nagatomo, Shigenori; Hoshino, Tyuji; Yoneda, Tomoki; Kawaguchi, Akira T


    Myoglobin reconstitution with various synthetic heme analogues was reviewed to follow the consequences of modified heme-globin interactions. Utility of dimethyl sulfoxide as the solvent for water-insoluble hemes was emphasized. Proton NMR spectroscopy revealed that loose heme-globin contacts in the heme pocket eventually caused the dynamic heme rotation around the iron-histidine bond. The full rotational rate was estimated to be about 1400 s(-1) at room temperature for 1,4,5,8-tetramethylhemin. The X-ray analysis of the myoglobin containing iron porphine, the smallest heme without any side chains, showed that the original globin fold was well conserved despite the serious disruption of native heme-globin contacts. Comparison between the two myoglobins with static and rotatory prosthetic groups indicated that the oxygen and carbon monoxide binding profiles were almost unaffected by the heme motion. On the other hand, altered tetrapyrrole array of porphyrin dramatically changed the dissociation constant of oxygen from 0.0005 mm Hg of porphycene-myoglobin to ∞ in oxypyriporphyrin-myoglobin. Heme-globin interactions in myoglobin were also monitored with circular dichroism spectroscopy. The observation on several reconstituted protein revealed an unrecognized role of the propionate groups in protoheme. Shortening of heme 6,7-propionates to carboxylates resulted in almost complete disappearance of the positive circular dichroism band in the Soret region. The theoretical analysis suggested that the disappeared circular dichroism band reflected the cancellation effects between different conformers of the carboxyl groups directly attached to heme periphery. The above techniques were proposed to be applicable to other hemoproteins to create new biocatalysts. This article is part of a Special Issue entitled Biodesign for Bioenergetics--the design and engineering of electronic transfer cofactors, proteins and protein networks, edited by Ronald L. Koder and J.L. Ross Anderson.

  3. Synthesis and self-assembly of thio derivatives of calix[4]arene on noble metal surfaces. (United States)

    Genorio, Bostjan; He, Tao; Meden, Anton; Polanc, Slovenko; Jamnik, Janko; Tour, James M


    Self-assembled monolayers (SAMs) provide a simple route to functionalize electrode surfaces with organic molecules. Herein we use cavity-containing derivatives of calix[4]arenes in SAMs. Bound to noble metal surface, the assembled molecules are candidates to serve as molecular sieves for H 2 molecules and H (+) ions, which could have relevance for fuel cell applications. Tetra- O-alkylated calix[4]arenes with thiolacetate and thiolamide wide-rim anchoring groups in cone and partial-cone conformations were designed, synthesized and self-assembled onto Au, Pt, and Pd surfaces. The resulting SAMs were systematically examined. Single crystal X-ray diffraction of 5,11,17,23-tetrakis(thioacetyl)-25,26,27,28-tetra- i-propoxycalix[4]arene confirmed the cone conformation and revealed the cavity dimensions of the SAMs that were formed by immersing noble metal substrates (Au, Pt and Pd deposited on Si-wafers) in solutions of calix[4]arenes. Surface characterization techniques including ellipsometry, cyclic voltammetry (CV) and X-ray photoelectron spectroscopy (XPS) were used, indicating that the metal surface is terminated with a monomolecular layer. Experimental thicknesses obtained from the ellipsometry are consistent with the calculated values. CV results showed 50 to 80% physical passivation against the Fe(CN) 6 (3-/4-) couple, implying an overall relatively low concentration of defects and pinholes in the films. The binding energies of the S2p core level in the XPS were consistent with the literature values and revealed that up to 3.2 out of four anchoring groups were bonded to the noble metal surface.

  4. Synthesis of Self-Assembled Multifunctional Nanocomposite Catalysts with Highly Stabilized Reactivity and Magnetic Recyclability (United States)

    Yu, Xu; Cheng, Gong; Zheng, Si-Yang


    In this paper, a multifunctional Fe3O4@SiO2@PEI-Au/Ag@PDA nanocomposite catalyst with highly stabilized reactivity and magnetic recyclability was synthesized by a self-assembled method. The magnetic Fe3O4 nanoparticles were coated with a thin layer of the SiO2 to obtain a negatively charged surface. Then positively charged poly(ethyleneimine) polymer (PEI) was self-assembled onto the Fe3O4@SiO2 by electrostatic interaction. Next, negatively charged glutathione capped gold nanoparticles (GSH-AuNPs) were electrostatically self-assembled onto the Fe3O4@SiO2@PEI. After that, silver was grown on the surface of the nanocomposite due to the reduction of the dopamine in the alkaline solution. An about 5 nm thick layer of polydopamine (PDA) was observed to form the Fe3O4@SiO2@PEI-Au/Ag@PDA nanocomposite. The Fe3O4@SiO2@PEI-Au/Ag@PDA nanocomposite was carefully characterized by the SEM, TEM, FT-IR, XRD and so on. The Fe3O4@SiO2@PEI-Au/Ag@PDA nanocomposite shows a high saturation magnetization (Ms) of 48.9 emu/g, which allows it to be attracted rapidly to a magnet. The Fe3O4@SiO2@PEI-Au/Ag@PDA nanocomposite was used to catalyze the reduction of p-nitrophenol (4-NP) to p-aminophenol (4-AP) as a model system. The reaction kinetic constant k was measured to be about 0.56 min‑1 (R2 = 0.974). Furthermore, the as-prepared catalyst can be easily recovered and reused for 8 times, which didn’t show much decrease of the catalytic capability.

  5. In-Cell Enzymology To Probe His-Heme Ligation in Heme Oxygenase Catalysis. (United States)

    Sigala, Paul A; Morante, Koldo; Tsumoto, Kouhei; Caaveiro, Jose M M; Goldberg, Daniel E


    Heme oxygenase (HO) is a ubiquitous enzyme with key roles in inflammation, cell signaling, heme disposal, and iron acquisition. HO catalyzes the oxidative conversion of heme to biliverdin (BV) using a conserved histidine to coordinate the iron atom of bound heme. This His-heme interaction has been regarded as being essential for enzyme activity, because His-to-Ala mutants fail to convert heme to biliverdin in vitro. We probed a panel of proximal His mutants of cyanobacterial, human, and plant HO enzymes using a live-cell activity assay based on heterologous co-expression in Escherichia coli of each HO mutant and a fluorescent biliverdin biosensor. In contrast to in vitro studies with purified proteins, we observed that multiple HO mutants retained significant activity within the intracellular environment of bacteria. X-ray crystallographic structures of human HO1 H25R with bound heme and additional functional studies suggest that HO mutant activity inside these cells does not involve heme ligation by a proximal amino acid. Our study reveals unexpected plasticity in the active site binding interactions with heme that can support HO activity within cells, suggests important contributions by the surrounding active site environment to HO catalysis, and can guide efforts to understand the evolution and divergence of HO function.

  6. Spectroscopic characterization of a higher plant heme oxygenase isoform-1 from Glycine max (soybean)--coordination structure of the heme complex and catabolism of heme. (United States)

    Gohya, Tomohiko; Zhang, Xuhong; Yoshida, Tadashi; Migita, Catharina T


    Heme oxygenase converts heme into biliverdin, CO, and free iron. In plants, as well as in cyanobacteria, heme oxygenase plays a particular role in the biosynthesis of photoreceptive pigments, such as phytochromobilins and phycobilins, supplying biliverdin IX(alpha) as a direct synthetic resource. In this study, a higher plant heme oxygenase, GmHO-1, of Glycine max (soybean), was prepared to evaluate the molecular features of its heme complex, the enzymatic activity, and the mechanism of heme conversion. The similarity in the amino acid sequence between GmHO-1 and heme oxygenases from other biological species is low, and GmHO-1 binds heme with 1 : 1 stoichiometry at His30; this position does not correspond to the proximal histidine of other heme oxygenases in their sequence alignments. The heme bound to GmHO-1, in the ferric high-spin state, exhibits an acid-base transition and is converted to biliverdin IX(alpha) in the presence of NADPH/ferredoxin reductase/ferredoxin, or ascorbate. During the heme conversion, an intermediate with an absorption maximum different from that of typical verdoheme-heme oxygenase or CO-verdoheme-heme oxygenase complexes was observed and was extracted as a bis-imidazole complex; it was identified as verdoheme. A myoglobin mutant, H64L, with high CO affinity trapped CO produced during the heme degradation. Thus, the mechanism of heme degradation by GmHO-1 appears to be similar to that of known heme oxygenases, despite the low sequence homology. The heme conversion by GmHO-1 is as fast as that by SynHO-1 in the presence of NADPH/ferredoxin reductase/ferredoxin, thereby suggesting that the latter is the physiologic electron-donating system.

  7. Synthesis and Self-Assembly of Donor–Acceptor–Donor Based Oligothiophenes and Their Optoelectronic Properties

    DEFF Research Database (Denmark)

    Siram, Raja Bhaskar Kanth; Tandy, Kristen; Horecha, Marta;


    In this work, the synthesis of an oligothiophene having a donor–acceptor–donor (D–A–D) chromophore with hydrogen bonding groups is described. The D–A–D molecule was demonstrated to self-organize via intermolecular H-bonding between barbituric acid units. Interactions between the oligothiophene...... subunits were also found to be important, affording nanoribbons that could be observed by atomic force and transmission electron microscopy. The applicability of the oligothiophene for organic electronic applications was investigated by fabricating organic field-effect transistors (OFETs) and organic...

  8. Synthesis of D-Desosamine and Analogs by Rapid Assembly of 3-Amino Sugars. (United States)

    Zhang, Ziyang; Fukuzaki, Takehiro; Myers, Andrew G


    D-Desosamine is synthesized in 4 steps from methyl vinyl ketone and sodium nitrite. The key step in this chromatography-free synthesis is the coupling of (R)-4-nitro-2-butanol and glyoxal (trimeric form) mediated by cesium carbonate, which affords in crystalline form 3-nitro-3,4,6-trideoxy-α-D-glucose, a nitro sugar stereochemically homologous to D-desosamine. This strategy has enabled the syntheses of an array of analogous 3-nitro sugars. In each case the 3-nitro sugars are obtained in pure form by crystallization.


    Institute of Scientific and Technical Information of China (English)

    Jing Wang; Yuan Yao; Bing Ji; Wei Huang; Yong-feng Zhou; De-yue Yan


    The amphiphilic multiarm copolymers were synthesized through the modification of commercially available hyperbranched polyesters (Boltom H40) with N-ε-carbobenzoxy-L-Lysine N-carboxyanhydride (ZLys-NCA). After being condensed with N-Boc-phenylalanine (Boc-NPhe) and deprotected the Boc-groups in trifluoroacetic acid (TFA), the original terminal hydroxyl groups were transformed into the amino groups and then initiated the ring-opening polymerization of ZLys-NCA. The hydrophilic poly(L-lysine) was grafted to the surface of Boltorn H40 successfully after the protecting benzyl groups were removed by the HBr solution in glacial acetic acid (33 wt%). The resulting multiarm copolymers were characterized by the 1H-NMR, GPC and FTIR. The arm length calculated by NMR and GPC analysis was about 3 and 13 lysine-units for H40-Phe-PLysl and H40-Phe-PLys2 respectively. Due to the amphiphilic molecular structure, they displayed ability to self-assemble into spherical micelles in aqueous solution with the average diameter in the range from 70 nm to 250 nm. The CMC of H40-Phe-PLysl and H40-Phe-PLys2 was 0.013 mg/mL and 0.028 mg/mL, respectively,indicating that H40-Phe-PLysl with shorter arm length is easier to self-assemble than H40-Phe-PLys2 with longer arm length.

  10. Synthesis and Self-assembling of Coumarin-based Low-molecular Weight Organogelators

    Institute of Scientific and Technical Information of China (English)

    CHEN Guo-jun; XUE Peng-chong; LU Ran; SONG Dong-po; BAO Chun-yan; XU Ting-hua; ZHAO Ying-ying


    Four coumarin derivatives(4a-4d) with different alkoxy chains were synthesized.It was found that compound 4d showed a better gelation ability than the other compounds,for example,it could self-assemble into organogeis in various organic fluids via ultrasound treatment or heating-cooling process,whereas compound 4c could only gel in a few mixed solvents and compounds 4a,4b could not form organogel.The results from fluorescent and F%IR spectra indicate that π-π interaction had an effect on the formation of the organogels of compound 4d besides H-bonding and van der Waals interaction,which were the driving forces for the self-assembling of compound 4c in gel state.The gel of compound 4d in toluene could emit strong fluorescence under UV irradiation and the [2+2]cyclo-addition was suggested by 1H NMR and fluorescence spectroscopy.This light-sensitive organogel might find application in optical materials.

  11. Synthesis and Characterization of Self-Assembled Nanogels Made of Pullulan

    Directory of Open Access Journals (Sweden)

    Sílvia A. Ferreira


    Full Text Available Self-assembled nanogels made of hydrophobized pullulan were obtained using a versatile, simple, reproducible and low-cost method. In a first reaction pullulan was modified with hydroxyethyl methacrylate or vinyl methacrylate, further modified in the second step with hydrophobic 1-hexadecanethiol, resulting as an amphiphilic material, which self-assembles in water via the hydrophobic interaction among alkyl chains. Structural features, size, shape, surface charge and stability of the nanogels were studied using hydrogen nuclear magnetic resonance, fluorescence spectroscopy, cryo-field emission scanning electron microscopy and dynamic light scattering. Above the critical aggregation concentration spherical polydisperse macromolecular micelles revealed long-term colloidal stability in aqueous medium, with a nearly neutral negative surface charge and mean hydrodynamic diameter in the range 100–400 nm, depending on the polymer degree of substitution. Good size stability was observed when nanogels were exposed to potential destabilizing pH conditions. While the size stability of the nanogel made of pullulan with vinyl methacrylate and more hydrophobic chains grafted was affected by the ionic strength and urea, nanogel made of pullulan with hydroxyethyl methacrylate and fewer hydrophobic chains grafted remained stable.

  12. Surfactantless synthesis and textural properties of self-assembled mesoporous SnO{sub 2}

    Energy Technology Data Exchange (ETDEWEB)

    Velasquez, Celso [Departamento de QuImica, Universidad Autonoma Metropolitana-Iztapalapa, PO Box 55-534, Mexico, DF 09340 (Mexico); Ojeda, MarIa Luisa [Instituto de QuImica, UNAM, Circuito Exterior, Ciudad Universitaria, CP 04510, Mexico, DF (Mexico); Campero, Antonio [Departamento de QuImica, Universidad Autonoma Metropolitana-Iztapalapa, PO Box 55-534, Mexico, DF 09340 (Mexico); Esparza, Juan Marcos [Departamento de QuImica, Universidad Autonoma Metropolitana-Iztapalapa, PO Box 55-534, Mexico, DF 09340 (Mexico); Rojas, Fernando [Departamento de QuImica, Universidad Autonoma Metropolitana-Iztapalapa, PO Box 55-534, Mexico, DF 09340 (Mexico)


    Ordered surfactantless self-assembled, mesoporous SnO{sub 2} adsorbents, consisting of tubular voids of nanometric sizes, are prepared by the sol-gel processing of tin (IV) tetra-tert-amyloxide, Sn(OAm{sup t}){sub 4}, whose molecules have been previously chelated with acetylacetone in the absence of water, to modulate their reactivity and to promote an incipient self-assembling of -O-Sn-O oligomeric species; ultimately, the necessary amount of water to induce the hydrolysis-condensation reactions is added to this aged sol, then producing tubular pore templates within the SnO{sub 2} xerogel network. A collection of mesoporous SnO{sub 2} xerogels of assorted structural properties has been obtained after calcination in air of precursory gels proceeding from an aged mixture of Sn(OAm{sup t}){sub 4} and acetylacetone at temperatures in the range 200-1000 deg. C. N{sub 2} sorption isotherms measured on these SnO{sub 2} solids evidence mesoporous structures of diverse textural characteristics (i.e. pore widths of 3-50 nm and surface areas of 10-140 m{sup 2} g{sup -1}) in which voids virtually behave as if they are independent cylindrical pores during capillary condensation and evaporation.

  13. DNA synthesis and microtubule assembly-related events in fertilized Paracentrotus lividus eggs: reversible inhibition by 10 mM procaine. (United States)

    Raymond, M N; Foucault, G; Coffe, G; Pudles, J


    This report describes the effects of 10 mM procaine on microtubule assembly and on DNA synthesis, as followed by [3H]colchicine binding assays and [3H]thymidine incorporation respectively, in fertilized Paracentrotus lividus eggs. In the absence of microtubule assembly inhibitors, about 25% of the total egg tubulin is submitted to two cycles of polymerization prior to the first cell division, this polymerization process precedes DNA synthesis. If the zygotes are treated with 10 mM procaine in the course of the cell cycle, tubulin polymerization is inhibited or microtubules are disassembled. DNA synthesis is inhibited when procaine treatment is performed 10 min, before the initiation of the S-period. However, when the drug is applied in the course of this synthetic period, the process is normally accomplished, but the next S-period becomes inhibited. Moreover, procaine treatment increases the cytoplasmic pH of the fertilized eggs by about 0.6 to 0.8 pH units. This pH increase precedes microtubule disassembly and inhibition of DNA synthesis. Washing out the drug induces a decrease of the intracellular pH which returns to about the same value as that of the fertilized egg controls. This pH change is then followed by the reinitiation of microtubule assembly, DNA synthesis and cell division. Our results show that the inhibition of both tubulin polymerization and DNA synthesis in fertilized eggs treated with 10 mM procaine, appears to be related to the drug-induced increase in cytoplasmic pH.


    Directory of Open Access Journals (Sweden)

    Shekhawat G. S.


    Full Text Available In recent years role of Heme oxygenase (HO has been considered in nearly all living system including plants, animals and other organisms. The common role of heme oxygenase is the degradation of heme, although there is a diversity of additional role of HO in organisms including iron acquisition, cellular signaling, defense against stress and biosynthesis during metabolism. Likewise, the function of HO is to provide cofactors for the photosynthetic apparatus in cyanobacteria. Heme concentration is variable in different plant species and found maximum in leguminous plant root nodules. Moreover HO has diverse isoforms in plant and animal systems. The review addressed important function of HO and focused on its functional diversity.

  15. Synthesis and characterization of polystyrene coated iron oxide nanoparticles and asymmetric assemblies by phase inversion

    KAUST Repository

    Xie, Yihui


    Films with a gradient concentration of magnetic iron oxide nanoparticles are reported, based on a phase inversion membrane process. Nanoparticles with ∼13 nm diameter were prepared by coprecipitation in aqueous solution and stabilized by oleic acid. They were further functionalized by ATRP leading to grafted polystyrene brush. The final nanoparticles of 33 nm diameter were characterized by TGA, FTIR spectroscopy, GPC, transmission electron microscopy, and dynanmic light scattering. Asymmetric porous nanoparticle assemblies were then prepared by solution casting and immersion in water. The nanocomposite film production with functionalized nanoparticles is fast and technically scalable. The morphologies of films were characterized by scanning electron microscopy and atomic force microscopy, demonstrating the presence of sponge-like structures and finger-like cavities when 50 and 13 wt % casting solutions were, respectively, used. The magnetic properties were evaluated using vibrating sample magnetometer.

  16. Self-assembled hexanuclear organometallic cages: synthesis, characterization, and host-guest properties. (United States)

    Han, Ying-Feng; Li, Hao; Zheng, Zhi-Fang; Jin, Guo-Xin


    A series of iridium- and rhodium-based hexanuclear organometallic cages containing 2,5-dichloro-3,6-dihydroxy-1,4-benzoquinone, 9,10-dihydroxy-1,4-anthraquinone, and 6,11-dihydroxynaphthacene-5,12-dione ligands were synthesized from the self-assembly of the corresponding molecular "clips" and 2,4,6-tri(4-pyridyl)-1,3,5-triazine ligands in good yields. These organometallic cages can form inclusion systems with a wide variety of π-donor substrates, including coronene, pyrene, [Pt(acac)(2)], and hexamethoxytriphenylene. The 1:1 complexation of the resulting supramolecular assemblies was confirmed by (1)H NMR spectroscopy. Large complexation shifts (Δδ>1 ppm) were observed in the (1)H NMR spectra of guests in the presence of cage [Cp*(6)M(6)(μ-DHNA)(3)(tpt)(2)](OTf)(6) (6a; M=Ir, tpt=2,4,6-tri(4-pyridyl)-1,3,5-triazine). The formation of discrete 1:1 donor-acceptor complexes, pyrene⊂6b (M=Rh), coronene⊂6a, coronene⊂6b, and [Pt(acac)(2)]⊂6a was confirmed by their single-crystal X-ray analyses. In these systems, the most important driving force for the formation of guest-host complexes is clearly the donor-acceptor π···π stacking interaction, including charge-transfer interactions between the electron-donating and electron-accepting aromatic components. These structures provide compelling evidence for the existence of strong attractive forces between the electron-deficient triazine core and electron-rich guest. The results presented here may provide useful guidance for designing artificial receptors for functional biomolecules.

  17. Structural mechanisms of nonplanar hemes in proteins

    Energy Technology Data Exchange (ETDEWEB)

    Shelnutt, J.A.


    The objective is to assess the occurrence of nonplanar distortions of hemes and other tetrapyrroles in proteins and to determine the biological function of these distortions. Recently, these distortions were found by us to be conserved among proteins belonging to a functional class. Conservation of the conformation of the heme indicates a possible functional role. Researchers have suggested possible mechanisms by which heme distortions might influence biological properties; however, no heme distortion has yet been shown conclusively to participate in a structural mechanism of hemoprotein function. The specific aims of the proposed work are: (1) to characterize and quantify the distortions of the hemes in all of the more than 300 hemoprotein X-ray crystal structures in terms of displacements along the lowest-frequency normal coordinates, (2) to determine the structural features of the protein component that generate and control these nonplanar distortions by using spectroscopic studies and molecular-mechanics calculations for the native proteins, their mutants and heme-peptide fragments, and model porphyrins, (3) to determine spectroscopic markers for the various types of distortion, and, finally, (4) to discover the functional significance of the nonplanar distortions by correlating function with porphyrin conformation for proteins and model porphyrins.

  18. Crystal structure of dimeric heme oxygenase-2 from Synechocystis sp. PCC 6803 in complex with heme. (United States)

    Sugishima, Masakazu; Hagiwara, Yoshinori; Zhang, Xuhong; Yoshida, Tadashi; Migita, Catharina T; Fukuyama, Keiichi


    Phycobiliproteins, light-harvesting proteins in cyanobacteria, red algae, and cryptophytes, contain phycobilin pigments. Phycobilins are synthesized from biliverdin, which is produced by the oxidative cleavage of the heme porphyrin ring catalyzed by heme oxygenase (HO). Two paralogs of ho (ho1 and ho2) have been identified in the genome of the cyanobacterium, Synechocystis sp. PCC 6803. The recombinant proteins of both paralogs (Syn HO-1 and Syn HO-2) possess in vitro heme degradation activity. We have determined the crystal structures of Syn HO-2 in complex with heme (heme-Syn HO-2) and its reduced and NO bound forms. The heme-Syn HO-2 crystal was a nonmerohedral twin, and detwinned diffraction data were used to refine the structure. Although heme-Syn HO-2 shares common folding with other HOs, the C-terminal segment is ordered and turns back to the heme-binding side. Gel-filtration chromatography analysis and molecular packing in the crystal indicate that heme-Syn HO-2 forms a homodimer, in which the C-terminal ordered segments interact with each other. Because Syn HO-2 is a monomer in the apo state, the dimeric interaction may aid in the selection of the reducing partner but likely does not interfere with heme binding. The heme iron is coordinated by a water molecule in the ferric form, but the distal water is absent in the ferrous form. In all of the Syn HO-2 structures, several water molecules form a hydrogen-bond network at the distal hemepocket, which is involved in HO activity. Upon NO binding, the side-chain conformation of Tyr 156 changes. Tyr 156 is located at the hydrophobic cluster, which interrupts the possible H(+) pathway from the molecular surface to the hemepocket. Thus, Tyr 156 may function as a H(+) shuttle by changing conformation.

  19. Identification of the receptor scavenging hemopexin-heme complexes

    DEFF Research Database (Denmark)

    Hvidberg, Vibeke; Maniecki, Maciej B; Jacobsen, Christian;


    and is suggested to facilitate cellular heme metabolism. Using a ligand-affinity approach, we purified the human hemopexin-heme receptor and identified it as the low-density lipoprotein receptor-related protein (LRP)/CD91, a receptor expressed in several cell types including macrophages, hepatocytes, neurons......, and syncytiotrophoblasts. Binding experiments, including Biacore analysis, showed that hemopexin-heme complex formation elicits the high receptor affinity. Uptake studies of radio-labeled hemopexin-heme complex in LRP/CD91-expressing COS cells and confocal microscopy of the cellular processing of fluorescent hemopexin......-heme complex established the ability of LRP/CD91 to mediate hemopexin-heme internalization resulting in cellular heme uptake and lysosomal hemopexin degradation. Uptake of hemopexin-heme complex induced LRP/CD91-dependent heme-oxygenase 1 mRNA transcription in cultured monocytes. In conclusion, hemopexin...

  20. Heme electron transfer in peroxidases: the propionate e-pathway. (United States)

    Guallar, Victor


    Computational modeling offers a new insight about the electron transfer pathway in heme peroxidases. Available crystal structures have revealed an intriguing arrangement of the heme propionate side chains in heme-heme and heme-substrate complexes. By means of mixed quantum mechanical/molecular mechanics calculations, we study the involvement of these propionate groups into the substrate oxidation in ascorbate peroxidase and into the heme to heme electron transfer in bacterial cytochrome c peroxidase. By selectively turning on/off different quantum regions, we obtain the electron transfer pathway which directly involves the porphyrin ring and the heme propionates. Furthermore, in ascorbate peroxidase the presence of the substrate appears to be crucial for the activation of the electron transfer channel. The results might represent a general motif for electron transfer from/to the heme group and change our view for the propionate side chains as simple electrostatic binding anchors. We name the new mechanism "the propionate e-pathway".

  1. NikA binds heme: a new role for an Escherichia coli periplasmic nickel-binding protein. (United States)

    Shepherd, Mark; Heath, Mathew D; Poole, Robert K


    NikA is a periplasmic binding protein involved in nickel uptake in Escherichia coli. NikA was identified as a heme-binding protein in the periplasm of anaerobically grown cells overexpressing CydDC, an ABC transporter that exports reductant to the periplasm. CydDC-overexpressing cells accumulate a heme biosynthesis-derived pigment, P-574. For further biochemical and spectroscopic analysis, unliganded NikA was overexpressed and purified. NikA was found to comigrate with both hemin and protoporphyrin IX during gel filtration. Furthermore, tryptophan fluorescence quenching titrations demonstrated that both hemin and protoporphyrin IX bind to NikA with similar affinity. The binding affinity of NikA for these pigments (Kd approximately 0.5 microM) was unaltered in the presence and absence of saturating concentrations of nickel, suggesting that these tetrapyrroles bind to NikA in a manner independent of nickel. To test the hypothesis that NikA is required for periplasmic heme protein assembly, the effects of a nikA mutation (nikA::Tn5, Km(R) insertion) on accumulation of P-574 by CydDC-overexpressing cells was assessed. This mutation significantly lowered P-574 levels, implying that NikA may be involved in P-574 production. Thus, in the reducing environment of the periplasm, NikA may serve as a heme chaperone as well as a periplasmic nickel-binding protein. The docking of heme onto NikA was modeled using the published crystal structure; many of the predicted complexes exhibit a heme-binding cleft remote from the nickel-binding site, which is consistent with the independent binding of nickel and heme. This work has implications for the incorporation of heme into b- and c-type cytochromes.

  2. Synthesis, physical properties and self-assembly behavior of azole-fused pyrene derivatives (United States)

    Xiao, Jinchong; Xiao, Xuyu; Zhao, Yanlei; Wu, Bo; Liu, Zhenying; Zhang, Xuemin; Wang, Sujuan; Zhao, Xiaohui; Liu, Lei; Jiang, Li


    A novel selenadiazole-fused pyrene derivative PySe was successfully synthesized and characterized. Its single structure is almost planar and adopts a sandwich-herringbone packing model. The self-assembly behaviors based on compound PySe and its analogue thiadiazole-fused pyrene derivative PyS were studied in detail and the as-formed nanostructures were fully characterized by means of UV-vis absorption, emission spectra, X-ray diffraction, field emission SEM and TEM. We attribute the bathochromic shift absorption and emission spectra of PyS and PySe in aqueous solution to the formation of J-type aggregation. In addition, our investigation demonstrated that the shape and size of the as-prepared nanostructures could be tuned by different chalcogen analogues and the volume ratio of water to organic solvent.A novel selenadiazole-fused pyrene derivative PySe was successfully synthesized and characterized. Its single structure is almost planar and adopts a sandwich-herringbone packing model. The self-assembly behaviors based on compound PySe and its analogue thiadiazole-fused pyrene derivative PyS were studied in detail and the as-formed nanostructures were fully characterized by means of UV-vis absorption, emission spectra, X-ray diffraction, field emission SEM and TEM. We attribute the bathochromic shift absorption and emission spectra of PyS and PySe in aqueous solution to the formation of J-type aggregation. In addition, our investigation demonstrated that the shape and size of the as-prepared nanostructures could be tuned by different chalcogen analogues and the volume ratio of water to organic solvent. Electronic supplementary information (ESI) available: TGA analysis, spectra characterization data for compound 1, 2, 3 and X-ray crystallographic data for compound PySe (2, CIF). CCDC 917821. For ESI and crystallographic data in CIF or other electronic format see DOI: 10.1039/c3nr00523b

  3. Superfast assembly and synthesis of gold nanostructures using nanosecond low-temperature compression via magnetic pulsed power (United States)

    Li, Binsong; Bian, Kaifu; Lane, J. Matthew D.; Salerno, K. Michael; Grest, Gary S.; Ao, Tommy; Hickman, Randy; Wise, Jack; Wang, Zhongwu; Fan, Hongyou


    Gold nanostructured materials exhibit important size- and shape-dependent properties that enable a wide variety of applications in photocatalysis, nanoelectronics and phototherapy. Here we show the use of superfast dynamic compression to synthesize extended gold nanostructures, such as nanorods, nanowires and nanosheets, with nanosecond coalescence times. Using a pulsed power generator, we ramp compress spherical gold nanoparticle arrays to pressures of tens of GPa, demonstrating pressure-driven assembly beyond the quasi-static regime of the diamond anvil cell. Our dynamic magnetic ramp compression approach produces smooth, shockless (that is, isentropic) one-dimensional loading with low-temperature states suitable for nanostructure synthesis. Transmission electron microscopy clearly establishes that various gold architectures are formed through compressive mesoscale coalescences of spherical gold nanoparticles, which is further confirmed by in-situ synchrotron X-ray studies and large-scale simulation. This nanofabrication approach applies magnetically driven uniaxial ramp compression to mimic established embossing and imprinting processes, but at ultra-short (nanosecond) timescales. PMID:28300067

  4. Facile Synthesis of Gold-nanoparticles-decorated Polymer Assemblies and Core-Shell Gold Nanoparticles Using Pluronic Block Copolymers

    Institute of Scientific and Technical Information of China (English)

    SHOU Qing-hui; GUO Chen; GAO Hong-shuai; ZHOU Hua-cong; LIU Chun-zhao; LIU Hui-zhou


    Abstract:Synthesis of gold nanoparticles (AuNPs) and Pluronic triblock copolymer composite in aqueous medium was studied.Gold-polymer nanocomposite with different structures was fabricated by tailoring the molar ratio of gold precursors to Pluronic P123 molecules or pH value of the P123 solution.When a lower volume ratio of [AuCl4-]/[P123] (0.05) was employed at pH 11.1,a nanostructure similar to plum pudding was obtained.AuNPs with an average diameter of 13.1 nm were embedded in Pluronic assemblies,and each one held about 21 single gold nanoparticles.When [AuCl4-]/[P123] was increased to 0.1,core-shell structure was obtained if the pH value was in the range of 10.6~11.6,while gold polyhedra were fabricated when pH value was 8.1.Typical core-shell AuNPs had an average diameter of 9.6 nm with a narrow size distribution,while gold polyhedras with a mean diameter of 12.8 nm was obtained.The specific morphologies of the resultant nanocomposite were presumably obtained due to the synergistic interaction among the reactants.

  5. Magnetic self-assembly for the synthesis of magnetically exchange coupled MnBi/Fe–Co composites

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Xia [Department of Chemical and Biological Engineering and MINT Center, The University of Alabama, Tuscaloosa, AL 35487 (United States); Hong, Yang-Ki, E-mail: [Department of Electrical and Computer Engineering and MINT Center, The University of Alabama, Tuscaloosa, AL 35487 (United States); Park, Jihoon; Lee, Woncheol [Department of Electrical and Computer Engineering and MINT Center, The University of Alabama, Tuscaloosa, AL 35487 (United States); Lane, Alan M. [Department of Chemical and Biological Engineering and MINT Center, The University of Alabama, Tuscaloosa, AL 35487 (United States); Cui, Jun [Energy and Environment Directorate, Pacific Northwestern National Laboratory, Richland, WA 99354 (United States)


    Exchange coupled hard/soft MnBi/Fe–Co core/shell structured composites were synthesized using a magnetic self-assembly process. MnBi particles were prepared by arc-melting, and Fe–Co nanoparticles were synthesized by an oleic acid assisted chemical reduction method. Grinding a mixture of micron-sized MnBi and Fe–Co nanoparticles in hexane resulted in MnBi/Fe–Co core/shell structured composites. The MnBi/Fe–Co (95/5 wt%) composites showed smooth magnetic hysteresis loops, enhanced remanent magnetization, and positive values in the ΔM curve, indicating exchange coupling between MnBi and Fe–Co particles. - Graphical abstract: Both MnBi and Fe–Co particles were dispersed in hexane for grinding. Because of the oleic acid used during the Fe–Co nanoparticle synthesis, they could be well dispersed in hexane. During the grinding, the size of MnBi particles was decreased, hexane was evaporated, and the Fe–Co nanoparticles were concentrated in the solvent and magnetically attracted by MnBi particles, forming a core/shell structure. - Highlights: • Exchange coupled MnBi/Fe–Co composites are synthesized through magnetic selfassembly. • Magnetic exchange coupling is demonstrated by smooth magnetic hysteresis loops, enhanced remanent magnetization, and dominant positive peak in the ΔM curve. • The experimental results in magnetic properties are close to the theoretical calculation results.

  6. Lanthanide-directed synthesis of luminescent self-assembly supramolecular structures and mechanically bonded systems from acyclic coordinating organic ligands. (United States)

    Barry, Dawn E; Caffrey, David F; Gunnlaugsson, Thorfinnur


    Herein some examples of the use of lanthanide ions (f-metal ions) to direct the synthesis of luminescent self-assembly systems (architectures) will be discussed. This area of lanthanide supramolecular chemistry is fast growing, thanks to the unique physical (magnetic and luminescent) and coordination properties of the lanthanides, which are often transferred to the resulting supermolecule. The emphasis herein will be on systems that are luminescent, and hence, generated by using either visibly emitting ions (such as Eu(III), Tb(III) and Sm(III)) or near infrared emitting ions (like Nd(III), Yb(III) and Er(III)), formed through the use of templating chemistry, by employing structurally simple ligands, possessing oxygen and nitrogen coordinating moieties. As the lanthanides have high coordination requirements, their use often allows for the formation of coordination compounds and supramolecular systems such as bundles, grids, helicates and interlocked molecules that are not synthetically accessible through the use of other commonly used templating ions such as transition metal ions. Hence, the use of the rare-earth metal ions can lead to the formation of unique and stable species in both solution and in the solid state, as well as functional and responsive structures.

  7. Size- and shape-controlled synthesis of hexagonal bipyramidal crystals and hollow self-assembled Al-MOF spheres

    KAUST Repository

    Sarawade, Pradip


    We report an efficient protocol for the synthesis of monodisperse crystals of an aluminum (Al)-based metal organic framework (MOF) while obtaining excellent control over the size and shape solely by tuning of the reaction parameters without the use of a template or structure-directing agent. The size of the hexagonal crystals of the Al-MOF can be selectively varied from 100 nm to 2000 nm by simply changing the reaction time and temperature via its nucleation-growth mechanism. We also report a self-assembly phenomenon, observed for the first time in case of Al-MOF, whereby hollow spheres of Al-MOF were formed by the spontaneous organization of triangular sheet building blocks. These MOFs showed broad hysteresis loops during the CO2 capture, indicating that the adsorbed CO2 is not immediately desorbed upon decreasing the external pressure and is instead confined within the framework, which allows for the capture and subsequent selective trapping of CO2 from gaseous mixtures. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. A spray-drying strategy for synthesis of nanoscale metal-organic frameworks and their assembly into hollow superstructures. (United States)

    Carné-Sánchez, Arnau; Imaz, Inhar; Cano-Sarabia, Mary; Maspoch, Daniel


    Metal-organic frameworks (MOFs) are among the most attractive porous materials known today. Their miniaturization to the nanoscale--into nanoMOFs--is expected to serve myriad applications from drug delivery to membranes, to open up novel avenues to more traditional storage and catalysis applications, and to enable the creation of sophisticated superstructures. Here, we report the use of spray-drying as a versatile methodology to assemble nanoMOFs, yielding spherical hollow superstructures with diameters smaller than 5 µm. This strategy conceptually mimics the emulsions used by chemists to confine the synthesis of materials, but does not require secondary immiscible solvents or surfactants. We demonstrate that the resulting spherical, hollow superstructures can be processed into stable colloids, whose disassembly by sonication affords discrete, homogeneous nanoMOFs. This spray-drying strategy enables the construction of multicomponent MOF superstructures, and the encapsulation of guest species within these superstructures. We anticipate that this will provide new routes to capsules, reactors and composite materials.

  9. Synthesis and cell localization of self-assembled dinuclear lanthanide bioprobes. (United States)

    Chauvin, Anne-Sophie; Thomas, Frédéric; Song, Bo; Vandevyver, Caroline D B; Bünzli, Jean-Claude G


    Lanthanide bioprobes and bioconjugates are ideal luminescent stains in view of their low propensity to photobleaching, sharp emission lines and long excited state lifetimes permitting time-resolved detection for enhanced sensitivity. In this paper, we expand our previous work which demonstrated that self-assembled dinuclear triple-stranded helicates [Ln2(L(C2X))3] behave as excellent cell and tissue labels in immunocytochemical and immunohistochemical assays. The synthetic strategy of the hexadentate ditopic ligands incorporating dipicolinic acid, benzimidazole units and polyoxyethylene pendants is revisited in order to provide a more straightforward route and to give access to further functionalization of the polyoxyethylene arms by incorporating a terminal function X. Formation of the helicates [Ln2(L(C2X))3] (X=COOH, CH2OH, COEt, NH2, phthalimide) is ascertained by several experimental techniques and their stability tested against diethylenetriaminepentaacetate. Their photophysical properties (quantum yield, lifetime, radiative lifetime and sensitization efficiency) are presented and compared with those of the parent helicates [Ln2(L(C2))3]. Finally, the cellular uptake of five Eu(III) helicates is monitored by time-resolved luminescence microscopy and their localization in HeLa cells established by co-staining experiments.

  10. Shape-controlled synthesis of gold nanoplates and their self-assembly by repulsive electrostatic interactions. (United States)

    Bi, Liyan; Rao, Yanying; Sun, Qian; Li, Danyang; Cheng, Yuan; Dong, Jian; Qian, Weiping


    This study focuses on understanding the growth and control of the gold nanoplates by seed-mediated growth approach. These monodispersive size-controlled gold nanoplates have the average thickness of 8-10 nm and average size tunable from 70 to 150 nm, exhibiting strong surface plasmon absorption in the near infrared (NIR) region. For the gold nanoplates formation, the seeds serve as nucleation sites, ascorbic acid (AA) serves as a new reductant to reduce hydrogen tetrachloroaurate (HAuCl4), surface activity system cetyltrimethylammonium bromide (CTAB) and potassium iodide (KI) are critical factors. X-ray diffraction (XRD) and selected-area electron diffraction (SAED) analyses reveal that gold nanoplates with the (111) lattice plane as the basal plane are single crystals. CTAB are absorbed on the surface of the (111) lattice plane of the single crystals, accounting for self-assembled monolayer and head-to-head arrays. The two arrays have been shown to serve as effective surface-enhanced Raman scattering substrates using Niel blue A (NBA) sulfate as Raman report molecule.

  11. Synthesis, characterization, and assembly of beta-In2S3 nanoparticles. (United States)

    Vigneashwari, B; Dash, S; Tyagi, A K; Parameswaran, P; Ravichandran, V; Sunthathiraraj, S Austin


    Semiconductor nanoparticles of Indium Sulphide were synthesized by a hydrothermal method using InCl3 and Na2S. Powder X-ray Diffraction analysis confirmed that the product obtained was nanocrystals of single-phase beta-In2S3. The crystallite size distribution was obtained from the diffraction profile and the average size was approximately 5 nm. The compositional analyses performed on the as-prepared powder showed that the material was devoid of any impurity with an In:S ratio very close to 2:3. A colloid of very fine In2S3 particles was obtained from the as-prepared powder by suspending them in acetonitrile. The optical absorption of this colloid showed evidence of strong quantum confinement of excitons and as a result the particles yielded intense photoluminescence in the violet-blue region. These colloidal particles were then electrophoretically driven on to a transparent conducting substrate to assemble into a nanostructure. A Grazing Incidence X-ray Diffraction analysis of the deposited layer revealed that the preferred orientation noticed in the native powder was removed in the deposit. The surface morphology of the deposit studied using SEM and AFM displayed an inherent ordering behaviour in the clusters organized into a two-dimensional film. The locus of the cluster lines tend to form closed circles, at the nanoscopic as well as microscopic scales, indicative of certain strong neighborhood correlations. Such structures may be expected to exhibit novel correlated properties also.

  12. Synthesis of Self-assembled Noble Metal Nanoparticle Chains Using Amyloid Fibrils of Lysozyme as Templates

    Directory of Open Access Journals (Sweden)

    Ziming Xu


    Full Text Available We reported a facile method for preparing self-assembled noble metal nanoparticle chains by using lysozyme amyloid fibrils as a biotemplate in an aqueous environ‐ ment. The nanoparticle chains of gold (AuNPCs, palladi‐ um (PdNPCs, platinum (PtNPCs and rhodium (RhNPCs, which are lysozyme fibrils coated by gold, palladium, platinum and rhodium nanoparticles, can be fabricated by simply reducing the corresponding metal salt precursors using NaBH4. Under the same molar ratio between salt precursors and fibrils, two types of morphologies of high- yield AuNPCs (thin- and thick- AuNPCs were synthesized as a result of adjusting the fibrosis time and temperature in the final stage. Abundant PdNPCs with a length of several micrometres intertwisted with each other to form PdNPC networks. The growth of RhNPCs started from the inner surface of the fibrils and gradually spread to the whole fibre as superabundant rhodium nanoparticles (RhNPs bound to the fibrils. Finally, PtNPCs at different growing periods were presented. The nanostructures were investigated by transmission electron microscope, UV-visible spectrosco‐ py, fluorescence spectroscopy, energy-dispersive X-ray spectroscopy and atomic force microscope.

  13. Synthesis, characterization and self-assembly of well-defined linear heptablock quaterpolymers

    KAUST Repository

    Ntaras, Christos


    Two well-defined heptablock quaterpolymers of the ABCDCBA type [Α: polystyrene (PS), B: poly(butadiene) with ∼90% 1,4-microstructure (PB1,4), C: poly(isoprene) with ∼55% 3,4-microstructure (PI3,4) and D: poly(dimethylsiloxane) (PDMS)] were synthesized by combining anionic polymerization high vacuum techniques and hydrosilylation/chlorosilane chemistry. All intermediates and final products were characterized by size exclusion chromatography, membrane osmometry, and proton nuclear magnetic resonance spectroscopy. Fourier transform infrared spectroscopy was used to further verify the chemical modification reaction of the difunctional PDMS. The self-assembly in bulk of these novel heptablock quarterpolymers, studied by transmission electron microscopy and small angle X-ray scattering, revealed 3-phase 4-layer alternating lamellae morphology of PS, PB1,4, and mixed PI3,4/PDMS domains. Differential scanning calorimetry was used to further confirm the miscibility of PI3,4 and PDMS blocks. It is the first time that PDMS is the central segment in such multiblock polymers (≥3 chemically different blocks). © 2016 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2016, 54, 1443–1449. © 2016 Wiley Periodicals, Inc.

  14. Fluorine-Containing ABC Linear Triblock Terpolymers: Synthesis and Self-assembly in Solution

    Energy Technology Data Exchange (ETDEWEB)

    He, Lihong [ORNL; Hinestrosa Salazar, Juan P [ORNL; Pickel, Joseph M [ORNL; Kilbey, II, S Michael [ORNL; Mays, Jimmy [ORNL; Zhang, Shanju [Georgia Institute of Technology; Bucknall, David G. [Georgia Institute of Technology; Hong, Kunlun [ORNL


    In this paper a fluorine-containing monomer, 2-fluroroethyl methacrylate (2FEMA) was used to synthesize the linear triblock terpolymer poly(n-butyl methacrylate)-poly(methyl methacrylate)-poly(2-fluoroethyl methacrylate) (PnBMA-PMMA-P2FEMA). A kinetic study of the homopolymerization of 2FEMA by reversible addition-fragmentation chain transfer (RAFT) polymerization showed that it demonstrates living character and produces well defined polymers with reasonably narrow polydispersities (~1.30). Triblock terpolymers were prepared sequentially using a purified Macro-CTA at 70 oC, resulting in final terpolymers with high Dp for each block (>150) and with polydispersities between 1.6 and 2.1. The structure and molecular weights of the resultant PnBMA-PMMA-P2FEMA triblock terpolymers were characterized via 1H NMR, 19F NMR, and gel permeation chromatography (GPC). Self-assembly of these polymers was carried out in a selective solvent and the micellar aggregates (MAs) thereby formed were analyzed using scanning electron microscopy (SEM) and dynamic light scattering (DLS). It was confirmed from SEM that these copolymers could directly self-organize into large compound micelles in tetrahydrofuran/methanol with different diameters, depending on polymer composition.

  15. Colloid electrostatic self-assembly synthesis of SnO{sub 2}/graphene nanocomposite for supercapacitors

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Yankun; Liu, Yushan; Zhang, Jianmin, E-mail: [Zhengzhou University, College of Chemistry and Molecular Engineering (China)


    In this paper, a simple and fast colloid electrostatic self-assembly method was adopted to prepare the SnO{sub 2}/graphene nanocomposite (SGNC). The crystal structure, chemical composition, and porous property of composite were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, Raman microscopy, X-ray photoelectron spectroscopy (XPS), and N{sub 2} adsorption–desorption experiments. The morphology analyses showed that the SnO{sub 2} nanoparticles about 5 nm were distributed homogenously on the reduced graphene oxide (rGO) sheets surface. The electrochemical performance measurements exhibited that SGNC possessed the specific capacitance of 347.3 F g{sup −1} at a scan rate of 5 mV s{sup −1} in 1 M Na{sub 2}SO{sub 4} electrolyte solution. Furthermore, this material also showed excellent cycling stability, and the specific capacitance still retained 90 % after 3000 cycles. These results indicate that the SGNC is a promising electrode material for high-performance supercapacitors.

  16. Synthesis of self-assembled Ge nano crystals employing reactive RF sputtering

    Energy Technology Data Exchange (ETDEWEB)

    Hernandez H, A. [Universidad Autonoma del Estado de Hidalgo, Escuela Superior de Apan, Calle Ejido de Chimalpa Tlalayote s/n, Col. Chimalpa, Apan, Hidalgo (Mexico); Hernandez H, L. A. [IPN, Escuela Superior de Fisica y Matematicas, San Pedro Zacatenco, 07730 Ciudad de Mexico (Mexico); Monroy, B. M.; Santana R, G. [UNAM, Instituto de Investigaciones en Materiales, Apdo. Postal 70-360, 04510 Ciudad de Mexico (Mexico); Santoyo S, J.; Gallardo H, S. [IPN, Centro de Investigacion y de Estudios Avanzados, Departamento de Fisica, Apdo. Postal 14740, 07300 Ciudad de Mexico (Mexico); Marquez H, A. [Universidad de Guanajuato, Campus Irapuato-Salamanca, Departamento de Ingenieria Agricola, Km. 9 Carretera Irapuato-Silao, 36500 Irapuato, Guanajuato (Mexico); Mani G, P. G.; Melendez L, M. [Universidad Autonoma de Ciudad Juarez, Instituto de Ingenieria y Tecnologia, Departamento de Fisica y Matematicas, 32310 Ciudad Juarez, Chihuahua (Mexico)


    This work presents the results of a simple methodology able to control crystal size, dispersion and spatial distribution of germanium nano crystals (Ge-NCs). It takes advantage of a self-assembled process taken place during the deposit of the system SiO{sub 2}/Ge/SiO{sub 2} by reactive RF sputtering. Nanoparticles formation is controlled mainly by the roughness of the first SiO{sub 2} layer buy the ulterior interaction of the interlayer with the top layer also play a role. Structural quality of germanium nano crystals increases with roughness and the interlayer thickness. The tetragonal phase of germanium is produced and its crystallographic quality improves with interlayer thickness and oxygen partial pressure. Room temperature photoluminescence emission without a post growth thermal annealing process indicates that our methodology produces a low density of non-radiative traps. The surface topography of SiO{sub 2} reference samples was carried out by atomic force microscopy. The crystallographic properties of the samples were studied by grazing incidence X-ray diffraction at 1.5 degrees carried out in a Siemens D-5000 system employing the Cu Kα wavelength. (Author)

  17. Synthesis, characterization and anticorrosion potentials of chitosan-g-PEG assembled on silver nanoparticles. (United States)

    Hefni, Hassan H H; Azzam, Eid M; Badr, Emad A; Hussein, M; Tawfik, Salah M


    Chitosan (Ch) grafted with poly(ethylene glycol) (Ch-g-mPEG) were synthesized using mPEG with molecular weights 2000 g/mol. The synthesized Ch-g-mPEG was characterized using gel permeation chromatography (GPC), Fourier transform infrared spectroscopy (FTIR), proton nuclear magnetic resonance (1H NMR), and X-ray diffraction (XRD) techniques. Ch-g-mPEG silver nanoparticles has been synthesized and characterized by high-resolution transmission electron microscopy (HRTEM) and energy dispersive analysis of X-rays (EDAX). The synthesized Ch-g-mPEG and its nanostructure were examined as corrosion inhibitors for carbon steel in 1M HCl solution using potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) techniques. The results revealed that the inhibition efficiency obtained by Ch-g-mPEG self-assembled on silver nanoparticles is greater than that obtained by Ch-g-mPEG only. Potentiodynamic polarization results reveal that the synthesized compound could be classified as mixed-type corrosion inhibitors with predominant control of the cathodic reaction. The results of EIS indicate that the both charge transfer resistance and inhibition efficiency tend to increase by increasing the inhibitor concentration.

  18. Heme-containing dioxygenases involved in tryptophan oxidation. (United States)

    Millett, Elizabeth S; Efimov, Igor; Basran, Jaswir; Handa, Sandeep; Mowat, Christopher G; Raven, Emma Lloyd


    Heme iron is often used in biology for activation of oxygen. The mechanisms of oxygen activation by heme-containing monooxygenases (the cytochrome P450s) are well known, and involve formation of a Compound I species, but information on the heme-containing dioxygenase enzymes involved in tryptophan oxidation lags far behind. In this review, we gather together information emerging recently from structural, mechanistic, spectroscopic, and computational approaches on the heme dioxygenase enzymes involved in tryptophan oxidation. We explore the subtleties that differentiate various heme enzymes from each other, and use this to piece together a developing picture for oxygen activation in this particular class of heme-containing dioxygenases.

  19. The non-canonical functions of the heme oxygenases. (United States)

    Vanella, Luca; Barbagallo, Ignazio; Tibullo, Daniele; Forte, Stefano; Zappalà, Agata; Li Volti, Giovanni


    Heme oxygenase (HO) isoforms catalyze the conversion of heme to carbon monoxide (CO) and biliverdin with a concurrent release of iron, which can drive the synthesis of ferritin for iron sequestration. Most of the studies so far were directed at evaluating the protective effect of these enzymes because of their ability to generate antioxidant and antiapoptotic molecules such as CO and bilirubin. Recent evidences are suggesting that HO may possess other important physiological functions, which are not related to its enzymatic activity and for which we would like to introduce for the first time the term "non canonical functions". Recent evidence suggest that both HO isoforms may form protein-protein interactions (i.e. cytochrome P450, adiponectin, CD91) thus serving as chaperone-like protein. In addition, truncated HO-1 isoform was localized in the nuclear compartment under certain experimental conditions (i.e. excitotoxicity, hypoxia) regulating the activity of important nuclear transcription factors (i.e. Nrf2) and DNA repair. In the present review, we discuss three potential signaling mechanisms that we refer to as the non-canonical functions of the HO isoforms: protein-protein interaction, intracellular compartmentalization, and extracellular secretion. The aim of the present review is to describe each of this mechanism and all the aspects warranting additional studies in order to unravel all the functions of the HO system.

  20. Expression, stabilization and purification of membrane proteins via diverse protein synthesis systems and detergents involving cell-free associated with self-assembly peptide surfactants. (United States)

    Zheng, Xuan; Dong, Shuangshuang; Zheng, Jie; Li, Duanhua; Li, Feng; Luo, Zhongli


    G-protein coupled receptors (GPCRs) are involved in regulating most of physiological actions and metabolism in the bodies, which have become most frequently addressed therapeutic targets for various disorders and diseases. Purified GPCR-based drug discoveries have become routine that approaches to structural study, novel biophysical and biochemical function analyses. However, several bottlenecks that GPCR-directed drugs need to conquer the problems including overexpression, solubilization, and purification as well as stabilization. The breakthroughs are to obtain efficient protein yield and stabilize their functional conformation which are both urgently requiring of effective protein synthesis system methods and optimal surfactants. Cell-free protein synthesis system is superior to the high yields and post-translation modifications, and early signs of self-assembly peptide detergents also emerged to superiority in purification of membrane proteins. We herein focus several predominant protein synthesis systems and surfactants involving the novel peptide detergents, and uncover the advantages of cell-free protein synthesis system with self-assembling peptide detergents in purification of functional GPCRs. This review is useful to further study in membrane proteins as well as the new drug exploration.

  1. ApoHRP-based Assay to Measure Intracellular Regulatory Heme (United States)

    Atamna, Hani; Brahmbhatt, Marmik; Atamna, Wafa; Shanower, Gregory A.; Dhahbi, Joseph M.


    The majority of the heme-binding proteins possess a “heme-pocket” that stably binds with heme. Usually known as housekeeping heme-proteins, they participate in a variety of metabolic reactions (e.g., catalase). Heme also binds with lower affinity to the “Heme-Regulatory Motifs” (HRM) in specific regulatory proteins. This type of heme binding is known as exchangeable or regulatory heme (RH). Heme binding to HRM proteins regulates their function (e.g., Bach1). Although there are well-established methods for assaying total cellular heme (e.g., heme-proteins plus RH), currently there is no method available for measuring RH independently from the total heme (TH). The current study describes and validates a new method to measure intracellular RH. The method is based on the reconstitution of apo-horseradish peroxidase (apoHRP) with heme to form holoHRP. The resulting holoHRP activity is then measured with a colorimetric substrate. The results show that apoHRP specifically binds RH but not with heme from housekeeping heme-proteins. The RH assay detects intracellular RH. Furthermore, using conditions that create positive (hemin) or negative (N-methyl protoporphyrin IX) controls for heme in normal human fibroblasts (IMR90), the RH assay shows that RH is dynamic and independent from TH. We also demonstrated that short-term exposure to subcytotoxic concentrations of lead (Pb), mercury (Hg), or amyloid-β(Aβ) significantly alters intracellular RH with little effect on TH. In conclusion the RH assay is an effective assay to investigate intracellular RH concentration and demonstrates that RH represents ~6% of total heme in IMR90 cells. PMID:25525887

  2. Regulation of intracellular heme trafficking revealed by subcellular reporters. (United States)

    Yuan, Xiaojing; Rietzschel, Nicole; Kwon, Hanna; Walter Nuno, Ana Beatriz; Hanna, David A; Phillips, John D; Raven, Emma L; Reddi, Amit R; Hamza, Iqbal


    Heme is an essential prosthetic group in proteins that reside in virtually every subcellular compartment performing diverse biological functions. Irrespective of whether heme is synthesized in the mitochondria or imported from the environment, this hydrophobic and potentially toxic metalloporphyrin has to be trafficked across membrane barriers, a concept heretofore poorly understood. Here we show, using subcellular-targeted, genetically encoded hemoprotein peroxidase reporters, that both extracellular and endogenous heme contribute to cellular labile heme and that extracellular heme can be transported and used in toto by hemoproteins in all six subcellular compartments examined. The reporters are robust, show large signal-to-background ratio, and provide sufficient range to detect changes in intracellular labile heme. Restoration of reporter activity by heme is organelle-specific, with the Golgi and endoplasmic reticulum being important sites for both exogenous and endogenous heme trafficking. Expression of peroxidase reporters in Caenorhabditis elegans shows that environmental heme influences labile heme in a tissue-dependent manner; reporter activity in the intestine shows a linear increase compared with muscle or hypodermis, with the lowest heme threshold in neurons. Our results demonstrate that the trafficking pathways for exogenous and endogenous heme are distinct, with intrinsic preference for specific subcellular compartments. We anticipate our results will serve as a heuristic paradigm for more sophisticated studies on heme trafficking in cellular and whole-animal models.

  3. Peroxisome proliferator-activated receptor alpha controls hepatic heme biosynthesis through ALAS1. (United States)

    Degenhardt, Tatjana; Väisänen, Sami; Rakhshandehroo, Maryam; Kersten, Sander; Carlberg, Carsten


    Heme is an essential prosthetic group of proteins involved in oxygen transport, energy metabolism and nitric oxide production. ALAS1 (5-aminolevulinate synthase) is the rate-limiting enzyme in heme synthesis in the liver and is highly regulated to adapt to the metabolic demand of the hepatocyte. In the present study, we describe human hepatic ALAS1 as a new direct target for the nuclear receptor peroxisome proliferator-activated receptor alpha (PPARalpha). In primary human hepatocytes and in HepG2 cells, PPARalpha agonists induced an increase in ALAS1 mRNA levels, which was abolished by PPARalpha silencing. These effects are mediated by two functional PPAR binding sites at positions -9 and -2.3 kb relative to the ALAS1 transcription start site. PPARalpha ligand treatment also up-regulated the mRNA levels of the genes ALAD (5-aminolevulinate dehydratase), UROS (uroporphyrinogen III synthase), UROD (uroporphyrinogen decarboxylase), CPOX (coproporphyrinogen oxidase) and PPOX (protoporphyrinogen oxidase) encoding for enzymes controlling further steps in heme biosynthesis. In HepG2 cells treated with PPARalpha agonists and in mouse liver upon fasting, the association of PPARalpha, its partner retinoid X receptor, PPARgamma co-activator 1alpha and activated RNA polymerase II with the transcription start site region of all six genes was increased, leading to higher levels of the metabolite heme. In conclusion, these data strongly support a role of PPARalpha in the regulation of human ALAS1 and of five additional genes of the pathway, consequently leading to increased heme synthesis.

  4. In vitro assembly of catalase. (United States)

    Baureder, Michael; Barane, Elisabeth; Hederstedt, Lars


    Most aerobic organisms contain catalase, which functions to decompose hydrogen peroxide. Typical catalases are structurally complex homo-tetrameric enzymes with one heme prosthetic group buried in each subunit. It is not known how catalase in the cell is assembled from its constituents. The bacterium Enterococcus faecalis cannot synthesize heme but can acquire it from the environment to form a cytoplasmic catalase. We have in E. faecalis monitored production of the enzyme polypeptide (KatA) depending on the availability of heme and used our findings to devise a procedure for the purification of preparative amounts of in vivo-synthesized apocatalase. We show that fully active catalase can be obtained in vitro by incubating isolated apoprotein with hemin. We have characterized features of the assembly process and describe a temperature-trapped hemylated intermediate of the enzyme maturation process. Hemylation of apocatalase does not require auxiliary cell components, but rapid assembly of active enzyme seemingly is assisted in the cell. Our findings provide insight about catalase assembly and offer new experimental possibilities for detailed studies of this process.

  5. Role of heme-protein covalent bonds in mammalian peroxidases. Protection of the heme by a single engineered heme-protein link in horseradish peroxidase. (United States)

    Huang, Liusheng; Wojciechowski, Grzegorz; Ortiz de Montellano, Paul R


    Oxidation of SCN-, Br-, and Cl- (X-) by horseradish peroxidase (HRP) and other plant and fungal peroxidases results in the addition of HOX to the heme vinyl group. This reaction is not observed with lactoperoxidase (LPO), in which the heme is covalently bound to the protein via two ester bonds between carboxylic side chains and heme methyl groups. To test the hypothesis that the heme of LPO and other mammalian peroxidases is protected from vinyl group modification by the hemeprotein covalent bonds, we prepared the F41E mutant of HRP in which the heme is attached to the protein via a covalent bond between Glu41 and the heme 3-methyl. We also examined the E375D mutant of LPO in which only one of the two normal covalent heme links is retained. The prosthetic heme groups of F41E HRP and E375D LPO are essentially not modified by the HOBr produced by these enzymes. The double E375D/D225E mutant of LPO that can form no covalent bonds is inactive and could not be examined. These results unambiguously demonstrate that a single heme-protein link is sufficient to protect the heme from vinyl group modification even in a protein (HRP) that is normally highly susceptible to this reaction. The results directly establish that one function of the covalent heme-protein bonds in mammalian peroxidases is to protect their prosthetic group from their highly reactive metabolic products.

  6. Hydrogen bonding discotic liquid crystals: Synthesis, self-assembly, and molecular recognition (United States)

    Bushey, Mark Lawrence

    The triamides shown below form discotic liquid crystalline phases with intermolecular hydrogen bonding stabilizing the columnar structure, A and B. The mesomorphic orientations of the columns are dependent on the amide side chain. Three mesophasic orientations are described: columns aligned perpendicular to the surface, columns aligned parallel to the surface in a radial pattern, and columns aligned parallel to the surface in a parallel or aligned pattern. The aggregation of the tridodecyloxy-triamides show N-H shifting in the IR at elevated temperatures, an indication that hydrogen bonding is important in the association of liquid crystalline mesophases. Powder X-ray diffraction studies indicate packing of the columns into a hexagonal lattice.* Studies on triamides with chiral side chains result in molecules stacking into columns displaying a helical pitch. In concentrated solutions of dodecane, molecules with chiral side chains display behavior consistent with chiral nematic liquid crystals; a super helical packing of the chiral columns. These superhelical packed systems show temperature dependent selective reflection of visible light and fingerprint textures. Atomic force microscopy (AFM) confirms in sub-monolayer films, that molecules preferring an edge-on orientation form long columns on highly ordered pyrolytic graphite (HOPG), those that prefer a face-on orientation form large amorphous domains. Electrostatic force microscopy (EFM) images of the domains of molecules in the edge-on orientation provides no discernible polarity, imaging of the domains of molecules in the face-on orientation indicates a negative polar orientation. Scanning probe measurements (SPM) of the tridodecynyl-triamide have shown similar edge-on orientations of other tridodecyloxy-triamides. Powder X-ray diffraction of these liquid crystalline phases shows a hexagonal packing of the columnar assembly. Electro-optic switching studies indicate a piezoelectric switching mechanism, possibly

  7. Independent evolution of four heme peroxidase superfamilies. (United States)

    Zámocký, Marcel; Hofbauer, Stefan; Schaffner, Irene; Gasselhuber, Bernhard; Nicolussi, Andrea; Soudi, Monika; Pirker, Katharina F; Furtmüller, Paul G; Obinger, Christian


    Four heme peroxidase superfamilies (peroxidase-catalase, peroxidase-cyclooxygenase, peroxidase-chlorite dismutase and peroxidase-peroxygenase superfamily) arose independently during evolution, which differ in overall fold, active site architecture and enzymatic activities. The redox cofactor is heme b or posttranslationally modified heme that is ligated by either histidine or cysteine. Heme peroxidases are found in all kingdoms of life and typically catalyze the one- and two-electron oxidation of a myriad of organic and inorganic substrates. In addition to this peroxidatic activity distinct (sub)families show pronounced catalase, cyclooxygenase, chlorite dismutase or peroxygenase activities. Here we describe the phylogeny of these four superfamilies and present the most important sequence signatures and active site architectures. The classification of families is described as well as important turning points in evolution. We show that at least three heme peroxidase superfamilies have ancient prokaryotic roots with several alternative ways of divergent evolution. In later evolutionary steps, they almost always produced highly evolved and specialized clades of peroxidases in eukaryotic kingdoms with a significant portion of such genes involved in coding various fusion proteins with novel physiological functions.

  8. Exploring genome-wide - dietary heme iron intake interactions and the risk of type 2 diabetes

    Directory of Open Access Journals (Sweden)

    Louis Robert Pasquale


    Full Text Available Aims/hypothesis: Genome-wide association studies have identified over 50 new genetic loci for type 2 diabetes (T2D. Several studies conclude that higher dietary heme iron intake increases the risk of T2D. Therefore we assessed whether the relation between genetic loci and type 2 diabetes is modified by dietary heme iron intake. Methods: We used Affymetrix Genome-Wide Human 6.0 array data (681,770 single nucleotide polymorphisms (SNPs and dietary information collected in the Health Professionals Follow-up Study (n=725 cases; n=1,273 controls and the Nurses’ Health Study (n=1,081 cases; n=1,692 controls. We assessed whether genome-wide SNPs or iron metabolism SNPs interacted with dietary heme iron intake in relation to T2D, testing for associations in each cohort separately and then meta-analyzing to pool the results. Finally, we created 1,000 synthetic pathways matched to an iron metabolism pathway on number of genes, and number of SNPs in each gene. We compared the iron metabolic pathway SNPs with these synthetic SNP assemblies in their relation to T2D to assess if the pathway as a whole interacts with dietary heme iron intake.Results: Using a genomic approach, we found no significant gene-environment interactions with dietary heme iron intake in relation to T2D (top SNP in pooled analysis: intergenic rs10980508; p=1.03E-06 > Bonferroni corrected p=7.33E-08. Furthermore, no SNP in the iron metabolic pathway significantly interacted with dietary heme iron intake (top SNP in pooled analysis: rs1805313; p=1.14E-03 > Bonferroni corrected p=2.10E-04. Finally, neither the main genetic effects (pooled empirical p by SNP=0.41, nor gene – dietary heme-iron interactions (pooled empirical p value for the interactions=0.72 were significant for the iron metabolic pathway as a whole. Conclusions: We found no significant interactions between dietary heme iron intake and common SNPs in relation to T2D.

  9. One-pot system for synthesis, assembly, and display of functional single-span membrane proteins on oil-water interfaces. (United States)

    Yunker, Peter J; Asahara, Haruichi; Hung, Kuo-Chan; Landry, Corey; Arriaga, Laura R; Akartuna, Ilke; Heyman, John; Chong, Shaorong; Weitz, David A


    Single-span membrane proteins (ssMPs) represent approximately one-half of all membrane proteins and play important roles in cellular communications. However, like all membrane proteins, ssMPs are prone to misfolding and aggregation because of the hydrophobicity of transmembrane helices, making them difficult to study using common aqueous solution-based approaches. Detergents and membrane mimetics can solubilize membrane proteins but do not always result in proper folding and functionality. Here, we use cell-free protein synthesis in the presence of oil drops to create a one-pot system for the synthesis, assembly, and display of functional ssMPs. Our studies suggest that oil drops prevent aggregation of some in vitro-synthesized ssMPs by allowing these ssMPs to localize on oil surfaces. We speculate that oil drops may provide a hydrophobic interior for cotranslational insertion of the transmembrane helices and a fluidic surface for proper assembly and display of the ectodomains. These functionalized oil drop surfaces could mimic cell surfaces and allow ssMPs to interact with cell surface receptors under an environment closest to cell-cell communication. Using this approach, we showed that apoptosis-inducing human transmembrane proteins, FasL and TRAIL, synthesized and displayed on oil drops induce apoptosis of cultured tumor cells. In addition, we take advantage of hydrophobic interactions of transmembrane helices to manipulate the assembly of ssMPs and create artificial clusters on oil drop surfaces. Thus, by coupling protein synthesis with self-assembly at the water-oil interface, we create a platform that can use recombinant ssMPs to communicate with cells.

  10. Heme Binding by Corynebacterium diphtheriae HmuT: Function and Heme Environment. (United States)

    Draganova, Elizabeth B; Akbas, Neval; Adrian, Seth A; Lukat-Rodgers, Gudrun S; Collins, Daniel P; Dawson, John H; Allen, Courtni E; Schmitt, Michael P; Rodgers, Kenton R; Dixon, Dabney W


    The heme uptake pathway (hmu) of Corynebacterium diphtheriae utilizes multiple proteins to bind and transport heme into the cell. One of these proteins, HmuT, delivers heme to the ABC transporter HmuUV. In this study, the axial ligation of the heme in ferric HmuT is probed by examination of wild-type (WT) HmuT and a series of conserved heme pocket residue mutants, H136A, Y235A, and M292A. Characterization by UV-visible, resonance Raman, and magnetic circular dichroism spectroscopies indicates that H136 and Y235 are the axial ligands in ferric HmuT. Consistent with this assignment of axial ligands, ferric WT and H136A HmuT are difficult to reduce while Y235A is reduced readily in the presence of dithionite. The FeCO Raman shifts in WT, H136A, and Y235A HmuT-CO complexes provide further evidence of the axial ligand assignments. Additionally, these frequencies provide insight into the nonbonding environment of the heme pocket. Ferrous Y235A and the Y235A-CO complex reveal that the imidazole of H136 exists in two forms, one neutral and one with imidazolate character, consistent with a hydrogen bond acceptor on the H136 side of the heme. The ferric fluoride complex of Y235A reveals the presence of at least one hydrogen bond donor on the Y235 side of the heme. Hemoglobin utilization assays showed that the axial Y235 ligand is required for heme uptake in HmuT.

  11. Kidney injury and heme oxygenase-1

    Directory of Open Access Journals (Sweden)

    Hai-xing MAI


    Full Text Available     Heme oxygenase-1 (HO-1 is one of the main pathways to degrade heme in mammals, and the main degradation products are free iron (Fe2+, carbon monoxide (CO, and bilirubin. Heme plays an important role in promoting cell survival, circulation of intracellular substrates, and immune regulation. Previous studies suggest that HO-1 pathway is an important internal factor in determining the susceptibility and severity of acute kidney injury (AKI. The induction of HO-1 expression can attenuate the severity of renal ischemia-reperfusion injury (IRI, and the inhibition of HO-1 expression will aggravate IRI. The present article summarizes the latest advances in research abroad and at home on protective mechanism by which HO-1 prevents AKI to further deepen our understanding of the role of HO-1 in the treatment of AKI.   

  12. Iminoguanidines as Allosteric Inhibitors of the Iron-Regulated Heme Oxygenase (HemO) of Pseudomonas aeruginosa



    New therapeutic targets are required to combat multidrug resistant infections, such as the iron-regulated heme oxygenase (HemO) of Pseudomonas aeruginosa, due to links between iron and virulence and dependence on heme as an iron source during infection. Herein we report the synthesis and activity of a series of iminoguanidine-based inhibitors of HemO. Compound 23 showed a binding affinity of 5.7 µM and an MIC50 of 52.3 µg/mL against P. aeruginosa PAO1. An in cellulo activity assay was develop...

  13. Red meat and colon cancer : how dietary heme initiates hyperproliferation

    NARCIS (Netherlands)

    IJssennagger, N.


    Colorectal cancer is a leading cause of cancer deaths in Western countries. The risk to develop colorectal cancer is associated with the intake of red meat. Red meat contains the porphyrin pigment heme. Heme is an irritant for the colonic wall and it is previously shown that the addition of heme to

  14. Molecular Simulations of Porphyrins and Heme Proteins

    Energy Technology Data Exchange (ETDEWEB)



    An overview of the use of classical mechanical molecular simulations of porphyrins, hydroporphyrins, and heme proteins is given. The topics cover molecular mechanics calculations of structures and conformer energies of porphyrins, energies of barriers for interconversion between stable conformers, molecular dynamics of porphyrins and heme proteins, and normal-coordinate structural analysis of experimental and calculated porphyrin structures. Molecular mechanics and dynamics are currently a fertile area of research on porphyrins. In the future, other computational methods such as Monte Carlo simulations, which have yet to be applied to porphyrins, will come into use and open new avenues of research into molecular simulations of porphyrins.

  15. Heme oxygenase-1 system and gastrointestinal tumors

    Institute of Scientific and Technical Information of China (English)

    Marie; CM; Lin; Hsiangfu; Kung


    Heme oxygenase-1(HO-1) system catabolizes heme into three products:carbon monoxide,biliverdin/bilirubin and free iron.It is involved in many physiological and pathophysiological processes.A great deal of data has demonstrated the roles of HO-1 in the formation,growth and metastasis of tumors.The interest in this system by investigators involved in gastrointestinal tumors is fairly recent,and few papers on HO-1 have touched upon this subject.This review focuses on the current understanding of the physiologic...

  16. Analysis of Heme oxygenase isomers in rat

    Institute of Scientific and Technical Information of China (English)

    Yun-ZhuLi; Wen-JunCui; Xue-HongZhang; Qing-XiangShen; JianWang; She


    AIM:To purify and identify heme oxygenase(HO) isomers which exist in rat liver,spleen and brain treated with hematin and phenylhydrazine and in untrated rat liver and to investigate the characteristics of HO isomers,to isolate and confirm the rat HO-1 cDNA that actually encodes HO-1 by expressing cDNA in monkey Kidney cells(COS-1 cells),to prepare the rat heme oxygenase-1(HO-1)mutant and to detect inhibition of HO-1 mutated enzyme.

  17. Synthesis and studies of polypeptide materials: Self-assembled block copolypeptide amphiphiles, DNA-condensing block copolypeptides and membrane-interactive random copolypeptides (United States)

    Wyrsta, Michael Dmytro

    A new class of transition metal initiators for the controlled polymerization of alpha-aminoacid-N-carboxyanhydrides (alpha-NCAs), has been developed by Deming et al. This discovery has allowed for the synthesis of well-defined "protein-like" polymers. Using this chemistry we have made distinct block/random copolypeptides for biomedical applications. Drug delivery, gene delivery, and antimicrobial polymers were the focus of our research efforts. The motivation for the synthesis and study of synthetic polypeptide based materials comes from proteins. Natural proteins are able to adopt a staggeringly large amount of uniquely well-defined folded structures. These structures account for the diversity in properties of proteins. As catalysts (enzymes) natural proteins perform some of the most difficult chemistry with ease and precision at ambient pressures and temperatures. They also exhibit incredible structural properties that directly result from formation of complex hierarchical assemblies. Self-assembling block copolymers were synthesized with various compositions and architectures. In general, di- and tri-block amphiphiles were studied for their self-assembling properties. Both spherical and tubular vesicles were found to assemble from di- and tri-block amphiphiles, respectively. In addition to self-assembly, pH responsiveness was engineered into these amphiphiles by the incorporation of basic residues (lysine) into the hydrophobic block. Another form of self-assembly studied was the condensation of DNA using cationic block copolymers. It was found that cationic block copolymers could condense DNA into compact, ordered, water-soluble aggregates on the nanoscale. These aggregates sufficiently protected DNA from nucleases and yet were susceptible to proteases. These studies form the basis of a gene delivery platform. The ease with which NCAs are polymerized renders them completely amenable to parallel synthetic methods. We have employed this technique to discover new

  18. Inhibition of Heme Peroxidases by Melamine

    Directory of Open Access Journals (Sweden)

    Pattaraporn Vanachayangkul


    Full Text Available In 2008 melamine-contaminated infant formula and dairy products in China led to over 50,000 hospitalizations of children due to renal injuries. In North America during 2007 and in Asia during 2004, melamine-contaminated pet food products resulted in numerous pet deaths due to renal failure. Animal studies have confirmed the potent renal toxicity of melamine combined with cyanuric acid. We showed previously that the solubility of melamine cyanurate is low at physiologic pH and ionic strength, provoking us to speculate how toxic levels of these compounds could be transported through the circulation without crystallizing until passing into the renal filtrate. We hypothesized that melamine might be sequestered by heme proteins, which could interfere with heme enzyme activity. Four heme peroxidase enzymes were selected for study: horseradish peroxidase (HRP, lactoperoxidase (LPO, and cyclooxygenase-1 and -2 (COX-1 and -2. Melamine exhibited noncompetitive inhibition of HRP (9.5±0.7mM, and LPO showed a mixed model of inhibition (14.5±4.7mM. The inhibition of HRP and LPO was confirmed using a chemiluminescent peroxidase assay. Melamine also exhibited COX-1 inhibition, but inhibition of COX-2 was not detected. Thus, our results demonstrate that melamine inhibits the activity of three heme peroxidases.

  19. Co-Assembled Supported Catalysts: Synthesis of Nano-Structured Supported Catalysts with Hierarchic Pores through Combined Flow and Radiation Induced Co-Assembled Nano-Reactors


    Galip Akay


    A novel generic method of silica supported catalyst system generation from a fluid state is presented. The technique is based on the combined flow and radiation (such as microwave, thermal or UV) induced co-assembly of the support and catalyst precursors forming nano-reactors, followed by catalyst precursor decomposition. The transformation from the precursor to supported catalyst oxide state can be controlled from a few seconds to several minutes. The resulting nano-structured micro-porous s...

  20. Activation of lactoperoxidase by heme-linked protonation and heme-independent iodide binding. (United States)

    Toyama, Akira; Tominaga, Aya; Inoue, Tatsuo; Takeuchi, Hideo


    Lactoperoxidase (LPO), a mammalian secretory heme peroxidase, catalyzes the oxidation of thiocyanate by hydrogen peroxide to produce hypothiocyanate, an antibacterial agent. Although LPO is known to be activated at acidic pH and in the presence of iodide, the structural basis of the activation is not well understood. We have examined the effects of pH and iodide concentration on the catalytic activity and the structure of LPO. Electrochemical and colorimetric assays have shown that the catalytic activity is maximized at pH 4.5. The heme Soret absorption band exhibits a small red-shift at pH 5.0 upon acidification, which is ascribable to a structural transition from a neutral to an acidic form. Resonance Raman spectra suggest that the heme porphyrin core is slightly contracted and the Fe-His bond is strengthened in the acidic form compared to the neutral form. The structural change of LPO upon activation at acidic pH is similar to that observed for myeloperoxidase, another mammalian heme peroxidase, upon activation at neutral pH. Binding of iodide enhances the catalytic activity of LPO without affecting either the optimum pH of activity or the heme structure, implying that the iodide binding occurs at a protein site away from the heme-linked protonation site.

  1. Lessons from bloodless worms: heme homeostasis in C. elegans. (United States)

    Sinclair, Jason; Hamza, Iqbal


    Heme is an essential cofactor for proteins involved in diverse biological processes such as oxygen transport, electron transport, and microRNA processing. Free heme is hydrophobic and cytotoxic, implying that specific trafficking pathways must exist for the delivery of heme to target hemoproteins which reside in various subcellular locales. Although heme biosynthesis and catabolism have been well characterized, the pathways for trafficking heme within and between cells remain poorly understood. Caenorhabditis elegans serves as a unique animal model for uncovering these pathways because, unlike vertebrates, the worm lacks enzymes to synthesize heme and therefore is crucially dependent on dietary heme for sustenance. Using C. elegans as a genetic animal model, several novel heme trafficking molecules have been identified. Importantly, these proteins have corresponding homologs in vertebrates underscoring the power of using C. elegans, a bloodless worm, in elucidating pathways in heme homeostasis and hematology in humans. Since iron deficiency and anemia are often exacerbated by parasites such as helminths and protozoa which also rely on host heme for survival, C. elegans will be an ideal model to identify anti-parasitic drugs that target heme transport pathways unique to the parasite.

  2. Assembling Synthesis of ZnSe Orthohexagonal Slices through Emulsion Liquid Membrane System of Gas-liquid Transport

    Institute of Scientific and Technical Information of China (English)

    Lu LIU; Qing Sheng WU; Ya Ping DING; Hua Jie LIU


    Orthohexagonal slices assembled by ZnSe quantum dots were synthesized through emulsion liquid membrane system. These orthohexagonal slices were 1.5-3.5 μm in side length and were self-assembled by ZnSe quantum dots of 2-3 nm. It was proposed the surfactant molecules on ZnSe quantum dots played a key role in the self-assembly process.

  3. A novel water-soluble anionic conjugated copolymer containing poly(p-phenylene vinylene) segments: Copolymer synthesis and multilayer construction by assembling poly(diallyl dimethyl ammonium chloride)

    Institute of Scientific and Technical Information of China (English)

    ZHANG Bao; LIN Hang; TANG Jun; LIU JunSong


    This paper reports the synthesis of a water soluble conjugated polymer poly(p-phenylene vinylene-co-sodium methacrylate) (ws-P(PV-co-SMA)) and the multilayer of the derived copolymer assembling poly(diallyl dimethyl ammanium chloride) (PDDA). The self-assembling process of the mulUlayer was monitored by UV-vis absorption spectroscopy, and the data indicated a linear increase in film thick-ness with a number of ws-P(PV-co-SMA)/PDDA bilayers. The alternative deposition of ws-P (PV-co-SMA) and PDDA allowed the insertion of a non-conjugated layer between the conjugated layers, thus the migration of the photogenerated polarons was effectively confined in the isolated ws-P (PV-co-SMA) chains. Consequently, the photoluminescence quantum yield reached 0.68, 30 times higher than that of pure poly(p-phenylen vinylene). The distinct electronic interactions between conjugated segments were confirmed by comparative analyses of the excitation spectra and time-resolved photolumines-cence spectra of ws-P(PV-co-SMA) solid film and the assembled multilayers. The confinement effect of the PDDA layer on the photogenerated carriers was verified by the surface photovoltage spectroscopic measurement on both ws-P(PV-co-SMA) solid film and self-assembled multilayers.

  4. Toward a modular multi-material nanoparticle synthesis and assembly strategy via bionanocombinatorics: bifunctional peptides for linking Au and Ag nanomaterials

    Energy Technology Data Exchange (ETDEWEB)

    Briggs, Beverly D.; Palafox-Hernandez, J. Pablo; Li, Yue; Lim, Chang-Keun; Woehl, Taylor J.; Bedford, Nicholas M.; Seifert, Soenke; Swihart, Mark T.; Prasad, Paras N.; Walsh, Tiffany R.; Knecht, Marc R.


    Materials-binding peptides represent a unique avenue towards controlling the shape and size of nanoparticles (NPs) grown under aqueous conditions. Here, employing a bionanocombinatorics approach, two such materials-binding peptides were linked at either end of a photoswitchable spacer, forming a multi-domain materials-binding molecule to control the in situ synthesis and organization of Ag and Au NPs under ambient conditions. These multi-domain molecules retained the peptides’ ability to nucleate, grow, and stabilize Ag and Au NPs in aqueous media. Disordered co-assemblies of the two nanomaterials were observed by TEM imaging of dried samples after sequential growth of the two metals, and showed a clustering behavior that was not observed without both metals and the linker molecules. While TEM evidence indicated the formation of AuNP/AgNP assemblies upon drying, SAXS analysis indicated that no extended assemblies existed in solution, suggesting that sample drying plays an important role in facilitating NP clustering. Molecular simulations and experimental data revealed tunable materials-binding based upon the isomerization state of the photoswitchable unit and metal employed. This work is a first step in generating externally actuated biomolecules with specific material-binding properties that could be used as the building blocks to achieve multi-material switchable NP assemblies.

  5. Synthesis of new tren-based tris-macrocycles. Anion cluster assembling inside the cavity generated by a bowl-shaped receptor. (United States)

    Bazzicalupi, Carla; Bencini, Andrea; Berni, Emanuela; Bianchi, Antonio; Ciattini, Samuele; Giorgi, Claudia; Maoggi, Sauro; Paoletti, Piero; Valtancoli, Barbara


    The synthesis of three new tris-macrocycles, containing three [12]aneN(4) (L1), [12]aneN(3)O (L2), or [14]aneN(4) (L3) moieties appended to a tren unit, is reported. The crystal structure of the [(Na(ClO(4))(6)) subset L1(2)H(13)]Na(6)Cl(2)(ClO(4))(12) compound shows the anionic cluster [Na(ClO(4))(6)](5)(-) assembled inside the cavity defined by two bowl-shaped polyammonium receptors, held by multiple charge-charge and hydrogen bond interactions.

  6. Activation of heme oxygenase and consequent carbon monoxide formation inhibits the release of arginine vasopressin from rat hypothalamic explants. Molecular linkage between heme catabolism and neuroendocrine function. (United States)

    Mancuso, C; Kostoglou-Athanassiou, I; Forsling, M L; Grossman, A B; Preziosi, P; Navarra, P; Minotti, G


    Heme oxygenase (HO)-catalyzed degradation of cellular heme moieties generates biliverdin and equimolar amounts of carbon monoxide (CO), which has been implicated as a possible modulator of neural function. Technical difficulties preclude direct measurements of CO within intact nervous tissues; hence, alternative procedures are needed to monitor the formation and possible biologic functions of this gas. In the present study rat hypothalamic explants were found to generate 114 +/- 5 or 127 +/- 11 pmol biliverdin/hypothalamus/1 h (n = 3) upon incubation with 1 or 10 microM hemin, respectively. Ten micromolar zinc-protoporphyrin IX (Zn-PP-IX), a known inhibitor of HO, significantly decreased the degradation of 10 microM hemin from 127 +/- 11 to 26 +/- 11 pmol biliverdin/hypothalamus/1 h (n = 3; P tin-mesoporphyrin IX, which is even more selective in inhibiting HO; it was also attenuated in the presence of the gaseous scavenger ferrous hemoglobin. Furthermore, the inhibition of AVP release could be reproduced by omitting hemin and by incubating hypothalami under CO, whereas treatment with biliverdin had no effect. This suggested that the release of AVP was suppressed by HO degradation of hemin, yielding CO as a modulator of hypothalamic function. These observations may be relevant to diseases characterized by inappropriate secretion of AVP and enzymatic disturbances affecting the synthesis of heme and the formation of CO through the HO pathway (e.g., acute intermittent porphyria or lead intoxication).

  7. Induction of heme oxygenase 1 by nitrosative stress. A role for nitroxyl anion. (United States)

    Naughton, Patrick; Foresti, Roberta; Bains, Sandip K; Hoque, Martha; Green, Colin J; Motterlini, Roberto


    Nitric oxide and S-nitrosothiols modulate a variety of important physiological activities. In vascular cells, agents that release NO and donate nitrosonium cation (NO(+)), such as S-nitrosoglutathione, are potent inducers of the antioxidant protein heme oxygenase 1 (HO-1) (Foresti, R., Clark, J. E., Green, C. J., and Motterlini, R. (1997) J. Biol. Chem. 272, 18411-18417; Motterlini, R., Foresti, R., Bassi, R., Calabrese, V., Clark, J. E., and Green, C. J. (2000) J. Biol. Chem. 275, 13613-13620). Here, we report that Angeli's salt (AS) (0.25-2 mm), a compound that releases nitroxyl anion (NO(-)) at physiological pH, induces HO-1 mRNA and protein expression in a concentration- and time-dependent manner, resulting in increased heme oxygenase activity in rat H9c2 cells. A time course analysis revealed that NO(-)-mediated HO-1 expression is transient and gradually disappears within 24 h, in accordance with the short half-life of AS at 37 degrees C (t(12) = 2.3 min). Interestingly, multiple additions of AS at lower concentrations (50 or 100 microm) over a period of time still promoted a significant increase in heme oxygenase activity. Experiments performed using a NO scavenger and the NO electrode confirmed that NO(-), not NO, is the species involved in HO-1 induction by AS; however, the effect on heme oxygenase activity can be amplified by accelerating the rate of NO(-) oxidation. N-Acetylcysteine almost completely abolished AS-mediated induction of HO-1, whereas a glutathione synthesis inhibitor (buthionine sulfoximine) significantly decreased heme oxygenase activation by AS, indicating that sulfydryl groups are crucial targets in the regulation of HO-1 expression by NO(-). We conclude that NO(-), in analogy with other reactive nitrogen species, is a potent inducer of heme oxygenase activity and HO-1 protein expression. These findings indicate that heme oxygenase can act both as a sensor to and target of redox-based mechanisms involving NO and extend our knowledge on

  8. Identification of essential histidine residues involved in heme binding and Hemozoin formation in heme detoxification protein from Plasmodium falciparum. (United States)

    Nakatani, Keisuke; Ishikawa, Haruto; Aono, Shigetoshi; Mizutani, Yasuhisa


    Malaria parasites digest hemoglobin within a food vacuole to supply amino acids, releasing the toxic product heme. During the detoxification, toxic free heme is converted into an insoluble crystalline form called hemozoin (Hz). Heme detoxification protein (HDP) in Plasmodium falciparum is one of the most potent of the hemozoin-producing enzymes. However, the reaction mechanisms of HDP are poorly understood. We identified the active site residues in HDP using a combination of Hz formation assay and spectroscopic characterization of mutant proteins. Replacement of the critical histidine residues His122, His172, His175, and His197 resulted in a reduction in the Hz formation activity to approximately 50% of the wild-type protein. Spectroscopic characterization of histidine-substituted mutants revealed that His122 binds heme and that His172 and His175 form a part of another heme-binding site. Our results show that the histidine residues could be present in the individual active sites and could be ligated to each heme. The interaction between heme and the histidine residues would serve as a molecular tether, allowing the proper positioning of two hemes to enable heme dimer formation. The heme dimer would act as a seed for the crystal growth of Hz in P. falciparum.

  9. Convenient synthesis and application of versatile nucleic acid lipid membrane anchors in the assembly and fusion of liposomes

    DEFF Research Database (Denmark)

    Ries, Oliver; Löffler, Philipp M. G.; Vogel, Stefan


    Hydrophobic moieties like lipid membrane anchors are highly demanded modifications for nucleic acid oligomers. Membrane-anchor modified oligonucleotides are applicable in biomedicine leading to new delivery strategies as well as in biophysical investigations towards assembly and fusion of liposom...

  10. Synthesis, morphological control, dispersion stabilization and in situ self-assembly of noble metal nanostructures using multidentate resorcinarene surfactants (United States)

    Han, Sangbum

    In this dissertation, a detailed investigation on the influence of various macrocyclic resorcinarene surfactants in determining the morphology, stabilization and self-assembly of mono- and bi- metallic nanoparticles was undertaken. (Abstract shortened by ProQuest.).

  11. Synthesis and assembly with mesoporous silica of platinum (II) porphyrin complexes bearing carbazyl groups: Luminescent and oxygen sensing properties

    Institute of Scientific and Technical Information of China (English)

    HUO Cheng; ZHANG Huidong; GUO Jianhua; ZHANG Hongyu; ZHANG Ping; WANG Yue


    A series of platinum meso-tetrakis [3-methoxy-4-(N-carbazyl)n-alkyloxyphenyl]porphyrin (Pt-4Cn-TPP, n = 4, 6 and 8) are synthesized. Pt-4C4-TPP, Pt-4C6-TPP and Pt-4C8-TPP exhibit similar luminescent properties in solution and solid state. Three protonated platinum (II) porphyrins are assembled with mesoporous silica MCM-48, respectively, resulting in assembly materials Pt-4Cn-TPP4+/ MCM-48 (n = 4, 6 and 8). The luminescent intensity of Pt-4Cn-TPP4+/MCM-48 can be extremely quenched by molecular oxygen with high sensitivity (I0/I100>9). The Stern-Volmer plots of these assembly materials display considerable linearity within a wide range of oxygen concentration (0 to 100%). The response time is all ≤ 1 s and recovery time ≤ 22 s for these assembly materials.

  12. Determining the Subcellular Location of Synthesis and Assembly of the Cell Wall Polysaccharide (1,3; 1,4)-β-d-Glucan in Grasses[OPEN (United States)

    Wilson, Sarah M.; Ho, Yin Ying; Lampugnani, Edwin R.; Van de Meene, Allison M.L.; Bain, Melissa P.; Bacic, Antony; Doblin, Monika S.


    The current dogma for cell wall polysaccharide biosynthesis is that cellulose (and callose) is synthesized at the plasma membrane (PM), whereas matrix phase polysaccharides are assembled in the Golgi apparatus. We provide evidence that (1,3;1,4)-β-d-glucan (mixed-linkage glucan [MLG]) does not conform to this paradigm. We show in various grass (Poaceae) species that MLG-specific antibody labeling is present in the wall but absent over Golgi, suggesting it is assembled at the PM. Antibodies to the MLG synthases, cellulose synthase-like F6 (CSLF6) and CSLH1, located CSLF6 to the endoplasmic reticulum, Golgi, secretory vesicles, and the PM and CSLH1 to the same locations apart from the PM. This pattern was recreated upon expression of VENUS-tagged barley (Hordeum vulgare) CSLF6 and CSLH1 in Nicotiana benthamiana leaves and, consistent with our biochemical analyses of native grass tissues, shown to be catalytically active with CSLF6 and CSLH1 in PM-enriched and PM-depleted membrane fractions, respectively. These data support a PM location for the synthesis of MLG by CSLF6, the predominant enzymatically active isoform. A model is proposed to guide future experimental approaches to dissect the molecular mechanism(s) of MLG assembly. PMID:25770111

  13. Role of Heme and Heme-Proteins in Trypanosomatid Essential Metabolic Pathways

    Directory of Open Access Journals (Sweden)

    Karina E. J. Tripodi


    Full Text Available Around the world, trypanosomatids are known for being etiological agents of several highly disabling and often fatal diseases like Chagas disease (Trypanosoma cruzi, leishmaniasis (Leishmania spp., and African trypanosomiasis (Trypanosoma brucei. Throughout their life cycle, they must cope with diverse environmental conditions, and the mechanisms involved in these processes are crucial for their survival. In this review, we describe the role of heme in several essential metabolic pathways of these protozoans. Notwithstanding trypanosomatids lack of the complete heme biosynthetic pathway, we focus our discussion in the metabolic role played for important heme-proteins, like cytochromes. Although several genes for different types of cytochromes, involved in mitochondrial respiration, polyunsaturated fatty acid metabolism, and sterol biosynthesis, are annotated at the Tritryp Genome Project, the encoded proteins have not yet been deeply studied. We pointed our attention into relevant aspects of these protein functions that are amenable to be considered for rational design of trypanocidal agents.

  14. Hydrothermal Synthesis and Crystal Structure of Manganese(Ⅱ) Coordination Polymer Assembled by 4-Sulfophthalate and 4,4'-Bipyridine

    Institute of Scientific and Technical Information of China (English)


    @@ Introduction The rational design and synthesis of metal-directed supramolecular framework compounds have received much attention in coordination chemistry because of their potential applications in catalysis, molecular selection, nonlinear optics, ion exchange, and microelectronics[1-4].

  15. Isocyanides Inhibit Human Heme Oxygenases at the Verdoheme Stage†


    Evans, John P.; Kandel, Sylvie; Ortiz de Montellano, Paul R.


    Heme oxygenases (HO) catalyze the oxidative cleavage of heme to generate biliverdin, CO, and free iron. In humans, heme oxygenase-1 (hHO-1) is overexpressed in tumor tissues, where it helps to protect cancer cells from anticancer agents, while HOs in fungal pathogens, such as Candida albicans, function as the primary means of iron acquisition. Thus, HO can be considered a potential therapeutic target for certain diseases. In this study, we have examined the equilibrium binding of three isocya...

  16. Holo- And Apo- Structures of Bacterial Periplasmic Heme Binding Proteins

    Energy Technology Data Exchange (ETDEWEB)

    Ho, W.W.; Li, H.; Eakanunkul, S.; Tong, Y.; Wilks, A.; Guo, M.; Poulos, T.L.


    An essential component of heme transport in Gram-negative bacterial pathogens is the periplasmic protein that shuttles heme between outer and inner membranes. We have solved the first crystal structures of two such proteins, ShuT from Shigella dysenteriae and PhuT from Pseudomonas aeruginosa. Both share a common architecture typical of Class III periplasmic binding proteins. The heme binds in a narrow cleft between the N- and C-terminal binding domains and is coordinated by a Tyr residue. A comparison of the heme-free (apo) and -bound (holo) structures indicates little change in structure other than minor alterations in the heme pocket and movement of the Tyr heme ligand from an 'in' position where it can coordinate the heme iron to an 'out' orientation where it points away from the heme pocket. The detailed architecture of the heme pocket is quite different in ShuT and PhuT. Although Arg{sup 228} in PhuT H-bonds with a heme propionate, in ShuT a peptide loop partially takes up the space occupied by Arg{sup 228}, and there is no Lys or Arg H-bonding with the heme propionates. A comparison of PhuT/ShuT with the vitamin B{sub 12}-binding protein BtuF and the hydroxamic-type siderophore-binding protein FhuD, the only two other structurally characterized Class III periplasmic binding proteins, demonstrates that PhuT/ShuT more closely resembles BtuF, which reflects the closer similarity in ligands, heme and B{sub 12}, compared with ligands for FhuD, a peptide siderophore.

  17. Mechanisms of peroxynitrite interactions with heme proteins. (United States)

    Su, Jia; Groves, John T


    Oxygenated heme proteins are known to react rapidly with nitric oxide (NO) to produce peroxynitrite (PN) at the heme site. This process could lead either to attenuation of the effects of NO or to nitrosative protein damage. PN is a powerful nitrating and oxidizing agent that has been implicated in a variety of cell injuries. Accordingly, it is important to delineate the nature and variety of reaction mechanisms of PN interactions with heme proteins. In this Forum, we survey the range of reactions of PN with heme proteins, with particular attention to myoglobin and cytochrome c. While these two proteins are textbook paradigms for oxygen binding and electron transfer, respectively, both have recently been shown to have other important functions that involve NO and PN. We have recently described direct evidence that ferrylmyolgobin (ferrylMb) and nitrogen dioxide (NO(2)) are both produced during the reaction of PN and metmyolgobin (metMb) (Su, J.; Groves, J. T. J. Am. Chem. Soc. 2009, 131, 12979-12988). Kinetic evidence indicates that these products evolve from the initial formation of a caged radical intermediate [Fe(IV) horizontal lineO.NO(2)]. This caged pair reacts mainly via internal return with a rate constant k(r) to form metMb and nitrate in an oxygen-rebound scenario. Detectable amounts of ferrylMb are observed by stopped-flow spectrophotometry, appearing at a rate consistent with the rate, k(obs), of heme-mediated PN decomposition. Freely diffusing NO(2), which is liberated concomitantly from the radical pair (k(e)), preferentially nitrates myoglobin Tyr103 and added fluorescein. For cytochrome c, Raman spectroscopy has revealed that a substantial fraction of cytochrome c converts to a beta-sheet structure, at the expense of turns and helices at low pH (Balakrishnan, G.; Hu, Y.; Oyerinde, O. F.; Su, J.; Groves, J. T.; Spiro, T. G. J. Am. Chem. Soc., 2007, 129, 504-505). It is proposed that a short beta-sheet segment, comprising residues 37-39 and 58

  18. π-Extended Star-Shaped Polycyclic Aromatic Hydrocarbons based on Fused Truxenes: Synthesis, Self-Assembly, and Facilely Tunable Emission Properties. (United States)

    Cheng, Cheng; Jiang, Yi; Liu, Cheng-Fang; Zhang, Jian-Dong; Lai, Wen-Yong; Huang, Wei


    A new set of star-shaped polycyclic aromatic hydrocarbons (PAHs) based on naphthalene-fused truxenes, TrNaCn (n=1-4), were synthesized and characterized. The synthesis involved a microwave-assisted six-fold Suzuki coupling reaction, followed by oxidative cyclodehydrogenation. Multiple dehydrocyclization products could be effectively isolated in a single reaction, thus suggesting that the oxidative cyclodehydrogenation reaction involved a stepwise ring-closing process. The thermal, optical, and electrochemical properties and the self-assembly behavior of the resulting oxidized samples were investigated to understand the impact of the ring-fusing process on the properties of the star-shaped PAHs. Distinct bathochromic shift of the absorption maxima (λmax ) revealed that the molecular conjugation extended with the stepwise ring-closing reactions. The optical band-gap energy of these PAHs varied significantly on increasing the number of fused rings, thereby resulting in readily tunable emissive properties of the resultant star-shaped PAHs. Interestingly, the generation of rigid "arms" by using perylene analogues caused TrNaC2 and TrNaC3 to show significantly enhanced photoluminescence quantum yields (PLQYs) in solution (η=0.65 and 0.66, respectively) in comparison with those of TrNa and TrNaC1 (η=0.08 and 0.16, respectively). Owing to strong intermolecular interactions, the TrNa precursor was able to self-assemble into rod-like microcrystals, which could be facilely identified by the naked eye, whilst TrNaC1 self-assembled into nanosheets once the naphthalene rings had fused. This study offers a unique platform to gain further insight into-and a better understanding of-the photophysical and self-assembly properties of π-extended star-shaped PAHs.

  19. Co-Assembled Supported Catalysts: Synthesis of Nano-Structured Supported Catalysts with Hierarchic Pores through Combined Flow and Radiation Induced Co-Assembled Nano-Reactors

    Directory of Open Access Journals (Sweden)

    Galip Akay


    Full Text Available A novel generic method of silica supported catalyst system generation from a fluid state is presented. The technique is based on the combined flow and radiation (such as microwave, thermal or UV induced co-assembly of the support and catalyst precursors forming nano-reactors, followed by catalyst precursor decomposition. The transformation from the precursor to supported catalyst oxide state can be controlled from a few seconds to several minutes. The resulting nano-structured micro-porous silica supported catalyst system has a surface area approaching 300 m2/g and X-ray Diffraction (XRD-based catalyst size controlled in the range of 1–10 nm in which the catalyst structure appears as lamellar sheets sandwiched between the catalyst support. These catalyst characteristics are dependent primarily on the processing history as well as the catalyst (Fe, Co and Ni studied when the catalyst/support molar ratio is typically 0.1–2. In addition, Ca, Mn and Cu were used as co-catalysts with Fe and Co in the evaluation of the mechanism of catalyst generation. Based on extensive XRD, Scanning Electron Microscopy (SEM and Transmission Electron Microscopy (TEM studies, the micro- and nano-structure of the catalyst system were evaluated. It was found that the catalyst and silica support form extensive 0.6–2 nm thick lamellar sheets of 10–100 nm planar dimensions. In these lamellae, the alternate silica support and catalyst layer appear in the form of a bar-code structure. When these lamellae structures pack, they form the walls of a micro-porous catalyst system which typically has a density of 0.2 g/cm3. A tentative mechanism of catalyst nano-structure formation is provided based on the rheology and fluid mechanics of the catalyst/support precursor fluid as well as co-assembly nano-reactor formation during processing. In order to achieve these structures and characteristics, catalyst support must be in the form of silane coated silica nano

  20. High-definition self-assemblies driven by the hydrophobic effect: synthesis and properties of a supramolecular nanocapsule. (United States)

    Liu, Simin; Gibb, Bruce C


    High definition self-assemblies, those that possess order at the molecular level, are most commonly made from subunits possessing metals and metal coordination sites, or groups capable of partaking in hydrogen bonding. In other words, enthalpy is the driving force behind the free energy of assembly. The hydrophobic effect engenders the possibility of (nominally) relying not on enthalpy but entropy to drive assembly. Towards this idea, we describe how template molecules can trigger the dimerization of a cavitand in aqueous solution, and in doing so are encapsulated within the resulting capsule. Although not held together by (enthalpically) strong and directional non-covalent forces, these capsules possess considerable thermodynamic and kinetic stability. As a result, they display unusual and even unique properties. We discuss some of these, including the use of the capsule as a nanoscale reaction chamber and how they can bring about the separation of hydrocarbon gases.

  1. ex vivo DNA assembly

    Directory of Open Access Journals (Sweden)

    Adam B Fisher


    Full Text Available Even with decreasing DNA synthesis costs there remains a need for inexpensive, rapid and reliable methods for assembling synthetic DNA into larger constructs or combinatorial libraries. Advances in cloning techniques have resulted in powerful in vitro and in vivo assembly of DNA. However, monetary and time costs have limited these approaches. Here, we report an ex vivo DNA assembly method that uses cellular lysates derived from a commonly used laboratory strain of Escherichia coli for joining double-stranded DNA with short end homologies embedded within inexpensive primers. This method concurrently shortens the time and decreases costs associated with current DNA assembly methods.

  2. CYB5D2 requires heme-binding to regulate HeLa cell growth and confer survival from chemotherapeutic agents.

    Directory of Open Access Journals (Sweden)

    Anthony Bruce

    Full Text Available The cytochrome b5 domain containing 2 (CYB5D2; Neuferricin protein has been reported to bind heme, however, the critical residues responsible for heme-binding are undefined. Furthermore, the relationship between heme-binding and CYB5D2-mediated intracellular functions remains unknown. Previous studies examining heme-binding in two cytochrome b5 heme-binding domain-containing proteins, damage-associated protein 1 (Dap1; Saccharomyces cerevisiae and human progesterone receptor membrane component 1 (PGRMC1, have revealed that conserved tyrosine (Y 73, Y79, aspartic acid (D 86, and Y127 residues present in human CYB5D2 may be involved in heme-binding. CYB5D2 binds to type b heme, however, only the substitution of glycine (G at D86 (D86G within its cytochrome b5 heme-binding (cyt-b5 domain abolished its heme-binding ability. Both CYB5D2 and CYB5D2(D86G localize to the endoplasmic reticulum. Ectopic CYB5D2 expression inhibited cell proliferation and anchorage-independent colony growth of HeLa cells. Conversely, CYB5D2 knockdown and ectopic CYB5D2(D86G expression increased cell proliferation and colony growth. As PGRMC1 has been reported to regulate the expression and activities of cytochrome P450 proteins (CYPs, we examined the role of CYB5D2 in regulating the activities of CYPs involved in sterol synthesis (CYP51A1 and drug metabolism (CYP3A4. CYB5D2 co-localizes with cytochrome P450 reductase (CYPOR, while CYB5D2 knockdown reduced lanosterol demethylase (CYP51A1 levels and rendered HeLa cells sensitive to mevalonate. Additionally, knockdown of CYB5D2 reduced CYP3A4 activity. Lastly, CYB5D2 expression conferred HeLa cell survival from chemotherapeutic agents (paclitaxel, cisplatin and doxorubicin, with its ability to promote survival being dependent on its heme-binding ability. Taken together, this study provides evidence that heme-binding is critical for CYB5D2 in regulating HeLa cell growth and survival, with endogenous CYB5D2 being required to

  3. Synthesis, self-assembly and photoinduced surface-relief gratings of a polyacrylate-based Azo polyelectrolyte (United States)

    He, Yaning; Wang, Haopeng; Tuo, Xinlin; Deng, Wei; Wang, Xiaogong


    A polyacrylate-based azo polyelectrolyte was synthesized and characterized by the spectroscopic methods and thermal analysis. Layer-by-layer self-assembly of the azo polyelectrolyte through electrostatic adsorption was explored. By using a dipping solution of the anionic azo polyelectrolyte in anhydrous DMF, together with an aqueous solution of cationic poly(diallyldimethylammonium chloride) (PDAC), high quality multilayer films were obtained through the sequential deposition of the oppositely charged polyelectrolytes. With interfering illumination of Ar + laser beams (488 nm), significant surface-relief gratings formed on the self-assembled multiplayer films were observed.

  4. Heme Recognition By a Staphylococcus Aureus IsdE

    Energy Technology Data Exchange (ETDEWEB)

    Grigg, J.C.; Vermeiren, C.L.; Heinrichs, D.E.; Murphy, M.E.P.


    Staphylococcus aureus is a Gram-positive bacterial pathogen and a leading cause of hospital acquired infections. Because the free iron concentration in the human body is too low to support growth, S. aureus must acquire iron from host sources. Heme iron is the most prevalent iron reservoir in the human body and a predominant source of iron for S. aureus. The iron-regulated surface determinant (Isd) system removes heme from host heme proteins and transfers it to IsdE, the cognate substrate-binding lipoprotein of an ATP-binding cassette transporter, for import and subsequent degradation. Herein, we report the crystal structure of the soluble portion of the IsdE lipoprotein in complex with heme. The structure reveals a bi-lobed topology formed by an N- and C-terminal domain bridged by a single {alpha}-helix. The structure places IsdE as a member of the helical backbone metal receptor superfamily. A six-coordinate heme molecule is bound in the groove established at the domain interface, and the heme iron is coordinated in a novel fashion for heme transporters by Met{sup 78} and His{sup 229}. Both heme propionate groups are secured by H-bonds to IsdE main chain and side chain groups. Of these residues, His{sup 299} is essential for IsdE-mediated heme uptake by S. aureus when growth on heme as a sole iron source is measured. Multiple sequence alignments of homologues from several other Gram-positive bacteria, including the human pathogens pyogenes, Bacillus anthracis, and Listeria monocytogenes, suggest that these other systems function equivalently to S. aureus IsdE with respect to heme binding and transport.

  5. Colloidal Gold--Collagen Protein Core--Shell Nanoconjugate: One-Step Biomimetic Synthesis, Layer-by-Layer Assembled Film, and Controlled Cell Growth. (United States)

    Xing, Ruirui; Jiao, Tifeng; Yan, Linyin; Ma, Guanghui; Liu, Lei; Dai, Luru; Li, Junbai; Möhwald, Helmuth; Yan, Xuehai


    The biogenic synthesis of biomolecule-gold nanoconjugates is of key importance for a broad range of biomedical applications. In this work, a one-step, green, and condition-gentle strategy is presented to synthesize stable colloidal gold-collagen core-shell nanoconjugates in an aqueous solution at room temperature, without use of any reducing agents and stabilizing agents. It is discovered that electrostatic binding between gold ions and collagen proteins and concomitant in situ reduction by hydroxyproline residues are critically responsible for the formation of the core-shell nanoconjugates. The film formed by layer-by-layer assembly of such colloidal gold-collagen nanoconjugates can notably improve the mechanical properties and promote cell adhesion, growth, and differentiation. Thus, the colloidal gold-collagen nanoconjugates synthesized by such a straightforward and clean manner, analogous to a biomineralization pathway, provide new alternatives for developing biologically based hybrid biomaterials toward a range of therapeutic and diagnostic applications.

  6. Modeling and computations of the intramolecular electron transfer process in the two-heme protein cytochrome c4

    DEFF Research Database (Denmark)

    Natzmutdinov, Renat R.; Bronshtein, Michael D.; Zinkicheva, Tamara T.;


    The di-heme protein Pseudomonas stutzeri cytochrome c4 (cyt c4) has emerged as a useful model for studying long-range protein electron transfer (ET). Recent experimental observations have shown a dramatically different pattern of intramolecular ET between the two heme groups in different local...... environments. Intramolecular ET in homogeneous solution is too slow (>10 s) to be detected but fast (ms–μs) intramolecular ET in an electrochemical environment has recently been achieved by controlling the molecular orientation of the protein assembled on a gold electrode surface. In this work we have...

  7. Synthesis of Crown Ether-tethered β-Cyclodextrin and Fabrication of Its Self-assembled Monolayer on Gold Surface

    Institute of Scientific and Technical Information of China (English)


    A novel β-cyclodextrin derivative 6 bearing a crown ether moiety has been synthesized by a convenient method in 9.4% yield. Its self-assembled monolayer (SAM) was fabricated on the gold surface, which was characterized by using surface-enhanced Raman spectra.

  8. Synthesis and sonication-induced assembly of Si-DDR particles for close-packed oriented layers. (United States)

    Kim, Eunjoo; Cai, Wanxi; Baik, Hionsuck; Nam, Jaewook; Choi, Jungkyu


    Here, we report a seeded growth protocol for synthesizing monodisperse Si-DDR particles of ~1.3-10 μm by varying the seed amount. These Si-DDR particles were deposited onto porous α-Al2O3 discs via sonication-induced assembly, constituting close-packed h0h-oriented layers.

  9. A novel perylene diimide-based tetrahedral molecule: Synthesis, characterization and self-assembly with gold nanoparticles

    Indian Academy of Sciences (India)

    Jun-bo Li; Xiang-Lin Yu; Jing Fu; Xiwen Liu; Yang Zeng


    In this study, a novel tetrahedral molecule TPPY was successfully designed and synthesized. The self-assembly of TPPY with gold nanoparticles (Au NPs) in toluene has also been investigated. The aggregation morphologies of Au NPs can be controlled to produce different aggregate structures by changing the concentration of ligand TPPY.

  10. Heme oxygenase-1, oxidation, inflammation and atherosclerosis

    Directory of Open Access Journals (Sweden)

    Jesus A Araujo


    Full Text Available Atherosclerosis is an inflammatory process of the vascular wall characterized by the infiltration of lipids and inflammatory cells. Oxidative modifications of infiltrating low density lipoproteins and induction of oxidative stress play a major role in lipid retention in the vascular wall, uptake by macrophages and generation of foam cells, a hallmark of this disorder. The vasculature has a plethora of protective resources against oxidation and inflammation, many of them regulated by the Nrf2 transcription factor. Heme oxygenase-1 (HO-1 is a Nrf2-regulated gene that plays a critical role in the prevention of vascular inflammation. It is the inducible isoform of heme oxygenase, responsible for the oxidative cleavage of heme groups leading to the generation of biliverdin, carbon monoxide and release of ferrous iron. HO-1 has important antioxidant, antiinflammatory, antiapoptotic, antiproliferative and immunomodulatory effects in vascular cells, most of which play a significant role in the protection against atherogenesis. HO-1 may also be an important feature in macrophage differentiation and polarization to certain subtypes. The biological effects of HO-1 are largely attributable to its enzymatic activity, which can be conceived as a system with three arms of action, corresponding to its three enzymatic byproducts. HO-1 mediated vascular protection may be due to a combination of systemic and vascular local effects. It is usually expressed at low levels but can be highly upregulated in the presence of several proatherogenic stimuli. The HO-1 system is amenable for use in the development of new therapies, some of them currently under experimental and clinical trials. Interestingly, in contrast to the HO-1 antiatherogenic actions, the expression of its transcriptional regulator Nrf2 leads to proatherogenic effects instead. This article reviews the evidence that supports the antiatherogenic role of HO-1, potential pathways and mechanisms mediating

  11. Targeting heme oxygenase-1 in vascular disease. (United States)

    Durante, William


    Heme oxygenase-1 (HO-1) metabolizes heme to generate carbon monoxide (CO), biliverdin, and iron. Biliverdin is subsequently metabolized to bilirubin by biliverdin reductase. HO-1 has recently emerged as a promising therapeutic target in the treatment of vascular disease. Pharmacological induction or gene transfer of HO-1 ameliorates vascular dysfunction in animal models of atherosclerosis, post-angioplasty restenosis, vein graft stenosis, thrombosis, myocardial infarction, and hypertension, while inhibition of HO-1 activity or gene deletion exacerbates these disorders. The vasoprotection afforded by HO-1 is largely attributable to its end products: CO and the bile pigments, biliverdin and bilirubin. These end products exert potent anti-inflammatory, antioxidant, anti-apoptotic, and anti-thrombotic actions. In addition, CO and bile pigments act to preserve vascular homeostasis at sites of arterial injury by influencing the proliferation, migration, and adhesion of vascular smooth muscle cells, endothelial cells, endothelial progenitor cells, or leukocytes. Several strategies are currently being developed to target HO-1 in vascular disease. Pharmacological induction of HO-1 by heme derivatives, dietary antioxidants, or currently available drugs, is a promising near-term approach, while HO-1 gene delivery is a long-term therapeutic goal. Direct administration of CO via inhalation or through the use of CO-releasing molecules and/or CO-sensitizing agents provides an attractive alternative approach in targeting HO-1. Furthermore, delivery of bile pigments, either alone or in combination with CO, presents another avenue for protecting against vascular disease. Since HO-1 and its products are potentially toxic, a major challenge will be to devise clinically effective therapeutic modalities that target HO-1 without causing any adverse effects.

  12. Bone marrow: its contribution to heme catabolism. (United States)

    Mähönen, Y; Anttinen, M; Vuopio, P; Tenhunen, R


    Heme oxygenase (HO) and biliverdin reductase (BR), the two NADPH-dependent enzymes involved in the degradation of hemoglobin and its derivatives, were measured in bone marrow aspirates from 5 hematologically normal persons, 4 patients with chronic leucemia (CL), 11 patients with acute leucemia (AL), 8 patients with refractory sideroblastic anemia (RA), 7 patients with iron-deficiency anemia (IA), 5 patients with hemolytic anemia (HA), and 7 patients with secondary anemia (SA) to determine the enzymatic capacity of the bone marrow in different hematologic disorders for heme catabolism. HO activity in the bone marrow of normal persons was 0.42 +/- 0.28 (SD) nmoles bilirubin/10 mg protein/min; in CL, 2.15 +/- 1.34; in AL, 0.39 +/- 0.25; in RA, 0.58 +/- 0.37; in IA, 0.41 +/- 0.28; in HA, 2.56 +/- 1.40; and in SA, 1.72 +/- 1.06. BR activity, respectively, was in normal persons 8.7 +/- 2.4 (SD) nmoles bilirubin/10 mg protein/min; in CL, 13.6 +/- 9.1; in AL, 3.8 +/- 3.1 in RA, 5.1 +/- 2.7; in IA, 5.5 +/- 3.7; in HA, 17.0 +/- 7.2; and in SA, 10.5 +/- 4.2. On the basis of these findings it seems evident that both oxygenase and biliverdin reductase activities of the bone marrow are capable of adaptive regulation. The physiologic role of bone marrow in heme catabolism seems to be of significant importance.

  13. Gas-phase spectroscopy of ferric heme-NO complexes

    DEFF Research Database (Denmark)

    Wyer, J.A.; Jørgensen, Anders; Pedersen, Bjarke;


    Weakly bound complexes between ferric heme cations and NO were synthesised in the gas phase from ion-molecule reactions, and their absorption measured based on photodissociation yields. The Soret band, which serves as an important marker band for heme-protein spectroscopy, is maximal at 357±5 nm...

  14. Heme oxygenase-1 and carbon monoxide in pulmonary medicine

    NARCIS (Netherlands)

    Slebos, DJ; Ryter, SW; Choi, AMK


    Heme oxygenase-1 (HO-1), an inducible stress protein, confers cytoprotection against oxidative stress in vitro and in vivo. In addition to its physiological role in heme degradation, HO-1 may influence a number of cellular processes, including growth, inflammation, and apoptosis. By virtue of anti-i

  15. Spectroscopy of Ferric Heme and Protoporphyrin IX Ions In Vacuo

    DEFF Research Database (Denmark)

    Wyer, Jean; Nielsen, Steen Brøndsted


    This chapter deals with gas-phase spectroscopy of protoporphyrin IX and heme ions, two important biochromophores in nature. These ions strongly absorb blue and green light, which accounts for e.g. the red colour of blood. We present absorption spectra of four-coordinate ferric heme cations at room...

  16. Hemolysis-induced lethality involves inflammasome activation by heme. (United States)

    Dutra, Fabianno F; Alves, Letícia S; Rodrigues, Danielle; Fernandez, Patricia L; de Oliveira, Rosane B; Golenbock, Douglas T; Zamboni, Dario S; Bozza, Marcelo T


    The increase of extracellular heme is a hallmark of hemolysis or extensive cell damage. Heme has prooxidant, cytotoxic, and inflammatory effects, playing a central role in the pathogenesis of malaria, sepsis, and sickle cell disease. However, the mechanisms by which heme is sensed by innate immune cells contributing to these diseases are not fully characterized. We found that heme, but not porphyrins without iron, activated LPS-primed macrophages promoting the processing of IL-1β dependent on nucleotide-binding domain and leucine rich repeat containing family, pyrin domain containing 3 (NLRP3). The activation of NLRP3 by heme required spleen tyrosine kinase, NADPH oxidase-2, mitochondrial reactive oxygen species, and K(+) efflux, whereas it was independent of heme internalization, lysosomal damage, ATP release, the purinergic receptor P2X7, and cell death. Importantly, our results indicated the participation of macrophages, NLRP3 inflammasome components, and IL-1R in the lethality caused by sterile hemolysis. Thus, understanding the molecular pathways affected by heme in innate immune cells might prove useful to identify new therapeutic targets for diseases that have heme release.

  17. Engineering Non-Heme Mono- and Dioxygenases for Biocatalysis

    Directory of Open Access Journals (Sweden)

    Adi Dror


    Full Text Available Oxygenases are ubiquitous enzymes that catalyze the introduction of one or two oxygen atoms to unreactive chemical compounds. They require reduction equivalents from NADH or NADPH and comprise metal ions, metal ion complexes, or coenzymes in their active site. Thus, for industrial purposes, oxygenases are most commonly employed using whole cell catalysis, to alleviate the need for co-factor regeneration. Biotechnological applications include bioremediation, chiral synthesis, biosensors, fine chemicals, biofuels, pharmaceuticals, food ingredients and polymers. Controlling activity and selectivity of oxygenases is therefore of great importance and of growing interest to the scientific community. This review focuses on protein engineering of non-heme monooxygenases and dioxygenases for generating improved or novel functionalities. Rational mutagenesis based on x-ray structures and sequence alignment, as well as random methods such as directed evolution, have been utilized. It is concluded that knowledge-based protein engineering accompanied with targeted libraries, is most efficient for the design and tuning of biocatalysts towards novel substrates and enhanced catalytic activity while minimizing the screening efforts.

  18. Organic-inorganic random copolymers from methacrylate-terminated poly(ethylene oxide) with 3-methacryloxypropylheptaphenyl polyhedral oligomeric silsesquioxane: synthesis via RAFT polymerization and self-assembly behavior. (United States)

    Wei, Kun; Li, Lei; Zheng, Sixun; Wang, Ge; Liang, Qi


    In this contribution, we report the synthesis of organic-inorganic random polymers from methacrylate-terminated poly(ethylene oxide) (MAPEO) (Mn = 950) and 3-methacryloxypropylheptaphenyl polyhedral oligomeric silsesquioxane (MAPOSS) macromers via reversible addition-fragmentation chain transfer (RAFT) polymerization with 4-cyano-4-(thiobenzoylthio) valeric acid (CTBTVA) as the chain transfer agent. The organic-inorganic random copolymers were characterized by means of (1)H NMR spectroscopy, gel permeation chromatography (GPC) and differential scanning calorimetry (DSC). The results of GPC indicate that the polymerizations were carried out in a controlled fashion. Transmission electron microscopy (TEM) showed that the organic-inorganic random copolymers in bulk were microphase-separated and the POSS microdomains were formed via POSS-POSS interactions. In aqueous solutions the organic-inorganic random copolymers were capable of self-assembling into spherical nanoobjects as evidenced by transmission electron microscopy (TEM) and dynamic laser scattering (DLS). The self-assembly behavior of the organic-inorganic random copolymers was also found to occur in the mixtures with the precursors of epoxy. The nanostructures were further fixed via subsequent curing reaction and thus the organic-inorganic nanocomposites were obtained. The formation of nanophases in epoxy thermosets was confirmed by transmission electron microscopy (TEM) and dynamic mechanical thermal analysis (DMTA). The organic-inorganic nanocomposites displayed the enhanced surface hydrophobicity as evidenced by surface contact angle measurements.

  19. Ethylene Glycol Intercalated Cobalt/Nickel Layered Double Hydroxide Nanosheet Assemblies with Ultrahigh Specific Capacitance: Structural Design and Green Synthesis for Advanced Electrochemical Storage. (United States)

    Wang, Changhui; Zhang, Xiong; Xu, Zhongtang; Sun, Xianzhong; Ma, Yanwei


    Because of the rapid depletion of fossil fuels and severe environmental pollution, more advanced energy-storage systems need to possess dramatically improved performance and be produced on a large scale with high efficiency while maintaining low-enough costs to ensure the higher and wider requirements. A facile, energy-saving process was successfully adopted for the synthesis of ethylene glycol intercalated cobalt/nickel layered double hydroxide (EG-Co/Ni LDH) nanosheet assembly variants with higher interlayer distance and tunable transitional-metal composition. At an optimized starting Co/Ni ratio of 1, the nanosheet assemblies display a three-dimensional, spongelike network, affording a high specific surface area with advantageous mesopore structure in 2-5 nm containing large numbers of about 1.2 nm micropores for promoting electrochemical reaction. An unprecedented electrochemical performance was achieved, with a specific capacitance of 4160 F g(-1) at a discharge current density of 1 A g(-1) and of 1313 F g(-1) even at 50 A g(-1), as well as excellent cycling ability. The design and optimization of EG-Co/Ni LDH nanosheets in compositions, structures, and performances, in conjunction with the easy and relatively "green" synthetic process, will play a pivotal role in meeting the needs of large-scale manufacture and widespread application for advanced electrochemical storage.

  20. Synthesis and controlled self-assembly of covalently linked hexa-peri-hexabenzocoronene/perylene diimide dyads as models to study fundamental energy and electron transfer processes. (United States)

    Dössel, Lukas F; Kamm, Valentin; Howard, Ian A; Laquai, Frédéric; Pisula, Wojciech; Feng, Xinliang; Li, Chen; Takase, Masayoshi; Kudernac, Tibor; De Feyter, Steven; Müllen, Klaus


    We report the synthesis and photophysical characterization of a series of hexa-peri-hexabenzocoronene (HBC)/perylenetetracarboxy diimide (PDI) dyads that are covalently linked with a rigid bridge. Both the ratio of the two components and the conjugation of the bridging element are systematically modified to study the influence on self-assembly and energy and electron transfer between electron donor HBC and acceptor PDI. STM and 2D-WAXS experiments reveal that both in solution and in bulk solid state the dyads assemble into well-ordered two-dimensional supramolecular structures with controllable mutual orientations and distances between donor and acceptor at a nanoscopic scale. Depending on the symmetry of the dyads, either columns with nanosegregated stacks of HBC and PDI or interdigitating networks with alternating HBC and PDI moieties are observed. UV-vis, photoluminescence, transient photoluminescence, and transient absorption spectroscopy confirm that after photoexcitation of the donor HBC a photoinduced electron transfer between HBC and PDI can only compete with the dominant Förster resonance energy transfer, if facilitated by an intimate stacking of HBC and PDI with sufficient orbital overlap. However, while the alternating stacks allow efficient electron transfer, only the nanosegregated stacks provide charge transport channels in bulk state that are a prerequisite for application as active components in thin film electronic devices. These results have important implications for the further design of functional donor-acceptor dyads, being promising materials for organic bulk heterojunction solar cells and field-effect transistors.

  1. Extracellular heme uptake and the challenges of bacterial cell membranes. (United States)

    Smith, Aaron D; Wilks, Angela


    In bacteria, the fine balance of maintaining adequate iron levels while preventing the deleterious effects of excess iron has led to the evolution of sophisticated cellular mechanisms to obtain, store, and regulate iron. Iron uptake provides a significant challenge given its limited bioavailability and need to be transported across the bacterial cell wall and membranes. Pathogenic bacteria have circumvented the iron-availability issue by utilizing the hosts' heme-containing proteins as a source of iron. Once internalized, iron is liberated from the porphyrin enzymatically for cellular processes within the bacterial cell. Heme, a lipophilic and toxic molecule, poses a significant challenge in terms of transport given its chemical reactivity. As such, pathogenic bacteria have evolved sophisticated membrane transporters to coordinate, sequester, and transport heme. Recent advances in the biochemical and structural characterization of the membrane-bound heme transport proteins are discussed in the context of ligand coordination, protein-protein interaction, and heme transfer.

  2. Caenorhabditis elegans ATAD-3 modulates mitochondrial iron and heme homeostasis. (United States)

    van den Ecker, Daniela; Hoffmann, Michael; Müting, Gesine; Maglioni, Silvia; Herebian, Diran; Mayatepek, Ertan; Ventura, Natascia; Distelmaier, Felix


    ATAD3 (ATPase family AAA domain-containing protein 3) is a mitochondrial protein, which is essential for cell viability and organismal development. ATAD3 has been implicated in several important cellular processes such as apoptosis regulation, respiratory chain function and steroid hormone biosynthesis. Moreover, altered expression of ATAD3 has been associated with several types of cancer. However, the exact mechanisms underlying ATAD3 effects on cellular metabolism remain largely unclear. Here, we demonstrate that Caenorhabditis elegans ATAD-3 is involved in mitochondrial iron and heme homeostasis. Knockdown of atad-3 caused mitochondrial iron- and heme accumulation. This was paralleled by changes in the expression levels of several iron- and heme-regulatory genes as well as an increased heme uptake. In conclusion, our data indicate a regulatory role of C. elegans ATAD-3 in mitochondrial iron and heme metabolism.

  3. Multi-heme proteins: nature's electronic multi-purpose tool. (United States)

    Bewley, Kathryn D; Ellis, Katie E; Firer-Sherwood, Mackenzie A; Elliott, Sean J


    While iron is often a limiting nutrient to Biology, when the element is found in the form of heme cofactors (iron protoporphyrin IX), living systems have excelled at modifying and tailoring the chemistry of the metal. In the context of proteins and enzymes, heme cofactors are increasingly found in stoichiometries greater than one, where a single protein macromolecule contains more than one heme unit. When paired or coupled together, these protein associated heme groups perform a wide variety of tasks, such as redox communication, long range electron transfer and storage of reducing/oxidizing equivalents. Here, we review recent advances in the field of multi-heme proteins, focusing on emergent properties of these complex redox proteins, and strategies found in Nature where such proteins appear to be modular and essential components of larger biochemical pathways. This article is part of a Special Issue entitled: Metals in Bioenergetics and Biomimetics Systems.

  4. Synthesis of Janus-like gold nanoparticles with hydrophilic/hydrophobic faces by surface ligand exchange and their self-assemblies in water. (United States)

    Iida, Ryo; Kawamura, Hitoshi; Niikura, Kenichi; Kimura, Takashi; Sekiguchi, Shota; Joti, Yasumasa; Bessho, Yoshitaka; Mitomo, Hideyuki; Nishino, Yoshinori; Ijiro, Kuniharu


    This study aims at the synthesis of Janus gold nanoparticles (Janus GNPs) with hydrophilic/hydrophobic faces by a simple ligand exchange reaction in an homogeneous system and at the elucidation of the self-assembled structures of the Janus GNPs in water. As hydrophilic surface ligands, we synthesized hexaethylene glycol (E6)-terminated thiolate ligands with C3, C7, or C11 alkyl chains, referred to as E6C3, E6C7, and E6C11, respectively. As a hydrophobic ligand, a butyl-headed thiolate ligand C4-E6C11, in which a C4 alkyl was introduced on the E6C11 terminus, was synthesized. The degree of segregation between the two ligands on the GNPs (5 nm in diameter) was examined by matrix-assisted laser desorption/ionization time-of fright mass spectrometry (MALDI-TOF MS) analysis. We found that the choice of immobilization methods, one-step or two-step addition of the two ligands to the GNP solution, crucially affects the degree of segregation. The two-step addition of a hydrophilic ligand (E6C3) followed by a hydrophobic ligand (C4-E6C11) produced a large degree of segregation on the GNPs, providing Janus-like GNPs. When dispersed in water, these Janus-like GNPs formed assemblies of ∼160 nm in diameter, whereas Domain GNPs, in which the two ligands formed partial domains on the surface, were precipitated even when the molar ratio of the hydrophilic ligand and the hydrophobic ligand on the surface of the NPs was almost 1:1. The assembled structure of the Janus-like GNPs in water was directly observed by pulsed coherent X-ray solution scattering using an X-ray free-electron laser, revealing irregular spherical structures with uneven surfaces.


    Institute of Scientific and Technical Information of China (English)

    Huai-chao Wang; Ming-zu Zhang; Pei-hong Ni; Jin-lin He; Ying Hao; Yi-xian Wu


    Two pH-responsive amphiphilic diblock copolymers,namely polyisobutylene-block-poly[2-(N,N-dimethylamino)ethyl methacrylate] (PIB-b-PDMAEMA) and polyisobutylene-block-poly(metharylic acid) (PIB-b-PMAA),were synthesized via oxyanion-initiated polymerization,and their multiple self-assembly behaviors have been studied.An exo-o1efin-terminated highly reactive polyisobutylene (HRPIB) was first changed to hydroxyl-terminated PIB (PIB-OH) via hydroboration-oxidation of C=C double bond in the chain end,and then reacted with KH to yield a potassium alcoholate of PIB (PIB-O-K+).PIB-O-K+ was immediately used as a macroinitiator to polymerize DMAEMA monomer,resulting in a cationic diblock copolymer PIB-b-PDMAEMA.With the similar synthesis procedure,the anionic diblock copolymer PIB-b-PMAA could be prepared via a combination of oxyanion-initiated polymerization of tert-butyl methacrylate (tBMA) and subsequent hydrolysis of tert-butyl ester groups in PtBMA block.The functional PIB and block copolymers have been fully characterized by 1H-NMR,FT-IR spectroscopy,and gel permeation chromatography (GPC).These samples allowed us to systematically investigate the effects of block composition on the pH responsivity and various self-assembled morphologies of the copolymers in THF/water mixed solvent.Transmission electron microscopy (TEM) images revealed that these diblock copolymers containing small amount of original PIB without exo-olefin-terminated group are able to self-assemble into micelles,vesicles with different particle sizes and cylindrical aggregates,depending on various factors including block copolymer composition,solvent polarity and pH value.

  6. One-step self-assembled synthesis of CuO with tunable hierarchical structures and their electrocatalytic properties for nitrite oxidation in aqueous media. (United States)

    Zhao, Youcheng; Song, Xinyu; Yin, Zhilei; Song, Qisheng


    Controlled synthesis of CuO with various hierarchical structures consisting of self-organized nanoparticles is realized by using n-octylamine (OLA) as a structure inducing agent via a facile hydrothermal synthetic method. The growth and assemblage of CuO can be finely tuned by selecting the preparative parameters. In particular, it is found that the degree of the hierarchical organization can be modulated by simply changing the amount of the n-octylamine and CuO nanoparticles exhibit self-assembled two-dimensional (2D) sheet-like, three-dimensional (3D) disk-like and bowknot-like architectures, respectively. In the present case, OLA serves as a capping surfactant that can modulate growth of CuO nanocrystals via hydrophobic forces between the OLA molecules. CuO nanoparticles can be self-assembled into different complex architectures depending on the strength of hydrophobic forces. Hierarchical sphere-like CuO assembled from nanorods can also be easily fabricated by adjusting the starting NaOH to CuCl2 volume ratio, in which OLA serves not only as the structure-directing agent, but also as a weak base agent to produce hydroxyl anions. The electrochemical performances of the as-synthesized different products for sensing nitrite oxidation are evaluated. The results reveal that the electrocatalytic activity is related to the secondary nanostructures. Compared to the others, the bowknot-shaped and sphere-shaped CuO products exhibit excellent electrocatalytic activity toward nitrite oxidation and fast current response in nitrite sensing because of their peculiar hierarchical structures with high BET surface areas and well-ordered pores.

  7. Evaluation of unbound free heme in plant cells by differential acetone extraction. (United States)

    Espinas, Nino A; Kobayashi, Koichi; Takahashi, Shigekazu; Mochizuki, Nobuyoshi; Masuda, Tatsuru


    Heme functions not only as a prosthetic group of hemoproteins but also as a regulatory molecule, suggesting the presence of 'free' heme. Classically, total non-covalently bound heme is extracted from plant samples with acidic acetone after removal of pigments with basic and neutral acetone. Earlier work proposed that free heme can be selectively extracted into basic acetone. Using authentic hemoproteins, we confirmed that acidic acetone can quantitatively extract heme, while no heme was extracted into neutral acetone. Meanwhile, a certain amount of heme was extracted into basic acetone from hemoglobin and myoglobin. Moreover, basic acetone extracted loosely bound heme from bovine serum albumin, implying that the nature of hemoproteins largely influences heme extraction into basic acetone. Using a highly sensitive heme assay, we found that basic and neutral acetone can extract low levels of heme from plant samples. In addition, neutral acetone quantitatively extracted free heme when it was externally added to plant homogenates. Furthermore, the level of neutral acetone-extractable heme remained unchanged by precursor (5-aminolevulinic acid) feeding, while increased by norflurazon treatment which abolishes chloroplast biogenesis. However, changes in these heme levels did not correlate to genomes uncoupled phenotypes, suggesting that the level of unbound free heme would not affect retrograde signaling from plastids to the nucleus. The present data demonstrate that the combination of single-step acetone extraction following a sensitive heme assay is the ideal method for determining total and free heme in plants.

  8. Synthesis of a tetracyclic G∧C scaffold for the assembly of rosette nanotubes with 1.7 nm inner diameter. (United States)

    Borzsonyi, Gabor; Alsbaiee, Alaaeddin; Beingessner, Rachel L; Fenniri, Hicham


    The synthesis of a tetracyclic self-complementary molecule 4 for self-assembly into rosette nanotubes is presented. This new heterocycle has a core structure containing two pyrido[2,3-d]pyrimidine molecules fused together and features the Watson-Crick hydrogen bond donor-acceptor arrays of both guanine (G) and cytosine (C). Current methods to synthesize pyrido[2,3-d]pyrimidines require harsh conditions and long reaction times and result usually in low product yields. This is particularly problematic for the direct incorporation of functional groups that cannot withstand these conditions. Here, we present an efficient approach to access the multifunctional pyrido[2,3-d]pyrimidine intermediate 2 under relatively mild conditions using three regioselective S(N)Ar reactions at C2, C4, and C7 on the trichloro compound 1. The electron-withdrawing group and amino functionalities on 2 are then used as a handle to install the third and fourth rings of 4 using a Friedländer-type condensation followed by mixed urea synthesis and cyclization.

  9. Synthesis of nanoparticle/ligand composite thin films by sequential ligand self assembly and surface complex reduction. (United States)

    Muench, Falk; Fuchs, Anne; Mankel, Eric; Rauber, Markus; Lauterbach, Stefan; Kleebe, Hans-Joachim; Ensinger, Wolfgang


    Nanocomposite thin films consisting of ligand-connected metal nanoparticles were deposited by iteration of ligand assembly, surface complex formation and reduction. This novel and convenient approach combines characteristics of the layer-by-layer (LbL) and the successive ion layer adsorption and reaction (SILAR) techniques. In contrast to classical LbL assembly, the nanoparticle formation is performed in situ, avoiding separate reduction, protection and attachment steps. To demonstrate the versatility of the approach, different metal precursors (Pd, Ag and Au salts) and linkers (1,2-ethanedithiol, 1,4-benzenedithiol and polythiol) were applied. The formation of dithiol-linked nanoparticle films was confirmed by TEM and XPS. By combining the deposition protocol with ion track etched polycarbonate templates, nanotubes and nanowires with high aspect ratios of up to 300 could be fabricated.

  10. From Polymeric Nanoparticles to Dye-containing Photonic Crystals:Synthesis,Self-assembling,Optical Features, Possible Applications

    Institute of Scientific and Technical Information of China (English)

    A.V.Yakimansky; A.Yu.Menshikova; N.N.Shevchenko; A.G.Bazhenova; S.K.Sazonov; A.I.Vedernikov; S.P.Gromov; V.A.Sazhnikov; M.V.Alfimov


    1 Results Self-assembling of monodisperse polymeric nanoparticles is a perspective method of obtaining photonic crystalline materials for optoelectronics,telecommunication industry and optosensorics.For tuning optical characteristics of photonic crystals it is advisable to functionalize nanoparticles by dyes absorbing or emitting light in the vicinity of the photonic band gap,which position depends on the nanoparticle diameter.To prepare monodisperse nanoparticles with the dye-functionalyzed surface emu...

  11. Alternating donor-acceptor arrays from hexa-peri-hexabenzocoronene and benzothiadiazole: synthesis, optical properties, and self-assembly. (United States)

    Hinkel, Felix; Cho, Don; Pisula, Wojciech; Baumgarten, Martin; Müllen, Klaus


    Donor-acceptor (D-A) structures were obtained by alternating arrays of hexa-peri-hexabenzocoronene (HBC) and benzo[c][1,2,5]thiadiazole (BTZ). Optoelectronic investigations revealed a charge transfer due to strong push-pull interactions. 2 D wide-angle X-ray scattering (WAXS) data indicated an arrangement in liquid-crystalline columnar assemblies, in which the π-stacking distances and molecular orientation depend on the number of HBC units in the molecules.

  12. Synthesis of metal-hydrazone complexes and vapochromic behavior of their hydrogen-bonded proton-transfer assemblies. (United States)

    Kobayashi, Atsushi; Dosen, Masa-aki; Chang, Mee; Nakajima, Kiyohiko; Noro, Shin-ichiro; Kato, Masako


    We synthesized and investigated a new series of metal-hydrazone complexes, including deprotonated [MX(mtbhp)] and protonated forms [MX(Hmtbhp)](ClO(4)) (M = Pd(2+), Pt(2+); X = Cl(-), Br(-); Hmtbhp = 2-(2-(2-(methylthio)benzylidene)hydrazinyl)pyridine) and hydrogen-bonded proton-transfer (HBPT) assemblies containing [PdBr(mtbhp)] and bromanilic acid (H(2)BA). The mtbhp hydrazone ligand acts as a tridentate SNN ligand and provides a high proton affinity. UV-vis spectroscopy revealed that these metal-hydrazone complexes follow a reversible protonation-deprotonation reaction ([MX(mtbhp)] + H(+) ⇋ [MX(Hmtbhp)](+)), resulting in a remarkable color change from red to yellow. Reactions between proton acceptor [PdBr(mtbhp)] (A) and proton donor H(2)BA (D) afforded four types of HBPT assemblies with different D/A ratios: for D/A = 1:1, {[PdBr(Hmtbhp)](HBA)·Acetone} and {[PdBr(Hmtbhp)](HBA)·2(1,4-dioxane)}; for D/A = 1:2, [PdBr(Hmtbhp)](2)(BA); and for D/A = 3:2, {[PdBr(Hmtbhp)](2)(HBA)(2)(H(2)BA)·2Acetonitrile}. The proton donor gave at least one proton to the acceptor to form the hydrogen bonded A···D pair of [PdBr(Hmtbhp)](+)···HBA(-). The strength of the hydrogen bond in the pair depends on the kind of molecule bound to the free monoanionic bromanilate OH group. Low-temperature IR spectra (T < 150 K) showed that the hydrogen bond distance between [PdBr(Hmtbhp)](+) and bromanilate was short enough (ca. 2.58 Å) to induce proton migration in the [PdBr(Hmtbhp)](2)(BA) assembly in the solid state. The hydrogen bonds formed not only between [PdBr(Hmtbhp)](+) and HBA(-) but also between HBA(-) and neutral H(2)BA molecules in the {[PdBr(Hmtbhp)](2)(HBA)(2)(H(2)BA)·2Acetonitrile} assembly. The H(2)BA-based flexible hydrogen bond network and strong acidic host structure result in an interesting vapor adsorption ability and vapochromic behavior in this assembly because the vapor-induced rearrangement of the hydrogen bond network, accompanied by changes in

  13. Effect of Promoters and Plasmid Copy Number on Cyt1A Synthesis and Crystal Assembly in Bacillus thuringiensis. (United States)

    Park, Hyun-Woo; Hice, Robert H; Federici, Brian A


    Cyt1Aa is a major mosquitocidal protein synthesized during sporulation of Bacillus thuringiensis subsp. israelensis, composing more than 50% of its parasporal body. This high level of synthesis is due to several factors including three strong sporulation-dependent promoters, a strong transcription termination sequence, and an associated 20-kDa helper protein. Cyt1Aa's toxicity is low compared to the Cry proteins of this species, namely, Cry4Aa, Cry4Ba, and Cry11Aa, but it nevertheless plays an important role in the biology of B. thuringiensis subsp. israelensis in that it synergizes their mosquitocidal toxicity and suppresses the evolution of resistance. In the present study, the effects of using different cyt1Aa promoter combinations and plasmid copy number on synthesis of Cyt1Aa were evaluated. Using the 4Q7 (plasmid-cured) strain of B. thuringiensis subsp. israelensis as an experimental host, a plasmid copy number of two or three yielded no Cyt1Aa, whereas a copy number of four yielded only small crystals, even when expression was driven by one of the wild-type promoters. However, using all three wild-type promoters and a plasmid copy number of 20 yielded Cyt1A crystals tenfold larger than those produced by one promoter and a plasmid copy number of four. High levels of Cyt1Aa synthesis resulted in significantly fewer spores per unit medium and imperfectly formed crystals. Similar results were obtained when Cyt1Aa synthesis was evaluated using the same expression constructs in a mutant strain of B. thuringiensis subsp. israelensis that lacks the cyt1Aa gene.

  14. Electrochemical determination of heme-linked pKa values and the importance of using fluoride binding in heme proteins. (United States)

    Cerda, Jose F; Roeder, Margaret H; Houchins, Danielle N; Guzman, Carmen X; Amendola, Emily J; Castorino, Jacquelyn D; Fritz, Andrea L


    The ultraviolet-visible (UV-vis) spectroelectrochemical measurements of heme proteins in the presence of a heme-bound fluoride ion can be used as a probe for heme-linked ionizations of acid-base groups in the heme pocket. A detailed study of the pH dependence of the midpoint potential of skeletal horse myoglobin (Mb) with a heme-bound fluoride ion (Mb-F) reveals how protonation of the distal histidine (H64) changes the redox properties of the protein with a determined pKa of 5.3. In addition, fluoride binding in myoglobin provides a stabilization of -1.9 kcal/mol of the ferric Mb-F relative to ferric Mb without fluoride.

  15. Heme binding properties of glyceraldehyde-3-phosphate dehydrogenase. (United States)

    Hannibal, Luciana; Collins, Daniel; Brassard, Julie; Chakravarti, Ritu; Vempati, Rajesh; Dorlet, Pierre; Santolini, Jérôme; Dawson, John H; Stuehr, Dennis J


    Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a glycolytic enzyme that also functions in transcriptional regulation, oxidative stress, vesicular trafficking, and apoptosis. Because GAPDH is required for the insertion of cellular heme into inducible nitric oxide synthase [Chakravarti, R., et al. (2010) Proc. Natl. Acad. Sci. U.S.A. 107, 18004-18009], we extensively characterized the heme binding properties of GAPDH. Substoichiometric amounts of ferric heme bound to GAPDH (one heme per GAPDH tetramer) to form a low-spin complex with UV-visible maxima at 362, 418, and 537 nm and when reduced to ferrous gave maxima at 424, 527, and 559 nm. Ferric heme association and dissociation rate constants at 10 °C were as follows: k(on) = 17800 M(-1) s(-1), k(off1) = 7.0 × 10(-3) s(-1), and k(off2) = 3.3 × 10(-4) s(-1) (giving approximate affinities of 19-390 nM). Ferrous heme bound more poorly to GAPDH and dissociated with a k(off) of 4.2 × 10(-3) s(-1). Magnetic circular dichroism, resonance Raman, and electron paramagnetic resonance spectroscopic data on the ferric, ferrous, and ferrous-CO complexes of GAPDH showed that the heme is bis-ligated with His as the proximal ligand. The distal ligand in the ferric complex was not displaced by CN(-) or N(3)(-) but in the ferrous complex could be displaced by CO at a rate of 1.75 s(-1) (for >0.2 mM CO). Studies with heme analogues revealed selectivity toward the coordinating metal and porphyrin ring structure. The GAPDH-heme complex was isolated from bacteria induced to express rabbit GAPDH in the presence of δ-aminolevulinic acid. Our finding of heme binding to GAPDH expands the protein's potential roles. The strength, selectivity, reversibility, and redox sensitivity of heme binding to GAPDH are consistent with it performing heme sensing or heme chaperone-like functions in cells.

  16. Increase on the initial soluble heme levels in acidic conditions is an important mechanism for spontaneous heme crystallization in vitro.

    Directory of Open Access Journals (Sweden)

    Renata Stiebler

    Full Text Available BACKGROUND: Hemozoin (Hz is a heme crystal that represents a vital pathway for heme disposal in several blood-feeding organisms. Recent evidence demonstrated that β-hematin (βH (the synthetic counterpart of Hz formation occurs under physiological conditions near synthetic or biological hydrophilic-hydrophobic interfaces. This seems to require a heme dimer acting as a precursor of Hz crystals that would be formed spontaneously in the absence of the competing water molecules bound to the heme iron. Here, we aimed to investigate the role of medium polarity on spontaneous βH formation in vitro. METHODOLOGY/PRINCIPAL FINDINGS: We assessed the effect of water content on spontaneous βH formation by using the aprotic solvent dimethylsulfoxide (DMSO and a series of polyethyleneglycols (PEGs. We observed that both DMSO and PEGs (3.350, 6.000, 8.000, and 22.000 increased the levels of soluble heme under acidic conditions. These compounds were able to stimulate the production of βH crystals in the absence of any biological sample. Interestingly, the effects of DMSO and PEGs on βH formation were positively correlated with their capacity to promote previous heme solubilization in acidic conditions. Curiously, a short chain polyethyleneglycol (PEG 300 caused a significant reduction in both soluble heme levels and βH formation. Finally, both heme solubilization and βH formation strongly correlated with reduced medium water activity provided by increased DMSO concentrations. CONCLUSIONS: The data presented here support the notion that reduction of the water activity is an important mechanism to support spontaneous heme crystallization, which depends on the previous increase of soluble heme levels.

  17. 氧化石墨烯的合成及组装%Synthesis and Assembling of Graphene Oxide

    Institute of Scientific and Technical Information of China (English)



    Graphene oxide was synthesized using modified Hummer’s method and assembled in solution and solid surface.The FT-IR、UV-Vis、TGA and SEM were employed to characterize the obtained Graphene oxide and its assemblies.FT-IR showed that the Graphene oxide has oxygen-containing functional groups such as hydroxy,carboxyl,epoxide and carbonyl group.UV-Vis spectra imply the influence of particle size and the pH values on the Graphene oxide dispersion.TGA measurements further confirmed the existence of these functional groups in Graphene oxide.SEM observation showed the micro morphology of the Graphene assembly from solution.%用改良的Hummer法制备了氧化石墨烯并对氧化石墨烯在溶液及玻璃表面进行了组装。采用傅里叶红外光谱(FT-IR)、热重分析(TGA)、电阻率仪、紫外可见分光光度计(UV-Vis)对氧化石墨烯及组装体系进行了表征,并用扫描电子显微镜对组装后物质的表面进行了观察。FT-IR测试表明氧化石墨烯含有羟基、羧基、环氧以及羰基等含氧官能团。UV-Vis测试表明粒径和溶剂的pH对氧化石墨烯在液相中分散性的影响。TGA测试进一步证明了氧化石墨烯含有大量含氧官能团。SEM观察揭示了石墨烯组装体的微观形貌。

  18. Synthesis of skeletally diverse alkaloid-like molecules: exploitation of metathesis substrates assembled from triplets of building blocks

    Directory of Open Access Journals (Sweden)

    Sushil K. Maurya


    Full Text Available A range of metathesis substrates was assembled from triplets of unsaturated building blocks. The approach involved the iterative attachment of a propagating and a terminating building block to a fluorous-tagged initiating building block. Metathesis cascade chemistry was used to “reprogram” the molecular scaffolds. Remarkably, in one case, a cyclopropanation reaction competed with the expected metathesis cascade process. Finally, it was demonstrated that the metathesis products could be derivatised to yield the final products. At each stage, purification was facilitated by the presence of a fluorous-tagged protecting group.


    Institute of Scientific and Technical Information of China (English)



    Dendritic poly(amidoamine)-b-poly(L-glutamate)(PAMAM-b-PLG) biohybrids were synthesized by the ring-opening polymerization ofγ-benzyl-L-glutamate N-carboxyanhydride monomer,followed by the deprotection of benzyl groups on poly(benzyl-L-glutamate),and were characterized by ~1H-NMR,FT-IR and gel permeation chromatography.The self-assembly behavior of the PAMAM-b-PLG biohybrid was investigated by means of UV-Vis,dynamic light scattering (DLS),transmission electronic microscopy(TEM) and ~1H-NMR.UV-Vis analys...

  20. Synthesis and self-assembly behavior of a biodegradable and sustainable soybean oil-based copolymer nanomicelle (United States)

    Bao, Lixia; Bian, Longchun; Zhao, Mimi; Lei, Jingxin; Wang, Jiliang


    Herein, we report a novel amphiphilic biodegradable and sustainable soybean oil-based copolymer (SBC) prepared by grafting hydrophilic and biocompatible hydroxyethyl acrylate (HEA) polymeric segments onto the natural hydrophobic soybean oil chains. FTIR, H1-NMR, and GPC measurements have been used to investigate the molecular structure of the obtained SBC macromolecules. Self-assembly behaviors of the prepared SBC in aqueous solution have also been extensively evaluated by fluorescence spectroscopy and transmission electron microscopy. The prepared SBC nanocarrier with the size range of 40 to 80 nm has a potential application in the biomedical field.

  1. Cooperative coupling and role of heme a in the proton pump of heme-copper oxidases. (United States)

    Papa, S; Capitanio, N; Villani, G; Capitanio, G; Bizzoca, A; Palese, L L; Carlino, V; De Nitto, E


    In the last few years, evidence has accumulated supporting the applicability of the cooperative model of proton pumps in cytochrome systems, vectorial Bohr mechanisms, to heme-copper oxidases. The vectorial Bohr mechanism is based on short- and long-range protonmotive cooperative effects linked to redox transitions of the metal centers. The crystal structure of oxidized and reduced bovine-heart cytochrome c oxidase reveals, upon reduction, the occurrence of long-range conformational changes in subunit I of the oxidase. Analysis of the crystal structure of cytochrome c oxidase shows the existence of hydrogen-bonded networks of amino acid residues which could undergo redox-linked pK shifts resulting in transmembrane proton translocation. Our group has identified four proteolytic groups undergoing reversible redox-linked pK shifts. Two groups result in being linked to redox transitions of heme a3. One group is apparently linked to CuB. The fourth group is linked to oxido-reduction of heme a. We have shown that the proton transfer resulting from the redox Bohr effects linked to heme a and CuB in the bovine oxidase displays membrane vectorial asymmetry, i.e., protons are taken up from the inner aqueous space (N), upon reduction, and released in the external space (P), upon oxidation of the metals. This direction of proton uptake and release is just what is expected from the vectorial Bohr mechanism. The group linked to heme a, which can transfer up to 0.9 H+/e- at pHs around neutrality, can provide the major contribution to the proton pump. It is proposed that translocation of pumped protons, linked to electron flow through heme a, utilizes a channel (channel D) which extends from a conserved aspartate at the N entrance to a conserved glutamate located between heme a and the binuclear center. The carboxylic group of this glutamic acid, after having delivered, upon electron flow through heme a, pumped protons towards the P phase, once reprotonated from the N phase, moves

  2. Synthesis of size-controlled faceted pentagonal silver nanorods with tunable plasmonic properties and self-assembly of these nanorods. (United States)

    Pietrobon, Brendan; McEachran, Matthew; Kitaev, Vladimir


    Monodisperse size-controlled faceted pentagonal silver nanorods were synthesized by thermal regrowth of decahedral silver nanoparticle (AgNPs) in aqueous solution at 95 degrees C, using citrate as a reducing agent. The width of the silver nanorods was determined by the size of the starting decahedral particle, while the length was varied from 50 nm to 2 mum by the amount of new silver added to the growth solution. Controlled regrowth allowed us to produce monodisperse AgNPs with a shape of elongated pentagonal dipyramid (regular Johnson solid, J(16)). Faceted pentagonal particles exhibited remarkable optical properties with sharp plasmon resonances precisely tunable across visible and NIR. Due to the narrow size distribution, faceted pentagonal silver nanorods readily self-assembled into the 3-D arrays similar to smectic mesophases. Hexagonal arrangement in the array completely overrode five-fold symmetry of the nanorods. Overall, our findings highlight the importance of pentagonal symmetry in metal nanoparticles and offer a facile method of the preparation of monodisperse AgNPs with controlled dimensions and plasmonic properties that are promising for optical applications and functional self-assembly.

  3. Solvothermal synthesis of magnetic Fe3O4 microparticles via self-assembly of Fe3O4 nanoparticles

    Institute of Scientific and Technical Information of China (English)

    Wei Zhang; Fenglei Shen; Ruoyu Hong


    Ferromagnetic Fe3O4 nanoparticles were synthesized and then self-assembled into microparticles via a solvothermal method, using FeCI3.6H2O as the iron source, sodium oleate as the surfactant, and ethylene glycol as the reducing agent and solvent. The obtained Fe3O4 microparticles were characterized by X-ray powder diffraction (XRD), X-ray photoelectron spectroscopy (XPS), Raman spectroscopy and vibrating sample magnetometer (VSM). The size and morphology of the particles were examined using transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The Fe3O4 microparticles of nearly monodisperse diameters, controllable in the range of 120-400 nm, consist of assemblies of Fe3O4nanoparticles with a diameter of 22 nm. The effects of reaction time, amount of surfactant and NaAc on the products were discussed. Interestingly, by using the pre-synthesized Fe3O4 microparticles as the growth substrates, spherical and smooth-looking Fe3O4 microparticles with average diameter of lμmwere obtained. A plausible formation process was discussed.

  4. Heme synthase (ferrochelatase) catalyzes the removal of iron from heme and demetalation of metalloporphyrins. (United States)

    Taketani, Shigeru; Ishigaki, Mutsumi; Mizutani, Atsushi; Uebayashi, Masashi; Numata, Masahiro; Ohgari, Yoshiko; Kitajima, Sakihito


    The red pigments in meat products, including cooked cured ham, arise from the reaction of myoglobin with nitric oxide generated from exogenous nitrite. Since carcinogenic nitrosoamines may be generated by the treatment of meats with nitrite, the production of nitrite-free meat products is an attractive alternative. Raw dry-cured (Parma) hams are produced by the treatment of meats with salts other than nitrite. Analysis of pigments in raw dry-cured hams reveals that the main pigment is zinc protoporphyrin, suggesting that the conversion of heme to zinc protoporphyrin occurs via an iron-removal reaction from myoglobin heme during the processing of raw hams. Purification of the iron-removal enzyme showed that it was identical to ferrochelatase. Recombinant ferrochelatase in combination with NADH-cytochrome b5 reductase catalyzed NADH-dependent iron-removal reaction from hemin and hemoproteins. Metal ions such as zinc and cobalt were also removed from the corresponding metalloporphyrins. The addition of zinc ions led to the formation of zinc protoporphyrin. In cultured cells, the conversion of zinc mesoporphyrin to mesoheme was observed to be dependent on ferrochelatase and could be markedly induced during erythroid differentiation. This is the first demonstration of a new enzyme reaction, the reverse reaction of ferrochelatase, which may contribute to a new route of the recycling of protoporphyrin and heme in cells.

  5. Magnetic and structural characterization of transition metal porphyrin complexes and the heme sites of heme peroxidases

    Energy Technology Data Exchange (ETDEWEB)

    Rodgers, K.R.


    Four studies of heme and heme model systems are described. The first study involves low temperature solution structural characterization of high-valent porphinatomanganese complexes via {sup 2}H- and {sup 13}C-NMR, and ESR spectroscopies. The reactive species were generated by low temperature reaction with chlorine (0) and chlorine(I) reagents. The implications of these species are discussed in terms of the relative reactivity of other +4 first row transition metal complexes and in terms of the catalytic effectiveness of porphinatomanganese (III) complexes in oxo-transfer reactions. The second study involved the analysis of isotropic {sup 2}H-NMR shifts observed for specifically deuterated chloro-N-methylporphinatomanganese(II) complexes. An ESR spectroscopic study of several ferrous heme peroxidase/NO complexes is presented. The {sup 14}N and {sup 15}N hyperfine splitting patterns and coupling constants in the ESR spectra clearly demonstrate the presence of a nitrogen-bound proximal ligand in lactoperoxidase. Finally, a catalytic autoxidation system involving cyclohexene and/or propanal as substrates is described. This reaction is catalyzed by high spin tetraarylporphinatoiron (III) complexes and evolves CO{sub 2}.

  6. Absorption by Isolated Ferric Heme Nitrosyl Cations In Vacuo

    DEFF Research Database (Denmark)

    Wyer, Jean; Nielsen, Steen Brøndsted


    Keywords:biophysics;gas-phase spectroscopy;heme proteins;mass spectrometry;nitric oxide Almost innocent: In photobiophysical studies of ferric heme nitrosyl complexes, the absorption spectra of six-coordinate complexes with NO and Met or Cys are similar to that of the five-coordinate complex ion ......(heme)(NO)+. Since the absorption spectra of related proteins with histidine as the proximal ligand are similar to those of the gaseous complexes, the protein microenvironment has little effect on the lowest-energy transition of the porphyrin macrocycle....

  7. One-Pot Synthesis of Cu2O/Cu Self-Assembled Hollow Nanospheres with Enhanced Photocatalytic Performance

    Directory of Open Access Journals (Sweden)

    Bo Zhou


    Full Text Available Cu2O/Cu hollow spheres are prepared using one-pot template-free solvent-thermal synthesis route with (CH3COO2Cu·H2O as a precursor. With the reaction time increasing gradually from 2 h to 20 h, the morphology of the Cu2O/Cu evolves from nanoparticle to hollow nanosphere. The hollow structure is obtained when the cooling rate falls down to 0.7°C/min. And the content of Cu in the hollow spheres also can be easily controlled by adjusting the solvent-thermal synthesis time. Using photocatalytic degradation of phenol as the probe molecules under visible-light illumination, we have investigated the influence of hollow structure on the photocatalytic activity of Cu2O/Cu. The prepared hollow sphere Cu2O/Cu particles exhibited a higher photodegradation capability than nanoparticles and solid spheres. When the content of Cu lies in the range of 11–86 wt%, the samples exhibit higher photocatalytic performance, indicating that the Cu2O/Cu particles with hollow structure are promising candidates for the processing of pollutants.

  8. Heme oxygenase-1-derived carbon monoxide is an autocrine inhibitor of vascular smooth muscle cell growth. (United States)

    Peyton, Kelly J; Reyna, Sylvia V; Chapman, Gary B; Ensenat, Diana; Liu, Xiao-ming; Wang, Hong; Schafer, Andrew I; Durante, William


    Vascular smooth muscle cells (SMCs) generate carbon monoxide (CO) via the catabolism of heme by the enzyme heme oxygenase (HO). In the present study, we found that serum stimulated a time- and concentration-dependent increase in the levels of HO-1 messenger RNA (mRNA) and protein in vascular SMCs. The induction of HO-1 expression by serum was inhibited by actinomycin D or cycloheximide. In addition, serum stimulated HO activity, as reflected by an increase in the concentration of bilirubin in the culture media. Treatment of vascular SMCs with serum stimulated DNA synthesis and this was potentiated by the HO inhibitors, zinc and tin protoporphyrin-IX as well as by the CO scavenger, hemoglobin. The iron chelator desferrioxamine had no effect on DNA synthesis. However, exposure of vascular SMCs to exogenous CO inhibited serum-stimulated SMC proliferation and the phosphorylation of retinoblastoma protein. In addition, CO arrested SMCs at the G(1)/S transition phase of the cell cycle and selectively blocked the serum-stimulated expression of cyclin A mRNA and protein without affecting the expression of cyclin D1 and E. CO also inhibited the serum-stimulated activation of cyclin A-associated kinase activity and cyclin-dependent kinase 2 activity. These results demonstrate that serum stimulates HO-1 gene expression and CO synthesis. Furthermore, they show that CO acts in a negative feedback fashion to inhibit vascular SMC growth by regulating specific components of the cell cycle machinery. The capacity of vascular mitogens to induce CO synthesis may provide a novel mechanism by which these agents modulate cell growth.

  9. Facile synthesis of self-assembled SnO nano-square sheets and hydrogen absorption characteristics

    Energy Technology Data Exchange (ETDEWEB)

    Iqbal, M. Zubair [Department of Physics, School of Applied Science, University of Science and Technology Beijing, Beijing 100083 (China); Wang, Fengping, E-mail: [Department of Physics, School of Applied Science, University of Science and Technology Beijing, Beijing 100083 (China); Feng, Ting [School of Metallurgical and Ecological Engineering, University of Science and Technology Beijing, Beijing 100083 (China); Zhao, Hailei [School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083 (China); Rafique, M. Yasir [Department of Physics, School of Applied Science, University of Science and Technology Beijing, Beijing 100083 (China); Rafi ud Din [School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083 (China); Chemistry Division, PINSTECH, P.O. Box Nilore, Islamabad (Pakistan); Farooq, M. Hassan [School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083 (China); Javed, Qurat ul ain [Department of Physics, School of Applied Science, University of Science and Technology Beijing, Beijing 100083 (China); Khan, Dil Faraz [School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083 (China)


    Graphical abstract: Display Omitted Highlights: ► Self-assembled stannous oxide (SnO) 2D nano-square sheets have been synthesized. ► The average size of the nano-square sheets is about 200–400 nm. ► Direct optical band gap of 3.16 eV was acquired by using Davis–Mott model. ► The hydrogen absorption capacity of as-synthesized material was 1.194 wt.% comparatively good. -- Abstract: Stannous oxide is an important functional material which contributes to a wide range of applications in energy storage and optoelectronic devices. In the present study, the single crystalline self-assembled stannous oxide (SnO) 2D nano-square sheets have been synthesized with template-free hydrothermal growth method. The morphology, composition and structure were characterized by field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), high resolution transmission electron microscopy (HRTEM) with selected area electron diffraction (SAED), energy dispersive X-ray (EDX), X-ray diffraction (XRD) and Raman spectroscopy, respectively. FESEM results have illustrated that the size of self-assembled 3D hierarchical polygon-shape structure of SnO is in the range of 8–12 μm and the average size of the nano-square sheets is about 100 nm. X-ray diffraction (XRD) and selected area electron diffraction (SAED) patterns have revealed that the prepared SnO nano-square sheets exist in single-crystalline nature. Two Raman modes A{sub 1g} = 211 cm{sup −1} and B{sub 1g} = 113 cm{sup −1} were observed by Raman spectroscopy, which is consistent with nano tetragonal phase SnO. Furthermore, the chemical valence of Sn and relative atomic composition of as-prepared SnO have been confirmed by X-ray photoelectron spectroscopy (XPS). Ultraviolet–visible–near infrared spectrophotometry was used to study the transmittance behavior of SnO nano-structures and direct optical band gap of 3.16 eV was acquired by using Davis–Mott model. The first ever study on

  10. Self-assembled nanostructures

    CERN Document Server

    Zhang, Jin Z; Liu, Jun; Chen, Shaowei; Liu, Gang-yu


    Nanostructures refer to materials that have relevant dimensions on the nanometer length scales and reside in the mesoscopic regime between isolated atoms and molecules in bulk matter. These materials have unique physical properties that are distinctly different from bulk materials. Self-Assembled Nanostructures provides systematic coverage of basic nanomaterials science including materials assembly and synthesis, characterization, and application. Suitable for both beginners and experts, it balances the chemistry aspects of nanomaterials with physical principles. It also highlights nanomaterial-based architectures including assembled or self-assembled systems. Filled with in-depth discussion of important applications of nano-architectures as well as potential applications ranging from physical to chemical and biological systems, Self-Assembled Nanostructures is the essential reference or text for scientists involved with nanostructures.

  11. H-Bonding Self-assembled Template-directed Synthesis of a Reactive Amide-bridged Ladder Polyvinylsiloxane

    Institute of Scientific and Technical Information of China (English)

    You Zhi WAN; Ying Hua LIU; Ping XIE; Rong Ben ZHANG


    A novel, reactive amide-bridged ladder polyvinylsiloxane (abbr. LP) with Mn = 2.4×104was synthesized for the first time by means of aryl amide H-bonding self-assembled template.The regularity of LP was characterized by the XRD, 29Si NMR and DSC methods. XRD analysis demonstrated the ladder width w = 9.09 A and the ladder thickness t = 3.89 A, respectively, which are approximately consistent with the molecular simulation-calculated ones: w'= 10.60 A and t'=3.06 A. 29Si NMR displayed a resonance peak with small half peak width, △1/2 ~ 4 ppm, for the moiety [=Si(Vi)O2/2-]n of LP. Besides, as a collateral evidence, DSC measurement revealed a high glass transition temperature Tg = 225℃, suggesting high stiffness of the ladder main chain of LP.

  12. Dansyl-labeled anionic amphiphile with a hexadecanoic carbon chain: synthesis and detection for shape transitions in organized molecular assemblies. (United States)

    Gao, Lining; Xia, Huiyun; Wang, Xiaoman; Li, Li; Chen, Huaxin


    The probing properties of a new fluorophore-labeled anionic surfactant, sodium 16-(N-dansyl)aminocetylate (16-DAN-ACA) were investigated systematically in molecular assemblies, especially in the transitions between micelles and vesicles. 16-DAN-ACA can efficiently differentiate the two different aggregate types in mixed cationic and anionic surfactant systems. The fluorescence anisotropy of 16-DAN-ACA was found to be sensitive for directly detecting the micellar growth in micelles containing oppositely charged surfactants; both cationic cetyltrimethylammonium bromide (CTAB) systems and anionic sodium dodecyl sulfate (SDS) systems were studied. The results indicated that the 16-DAN-ACA is a good fluorescent probe for differentiating the different aggregates, and even more can be used to detect the micellar growth.

  13. Miktoarms hyperbranched polymer brushes:One-step fast synthesis by parallel click chemistry and hierarchical self-assembly

    Institute of Scientific and Technical Information of China (English)


    Spherical molecular brushes with amphiphilic heteroarms were facilely synthesized by grafting the arms of hydrophobic 2-azidoethyle palmitate and hydrophilic monoazide-terminated poly(ethylene glycol) onto the core of alkyne-modified hyperbranched polyglycerol (HPG) with high molecular weight (Mn=122 kDa) via one-pot parallel click chemistry.The parallel click grafting strategy was demonstrated to be highly efficient (~100%),very fast (~ 2 h) and well controllable to the amphilicity of molecular brushes.Through adjusting the feeding ratio of hydrophobic and hydrophilic arms,a series of brushes with different arm ratios were readily obtained.The resulting miktoarms hyperbranched polymer brushes (HPG-g-C16/PEG350) were characterized by hydrogen-nuclear magnetic resonance (1H NMR),Fourier transform infrared (FT-IR) spectroscopy,gel permeation chromatography (GPC),and differential scanning calorimetry (DSC) measurements.The spherical molecular brushes showed high molecular weights up to 230 kDa,and thus could be visualized by atomic force microscopy (AFM).AFM and dynamic laser light scattering (DLS) were employed to investigate the self-assembly properties of amphiphilic molecular brushes with closed proportion of hydrophobic and hydrophilic arms.The brushes could self-assemble hierarchically into spherical micelles,and network-like fibre structures,and again spherical micelles by addition of n-hexane into the dichloromethane or chloroform solution of brushes.In addition,this kind of miktoarms polymer brush also showed the ability of dye loading via host-guest encapsulation,which promises the potential application of spherical molecular brushes in supramolecular chemistry.

  14. Cyclotetrabenzoin: Facile Synthesis of a Shape-Persistent Molecular Square and Its Assembly into Hydrogen-Bonded Nanotubes. (United States)

    Ji, Qing; Le, Ha T M; Wang, Xiqu; Chen, Yu-Sheng; Makarenko, Tatyana; Jacobson, Allan J; Miljanić, Ognjen Š


    Cyanide-catalyzed benzoin condensation of terephthaldehyde produces a cyclic tetramer, which we propose to name cyclotetrabenzoin. Cyclotetrabenzoin is a square-shaped macrocycle ornamented with four α-hydroxyketone functionalities pointing away from the central cavity, the dimensions of which are 6.9×6.9 Å. In the solid state, these functional groups extensively hydrogen bond, resulting in a microporous three-dimensional organic framework with one-dimensional nanotube channels. This material exhibits permanent-albeit low-porosity, with a Langmuir surface area of 52 m(2)  g(-1) . Cyclotetrabenzoin's easy and inexpensive synthesis and purification may inspire the creation of other shape-persistent macrocycles and porous molecular crystals by benzoin condensation.

  15. From Self-Assembled Monolayers to Coatings: Advances in the Synthesis and Nanobio Applications of Polymer Brushes

    Directory of Open Access Journals (Sweden)

    Myungwoong Kim


    Full Text Available In this review, we describe the latest advances in synthesis, characterization, and applications of polymer brushes. Synthetic advances towards well-defined polymer brushes, which meet criteria such as: (i Efficient and fast grafting, (ii Applicability on a wide range of substrates; and (iii Precise control of surface initiator concentration and hence, chain density are discussed. On the characterization end advances in methods for the determination of relevant physical parameters such as surface initiator concentration and grafting density are discussed. The impact of these advances specifically in emerging fields of nano- and bio-technology where interfacial properties such as surface energies are controlled to create nanopatterned polymer brushes and their implications in mediating with biological systems is discussed.

  16. Nitrite attenuated hypochlorous acid-mediated heme degradation in hemoglobin. (United States)

    Lu, Naihao; Li, Jiayu; Ren, Xiaoming; Tian, Rong; Peng, Yi-Yuan


    Hypochlorous acid (HOCl) is elevated in many inflammatory diseases and causes the accumulation of free iron. Through the Fenton reaction, free iron has the ability to generate free radicals and subsequently is toxic. Recent studies have demonstrated that HOCl participates in heme destruction of hemoglobin (Hb) and free iron release. In this study, it was showed that nitrite (NO2(-)) could prevent HOCl-mediated Hb heme destruction and free iron release. Also, NO2(-) prevented HOCl-mediated loss of Hb peroxidase activity. After the NO2(-)/HOCl treatment, Tyr 42 in α-chain was found to be nitrated in Hb, attenuating the electron transferring abilities of phenolic compounds. The protective effects of NO2(-) on HOCl-induced heme destruction were attributed to its reduction of ferryl Hb and/or direct scavenging of HOCl. Therefore, NO2(-) could show protective effects in some inflammatory diseases by preventing HOCl-mediated heme destruction of hemoproteins and free iron release.

  17. Immunolocalization of heme oxygenase-1 in periodontal diseases

    Directory of Open Access Journals (Sweden)

    G Gayathri


    Conclusion: The results of our study is an increasing evidence of involvement of antioxidant enzymes like heme oxygenase-1 in periodontal inflammation and their implication for treatment of chronic periodontitis.

  18. Cytochrome c peroxidase activity of heme bound amyloid β peptides. (United States)

    Seal, Manas; Ghosh, Chandradeep; Basu, Olivia; Dey, Somdatta Ghosh


    Heme bound amyloid β (Aβ) peptides, which have been associated with Alzheimer's disease (AD), can catalytically oxidize ferrocytochrome c (Cyt c(II)) in the presence of hydrogen peroxide (H2O2). The rate of catalytic oxidation of Cyt(II) c has been found to be dependent on several factors, such as concentration of heme(III)-Aβ, Cyt(II) c, H2O2, pH, ionic strength of the solution, and peptide chain length of Aβ. The above features resemble the naturally occurring enzyme cytochrome c peroxidase (CCP) which is known to catalytically oxidize Cyt(II) c in the presence of H2O2. In the absence of heme(III)-Aβ, the oxidation of Cyt(II) c is not catalytic. Thus, heme-Aβ complex behaves as CCP.

  19. Mechanisms of heme iron absorption: Current questions and controversies

    Institute of Scientific and Technical Information of China (English)


    Iron is a critical micronutrient, and iron derived from heme contributes a large proportion of the total iron absorbed in a typical Western diet. Heine iron is absorbed by different mechanisms than non-heine iron, but despite considerable study over many years these mechanisms remain poorly understood. This review provides an overview of the importance of heme iron in the diet and discusses the two prevailing hypotheses of heine absorption; namely receptor mediated endocytosis of heme, and direct transport into the intestinal enterocyte by recently discovered heine transporters. A specific emphasis is placed on the questions surrounding the site of heme catabolism and the identity of the enzyme that performs this task. Additionally, we present the hypothesis that a nonheme iron transport protein may be required for heine iron absorption and discuss the experiences of our laboratory in examining this hypothesis.

  20. Structure of the Escherichia coli O157:H7 heme oxygenase ChuS in complex with heme and enzymatic inactivation by mutation of the heme coordinating residue His-193

    Energy Technology Data Exchange (ETDEWEB)

    Suits,M.; Jaffer, N.; Jia, Z.


    Heme oxygenases catalyze the oxidation of heme to biliverdin, CO, and free iron. For pathogenic microorganisms, heme uptake and degradation are critical mechanisms for iron acquisition that enable multiplication and survival within hosts they invade. Here we report the first crystal structure of the pathogenic Escherichia coli O157:H7 heme oxygenase ChuS in complex with heme at 1.45 {angstrom} resolution. When compared with other heme oxygenases, ChuS has a unique fold, including structural repeats and a {beta}-sheet core. Not surprisingly, the mode of heme coordination by ChuS is also distinct, whereby heme is largely stabilized by residues from the C-terminal domain, assisted by a distant arginine from the N-terminal domain. Upon heme binding, there is no large conformational change beyond the fine tuning of a key histidine (His-193) residue. Most intriguingly, in contrast to other heme oxygenases, the propionic side chains of heme are orientated toward the protein core, exposing the {alpha}-meso carbon position where O{sub 2} is added during heme degradation. This unique orientation may facilitate presentation to an electron donor, explaining the significantly reduced concentration of ascorbic acid needed for the reaction. Based on the ChuS-heme structure, we converted the histidine residue responsible for axial coordination of the heme group to an asparagine residue (H193N), as well as converting a second histidine to an alanine residue (H73A) for comparison purposes. We employed spectral analysis and CO measurement by gas chromatography to analyze catalysis by ChuS, H193N, and H73A, demonstrating that His-193 is the key residue for the heme-degrading activity of ChuS.

  1. Genome-wide analysis reveals novel genes essential for heme homeostasis in Caenorhabditis elegans.

    Directory of Open Access Journals (Sweden)

    Scott Severance


    Full Text Available Heme is a cofactor in proteins that function in almost all sub-cellular compartments and in many diverse biological processes. Heme is produced by a conserved biosynthetic pathway that is highly regulated to prevent the accumulation of heme--a cytotoxic, hydrophobic tetrapyrrole. Caenorhabditis elegans and related parasitic nematodes do not synthesize heme, but instead require environmental heme to grow and develop. Heme homeostasis in these auxotrophs is, therefore, regulated in accordance with available dietary heme. We have capitalized on this auxotrophy in C. elegans to study gene expression changes associated with precisely controlled dietary heme concentrations. RNA was isolated from cultures containing 4, 20, or 500 microM heme; derived cDNA probes were hybridized to Affymetrix C. elegans expression arrays. We identified 288 heme-responsive genes (hrgs that were differentially expressed under these conditions. Of these genes, 42% had putative homologs in humans, while genomes of medically relevant heme auxotrophs revealed homologs for 12% in both Trypanosoma and Leishmania and 24% in parasitic nematodes. Depletion of each of the 288 hrgs by RNA-mediated interference (RNAi in a transgenic heme-sensor worm strain identified six genes that regulated heme homeostasis. In addition, seven membrane-spanning transporters involved in heme uptake were identified by RNAi knockdown studies using a toxic heme analog. Comparison of genes that were positive in both of the RNAi screens resulted in the identification of three genes in common that were vital for organismal heme homeostasis in C. elegans. Collectively, our results provide a catalog of genes that are essential for metazoan heme homeostasis and demonstrate the power of C. elegans as a genetic animal model to dissect the regulatory circuits which mediate heme trafficking in both vertebrate hosts and their parasites, which depend on environmental heme for survival.

  2. Design, synthesis, and properties of a new class of building blocks for assembly of molecule-based magnets (United States)

    Misiolek, Andrzej Wladyslaw

    In this work, mono and dianions of the tetrone 1 were proposed as discrete building blocks for assembly of molecular magnets.* These anionic subunits are designed to form extended multidimensional structures upon complexation with metal rations. This particular framework also was chosen so that dianion 12- (or its derivatives) would possess a paramagnetic triplet as a ground state. High quality ab-initio CASSCF(14, 12)/6-31+G* calculations showed that indeed the singlet-triplet gap of metal complexes of 1 2- is small and its sign and magnitude tongly depends on structural modifications. For example a 17.3 kcal/mol preference for a singlet is predicted for isolated 12- while the triplet state is preferred by 4.3 kcal/mol for the symmetrically coordinated complex 1(AlF2)2.* Synthetic procedures leading to a family of neutral aryl substituted tetrones (2) has been developed and electrochemical studies have shown that they may be reversibly reduced in two successive one-electron processes to mono- and dianions. These mono- and dianions have been generated on a preparative scale by reduction of the parent tetrones with alkali metals under anaerobic conditions. Solutions of anions were characterized by Vis-NIR, NMR and EPR spectroscopy. EPR of the anion radicals 2- reveals the high symmetry of the spin delocalisation. Spectroscopic studies of dianions were hampered by low solubility and aggregation phenomena and no definite answer concerning their ground spin states could be obtained. Several attempts were made to grow single crystals of 2 - and 22- in order to probe the self-assembling properties of those species and to obtain definite answers concerning the ground state of the dianion. These attempts turned out to be successful only for alkali metal salts of the monoanion (t-Bu) 8(MeO)42-. Its K + salt crystallized as 1-D chains with the expected bridging and chelating mode of coordination. The closely related Na+ salt, recently crystallized by Robert Gentner, shows a

  3. HmuS and HmuQ of Ensifer/Sinorhizobium meliloti degrade heme in vitro and participate in heme metabolism in vivo. (United States)

    Amarelle, Vanesa; Rosconi, Federico; Lázaro-Martínez, Juan Manuel; Buldain, Graciela; Noya, Francisco; O'Brian, Mark R; Fabiano, Elena


    Ensifer meliloti is a nitrogen-fixing symbiont of the alfalfa legume able to use heme as an iron source. The transport mechanism involved in heme acquisition in E. meliloti has been identified and characterized, but the fate of heme once inside the cell is not known. In silico analysis of E. meliloti 1021 genome revealed no canonical heme oxygenases although two genes encoding putative heme degrading enzymes, smc01518 and hmuS, were identified. SMc01518 is similar to HmuQ of Bradyrhizobium japonicum, which is weakly homologous to the Staphylococcus aureus IsdG heme-degrading monooxygenase, whereas HmuS is homolog to Pseudomonas aeruginosa PhuS, a protein reported as a heme chaperone and as a heme degrading enzyme. Recombinant HmuQ and HmuS were able to bind hemin with a 1:1 stoichiometry and displayed a Kd value of 5 and 4 µM, respectively. HmuS degrades heme in vitro to the biliverdin isomers IX-β and IX-δ in an equimolar ratio. The HmuQ recombinant protein degrades heme to biliverdin IX-δ only. Additionally, in this work we demonstrate that humS and hmuQ gene expression is regulated by iron and heme in a RirA dependent manner and that both proteins are involved in heme metabolism in E. meliloti in vivo.

  4. On-Surface Synthesis of Two-Dimensional Covalent Organic Structures versus Halogen-Bonded Self-Assembly: Competing Formation of Organic Nanoarchitectures. (United States)

    Peyrot, David; Silly, Fabien


    The competition between the on-surface synthesis of covalent nanoarchitectures and the self-assembly of star-shaped 1,3,5-Tris(4-iodophenyl)benzene molecules on Au(111) in vacuum is investigated using scanning tunneling microscopy above room temperature. The molecules form covalent polygonal nanoachitectures at the gold surface step edges and at the elbows of the gold reconstruction at low coverage. With coverage increasing two-dimensional halogen-bonded structures appear and grow on the surface terraces. Two different halogen-bonded nanoarchitectures are coexisting on the surface and hybrid covalent-halogen bonded structures are locally observed. At high coverage covalent nanoarchitectures are squeezed at the domain boundary of the halogen-bonded structures. The competitive growth between the covalent and halogen-bonded nanoarchitectures leads to formation of a two-layer film above one monolayer deposition. For this coverage, the covalent nanoarchitectures are propelled on top of the halogen-bonded first layer. These observations open up new opportunities for decoupling covalent nanoarchitectures from catalytically active and metal surfaces in vacuum.

  5. Rapid and One-Pot Synthesis of Self-Assembled CdSe Quantum Dots Functionalized with β-Cyclodextrin: Reduced Cytotoxicity and Band Gap Engineering. (United States)

    Guleria, Apurav; Rath, Madhab C; Singh, Ajay K; Adhikari, Soumyakanti


    We report a simple, rapid and one step method for the synthesis and in situ functionalization of CdSe quantum dots (QDs) with β-cyclodextrin (β-CD) in aqueous solution via electron beam (EB) irradiation technique. A probable mechanism has been elucidated for the formation of the QDs using pulse radiolysis technique. The average size of the QDs was found to be in the range of 2-3 nm with a size distribution of -14%. XPS measurements indicate that the -OH groups of the β-CD molecules binds predominantly with the Cd atoms present on the surface of the QDs. These QDs displayed broad photoluminescence (PL) with two emission peaks at 525 nm and 600 nm, which could be tuned by varying the experimental parameters. The broad PL spectrum has been attributed to the polydispersity in the density and the distribution of trap/defects states. Time resolved PL decay measurements further substantiated the domination of surface state originated carrier relaxation processes in the overall PL decay dynamics of QDs synthesized at higher doses and dose rates. The present study reveals that β-CD passivate the QDs by a non-inclusion complex, induces the self-assembling process into a networking architecture and simultaneously reduces their cytotoxicity as compared to the bare nanoparticles. The methodology described in this article may provide unique and interesting aspects to regulate and fine tune the formation of superstructures of nanomaterials vis-à-vis their optoelectronic properties.

  6. Miktoarm star copolymers from D-(-)-salicin core aggregated into dandelion-like structures as anticancer drug delivery systems: synthesis, self-assembly and drug release. (United States)

    Mielańczyk, Anna; Odrobińska, Justyna; Grządka, Sebastian; Mielańczyk, Łukasz; Neugebauer, Dorota


    The β-glucoside-based heterofunctional initiator was used in the synthesis of well-defined eight-armed miktopolymers by sequential ring opening polymerization (ROP) of ε-caprolactone (CL) and atom transfer radical (co)polymerization (ATRP) of methyl methacrylate (MMA) and/or tert-butyl methacrylate (tBMA). Consequently, methacrylic acid (MAA) repeating units were introduced via selective cleavage of pendant tert-butyl protecting groups. Both the amphiphilic copolymers and miktoarm copolymers were self-assembled at 37°C and pH 7.4. The aggregates of miktoarm polymers were larger than that formed by polymethacrylate homoarm stars (≥250nm vs ≤200nm). The critical aggregation concentrations (CAC) of (mikto)stars were relatively low (0.006-0.411mg/mL) and decreased with the increase in MAA fraction content. Both MAA-based mikto- and homoarmed (co)polymers with shorter arms exhibited lower doxorubicin (DOX) loading capacity, whereas camptothecin (CPT) was encapsulated preferably by miktostars. The kinetic profiles of drug release showed that the rate of release was higher at acidic environment (pH 5.0) than in neutral pH. In the most cases the studied miktopolymer systems demonstrated the well-controlled delivery of the model anticancer drugs, which can be adjusted by structural parameters of polymeric carriers.

  7. Decreased Heme Oxygenase Activity in Patients with Alzheimer's Disease

    Directory of Open Access Journals (Sweden)

    Berkay Cataloglu


    Full Text Available Alzheimer's disease is a neurodegenerative disorder characterized with progressive im-pairment of cognitive functions. Heme oxygenase is an enzyme that degrades the heme molecule resulting in equimolar amounts of the carbon monoxide, ferrous iron, and bili-verdin. Up to now, heme oxygenase activity and its metabolic effects in Alzheimer's dis-ease have been investigated in so many studies; most of them were performed in post-mortem brain tissues of Alzheimer's disease patients or in animal models. Therefore, we aimed to investigate heme oxygenase activity in leukocytes of Alzheimer's disease pa-tients as a peripheral sample. Mean heme oxygenase activity was significantly lower in patients with Alzheimer's disease (0.53 +/- 0.32 nmol/h/mg protein compared to control sucjects (1.19 +/- 0.84 nmol/h/mg protein (p= 0.001. We think that reduction in leukocyte heme oxygenase activity may limit disease progression through preserving peripheral mitochondrial function by reducing the formation of free iron and carbon monoxide. [Dis Mol Med 2013; 1(2.000: 31-34

  8. Alterations in renal heme biosynthesis during metal nephrotoxicity. (United States)

    Oskarsson, A; Fowler, B A


    The regulation of the heme biosynthetic pathway in the kidney by various metals has been reviewed. In addition, a study on the effects of lead on renal heme biosynthesis after acute treatment of rats has been reported. Chronic low-level lead exposure in rats results in relatively small effects on renal heme biosynthetic pathway enzymes. After acute treatment of rats with lead, no effects on ALAD or UROS and mild, transitory effects on ALAS and ferrochelatase are observed. The intracellular binding of lead within intranuclear inclusion bodies in the proximal tubule cells and to high-affinity cytosolic lead-binding proteins probably protects sensitive subcellular systems, such as the heme pathway, from lead toxicity. Chronic exposure to methyl mercury results in increased urinary excretion of uro- and coproporphyrins in rats, mediated via inhibition of ferrochelatase and UROS and stimulation of ALAS. A tissue-specific inhibition of ALAD occurs in the kidney after treatment of rats with indium. Acute treatment of rats with nickel, platinum, tin, antimony, bismuth, and cobalt results in induction of heme oxygenase, followed by decreased microsomal heme content and ALAS stimulation in the kidney.

  9. Scalable synthesis of hierarchical macropore-rich activated carbon microspheres assembled by carbon nanoparticles for high rate performance supercapacitors (United States)

    Zhang, Dongdong; Zhao, Jianghong; Feng, Chong; Zhao, Rijie; Sun, Yahui; Guan, Taotao; Han, Baixin; Tang, Nan; Wang, Jianlong; Li, Kaixi; Qiao, Jinli; Zhang, Jiujun


    A scalable inverse-microemulsion-polymerization-phase-separation coupling method is applied to successfully prepare hierarchical macropore-rich activated carbon microspheres (ACS) using a phenolic resin (PR) precursor followed by carbonization and KOH activation for the first time. The formed ACS materials are assembled by carbon nanoparticles (CNPs). The macropores interspersed among the component CNPs are formed after removing the non-reactive solvent phase in the course of the polymerization of the reactive PR phase, which occupies ∼64% of the total pore volume (∼2.779 cm3 g-1) of the optimized ACS. In combination with mesopores (∼18% of the total pore volume), the ACS possesses meso/macropores approaching 82% of the total pore volume. Micropores are created in the component CNPs via KOH activation, showing shortened ion transport distances in the nanoscale dimension. Both the hierarchical micro/meso/macroporous structure and the inner nanoparticle morphology (short ion diffusion pathways) can significantly contribute to the rapid transport of electrolyte ions throughout the carbonaceous matrix, resulting in superior rate performance of ACS-based supercapacitors. More importantly, the energy densities of the ACS supercapacitors operating in both aqueous and organic electrolyte retain steady over a wide range of power densities varying dramatically from 0.25 to 14.5 kW kg-1 and to 7.0 kW kg-1, respectively.

  10. Synthesis and characterization of mesoporous spinel NiCo2O4 using surfactant-assembled dispersion for asymmetric supercapacitors (United States)

    Hsu, Chun-Tsung; Hu, Chi-Chang


    A simple and scalable process has been developed for synthesizing spinel NiCo2O4 nanocrystals through a thermal decomposition method. The introduction of hexadecyltrimethylammonium bromide (CTAB, (C16H33)N(CH3)3Br) into precursor solutions significantly enhances the homogeneity and porosity of spinel NiCo2O4. The porosity and high specific surface area of NiCo2O4 preserves the brilliant pseudo-capacitive performances due to providing smooth paths for electrolyte penetration and ion diffusion into inner active sites. Morphologies and microstructures of the active materials are examined by transmission electron microscopic (TEM) and X-ray diffraction (XRD) analyses. Thermogravimetric analysis (TGA) is used to evaluate the thermal properties of precursor solutions. The electrochemical performances of NiCo2O4 are systematically characterized by cyclic voltammetry and charge-discharge tests. Asymmetric supercapacitors are assembled with these brilliant binary oxides as the positive electrode and activated carbon as the negative electrode. The highly porous NiCo2O4 exhibits superior capacitive performances, i.e., high specific capacitance (764 F g-1 at 2 mV s-1) and long cycle life.

  11. Synthesis of visible light emitting self assembled Ge nanocrystals embedded within a SiO{sub 2} matrix

    Energy Technology Data Exchange (ETDEWEB)

    Hernandez-Hernandez, A.; De Moure-Flores, F.; Quinones-Galvan, J. G.; Santoyo-Salazar, J.; Melendez-Lira, M. [Departamento de Fisica, Centro de Investigacion y de Estudios Avanzados del Instituto Politecnico Nacional, A.P. 14740, C.P. 07300, Mexico, Distrito Federal (Mexico); Rangel-Kuoppa, V. T. [Institute of Semiconductor and Solid State Physics, Johannes Kepler Universitaet, A-4040 Linz (Austria); Plach, Thomas [Christian Doppler Laboratory for Microscopic and Spectroscopic Material Characterization, Center for Surface and Nanoanalytics, Johannes Kepler Universitaet, A-4040 Linz (Austria); Zapata-Torres, M. [Centro de Investigacion en Ciencia Aplicada y Tecnologia Avanzada, Unidad Legaria IPN, Calzada Legaria 694, Col. Irrigacion, 11500 Mexico, Distrito Federal (Mexico); Hernandez-Hernandez, L. A. [Escuela Superior de Fisica y Matematicas del Instituto Politecnico Nacional, Edificio 9 U.P. Adolfo Lopez Mateos, Col. San Pedro Zacatenco, C.P. 07730 (Mexico)


    As-grown light emitting self-assembled Ge nanocrystals (Ge-NCs) embedded in a SiO{sub 2} matrix were produced via a sequential deposition process of SiO{sub 2}/Ge/SiO{sub 2} layers employing a reactive radio frequency sputtering technique. Obtained Ge-NCs show a crystallographic phase, the proportion, size, quality, and specific orientation of which are determined by the oxygen partial pressure. Photoluminescence (PL) spectra indicate that the size distribution of Ge-NCs is reduced and centered on about 8 nm when higher oxygen partial pressure is employed; the formation of Ge-NCs is corroborated by transmission electron microscopy measurements, and their sizes are consistent with estimates from PL measurements. Resistivity measurements are explained by a near neighbors hopping process, with specific features depending on the Ge-NCs' size. The features of PL and resistivity measurements indicate that there is no appreciable dependence of the number of interfacial defects on the oxygen partial pressure.

  12. Synthesis of visible light emitting self assembled Ge nanocrystals embedded within a SiO2 matrix (United States)

    Hernández-Hernández, A.; Rangel-Kuoppa, V. T.; Plach, Thomas; De Moure-Flores, F.; Quiñones-Galván, J. G.; Santoyo-Salazar, J.; Zapata-Torres, M.; Hernández-Hernández, L. A.; Meléndez-Lira, M.


    As-grown light emitting self-assembled Ge nanocrystals (Ge-NCs) embedded in a SiO2 matrix were produced via a sequential deposition process of SiO2/Ge/SiO2 layers employing a reactive radio frequency sputtering technique. Obtained Ge-NCs show a crystallographic phase, the proportion, size, quality, and specific orientation of which are determined by the oxygen partial pressure. Photoluminescence (PL) spectra indicate that the size distribution of Ge-NCs is reduced and centered on about 8 nm when higher oxygen partial pressure is employed; the formation of Ge-NCs is corroborated by transmission electron microscopy measurements, and their sizes are consistent with estimates from PL measurements. Resistivity measurements are explained by a near neighbors hopping process, with specific features depending on the Ge-NCs' size. The features of PL and resistivity measurements indicate that there is no appreciable dependence of the number of interfacial defects on the oxygen partial pressure.


    Energy Technology Data Exchange (ETDEWEB)

    Ramirez-Ruiz, Enrico; MacLeod, Morgan [Department of Astronomy and Astrophysics, University of California, Santa Cruz, CA 95064 (United States); Trenti, Michele [Kavli Institute for Cosmology and Institute of Astronomy, University of Cambridge, Madingley Road, Cambridge CB3 0HA (United Kingdom); Roberts, Luke F. [TAPIR, California Institute of Technology, Pasadena, California 91125 (United States); Lee, William H.; Saladino-Rosas, Martha I. [Instituto de Astronomía, Universidad Nacional Autónoma de México, México DF 04510, México (Mexico)


    Investigations of elemental abundances in the ancient and most metal deficient stars are extremely important because they serve as tests of variable nucleosynthesis pathways and can provide critical inferences of the type of stars that lived and died before them. The presence of r-process elements in a handful of carbon-enhanced metal-poor (CEMP-r) stars, which are assumed to be closely connected to the chemical yield from the first stars, is hard to reconcile with standard neutron star mergers. Here we show that the production rate of dynamically assembled compact binaries in high-z nuclear star clusters can attain a sufficient high value to be a potential viable source of heavy r-process material in CEMP-r stars. The predicted frequency of such events in the early Galaxy, much lower than the frequency of Type II supernovae but with significantly higher mass ejected per event, can naturally lead to a high level of scatter of Eu as observed in CEMP-r stars.

  14. Synthesis of Ionic Liquid Based Electrolytes, Assembly of Li-ion Batteries, and Measurements of Performance at High Temperature. (United States)

    Lin, Xinrong; Chapman Varela, Jennifer; Grinstaff, Mark W


    The chemical instability of the traditional electrolyte remains a safety issue in widely used energy storage devices such as Li-ion batteries. Li-ion batteries for use in devices operating at elevated temperatures require thermally stable and non-flammable electrolytes. Ionic liquids (ILs), which are non-flammable, non-volatile, thermally stable molten salts, are an ideal replacement for flammable and low boiling point organic solvent electrolytes currently used today. We herein describe the procedures to: 1) synthesize mono- and di-phosphonium ionic liquids paired with chloride or bis(trifluoromethane)sulfonimide (TFSI) anions; 2) measure the thermal properties and stability of these ionic liquids by differential scanning calorimetry (DSC) and thermal gravimetric analysis (TGA); 3) measure the electrochemical properties of the ionic liquids by cyclic voltammetry (CV); 4) prepare electrolytes containing lithium bis(trifluoromethane)sulfonamide; 5) measure the conductivity of the electrolytes as a function of temperature; 6) assemble a coin cell battery with two of the electrolytes along with a Li metal anode and LiCoO2 cathode; and 7) evaluate battery performance at 100 °C. We additionally describe the challenges in execution as well as the insights gained from performing these experiments.

  15. Synthesis of porphyrinoids with silane anchors and their covalent self-assembling and metallation on solid surface. (United States)

    Sariola-Leikas, Essi; Hietala, Matti; Veselov, Alexey; Okhotnikov, Oleg; Semjonov, Sergei L; Tkachenko, Nikolai V; Lemmetyinen, Helge; Efimov, Alexander


    We have synthesized a set of porphyrin and phthalocyanine compounds with two different silane anchors. Syntheses of the anchor-substituted chromophores have been carried out via hydrosilylation of alkene derivatives, catalyzed by platinum complexes. The reduction side-process was suppressed using specific anchor/catalyst pairs, and the silane-containing compounds were successfully isolated from hydrogenated by-products in pure form with good yields. The target porphyrinoids having stable reactive silane anchors possess the ability to self-assemble on metal oxides and quartz surfaces and optical fibers. Covalent attachment is done in one-step, which makes the bonding process fast and easy. Immobilized chromophores were further converted by on-surface reactions into Zn(II) and Mg(II) metal complexes. The metallation time was found to be as fast as 1 min for Zn ion. Bonding densities calculated from the absorbances of the deposited layers give rough estimations for packing of the molecules on various substrates and evidence for monomolecular layers formation.

  16. Synthesis of Inorganic Nanocomposites by Selective Introduction of Metal Complexes into a Self-Assembled Block Copolymer Template

    Directory of Open Access Journals (Sweden)

    Hiroaki Wakayama


    Full Text Available Inorganic nanocomposites have characteristic structures that feature expanded interfaces, quantum effects, and resistance to crack propagation. These structures are promising for the improvement of many materials including thermoelectric materials, photocatalysts, and structural materials. Precise control of the inorganic nanocomposites’ morphology, size, and chemical composition is very important for these applications. Here, we present a novel fabrication method to control the structures of inorganic nanocomposites by means of a self-assembled block copolymer template. Different metal complexes were selectively introduced into specific polymer blocks of the block copolymer, and subsequent removal of the block copolymer template by oxygen plasma treatment produced hexagonally packed porous structures. In contrast, calcination removal of the block copolymer template yielded nanocomposites consisting of metallic spheres in a matrix of a metal oxide. These results demonstrate that different nanostructures can be created by selective use of processes to remove the block copolymer templates. The simple process of first mixing block copolymers and magnetic nanomaterial precursors and then subsequently removing the block copolymer template enables structural control of magnetic nanomaterials, which will facilitate their applicability in patterned media, including next-generation perpendicular magnetic recording media.

  17. Self-assembly and template-free synthesis of ZnO hierarchical nanostructures and their photocatalytic properties. (United States)

    Zhou, Hongshun; Zhang, Haijiao; Wang, Yong; Miao, Yu; Gu, Lanbing; Jiao, Zheng


    Despite significant progress in the field of semiconductor photocatalysis has been made, it is still a great challenge to prepare low-cost photocatalysts with high activities. In our work, three dimensional (3D) flower-like ZnO hierarchical nanostructures assembled with numerous nanosheets were fabricated by a simple, template-free and one-step hydrothermal route. The products were characterized by XRD, UV-Vis, PL, SEM, TEM, HRTEM techniques. In the process, NH4F played a crucial role for the formation of ZnO hierarchical nanostructures, which was acted both as the alkali source and morphology director. Furthermore, the growth of ZnO involved a phase transformation from intermediate compound ZnF(OH) to ZnO. To further improve the photocatalytic activity, Ag-doped ZnO photocatalyst was also prepared. The photocatalytic results indicated that the Ag/ZnO exhibited higher photocatalytic activity than the pure ZnO. The great enhancement was mainly ascribed to their unique hierarchical nanostructures as well as the modification of Ag nanoparticles. Additionally, both ZnO and Ag/ZnO microspheres showed good recycling stabilities over several separation cycles in photodegradation.

  18. Synthesis and self-assembly of spin-labile and redox-active manganese(III) complexes. (United States)

    Gandolfi, Claudio; Cotting, Tatiana; Martinho, Paulo N; Sereda, Olha; Neels, Antonia; Morgan, Grace G; Albrecht, Martin


    New amphiphilic and spin-labile Mn(III) complexes based on dianionic N(4)O(2)-hexadentate sal(2)trien or sal(2)bapen ligands, which contain OC(6)H(13), OC(12)H(25), or OC(18)H(37) alkoxy substituents at different positions of the salicylidene unit were prepared (H(2)sal(2)trien = N,N'''-bis(salicylidene)-1,4,7,10-tetraazadecane, H(2)sal(2)bapen = N,N'''-bis(salicylidene)-1,5,8,12-tetraazadodecane). According to electrochemical measurements, these complexes undergo two (quasi)reversible redox processes. Temperature-dependent magnetic measurements revealed a high-spin configuration for all sal(2)trien complexes (S = 2) and gradual spin crossover for sal(2)bapen complexes from high to low spin (S = 1). The chain length strongly influences the spin crossover, as C(18)-functionalization stabilizes the low spin state at much higher temperatures than shorter alkyl chains. Moreover, long alkyl chains allow for spontaneous self-assembly of the molecules, which was investigated in single crystals and in Langmuir-films at the air-water interface. Long alkyl chains (C(12) or C(18)) as well as a mutual syn-orientation of these molecular recognition sites were required for the Langmuir monolayers to be stable.

  19. AtMYB41 activates ectopic suberin synthesis and assembly in multiple plant species and cell types. (United States)

    Kosma, Dylan K; Murmu, Jhadeswar; Razeq, Fakhria M; Santos, Patricia; Bourgault, Richard; Molina, Isabel; Rowland, Owen


    Suberin is a lipid and phenolic cell wall heteropolymer found in the roots and other organs of all vascular plants. Suberin plays a critical role in plant water relations and in protecting plants from biotic and abiotic stresses. Here we describe a transcription factor, AtMYB41 (At4g28110), that can activate the steps necessary for aliphatic suberin synthesis and deposition of cell wall-associated suberin-like lamellae in both Arabidopsis thaliana and Nicotiana benthamiana. Overexpression of AtMYB41 increased the abundance of suberin biosynthetic gene transcripts by orders of magnitude and resulted in the accumulation of up to 22 times more suberin-type than cutin-type aliphatic monomers in leaves. Overexpression of AtMYB41 also resulted in elevated amounts of monolignols in leaves and an increase in the accumulation of phenylpropanoid and lignin biosynthetic gene transcripts. Surprisingly, ultrastructural data indicated that overexpression led to the formation of suberin-like lamellae in both epidermal and mesophyll cells of leaves. We further implicate AtMYB41 in the production of aliphatic suberin under abiotic stress conditions. These results provide insight into the molecular-genetic mechanisms of the biosynthesis and deposition of a ubiquitous cell wall-associated plant structure and will serve as a basis for discovering the transcriptional network behind one of the most abundant lipid-based polymers in nature.

  20. The effect of proteins from animal source foods on heme iron bioavailability in humans. (United States)

    Pizarro, Fernando; Olivares, Manuel; Valenzuela, Carolina; Brito, Alex; Weinborn, Valerie; Flores, Sebastián; Arredondo, Miguel


    Forty-five women (35-45 year) were randomly assigned to three iron (Fe) absorption sub-studies, which measured the effects of dietary animal proteins on the absorption of heme Fe. Study 1 was focused on heme, red blood cell concentrate (RBCC), hemoglobin (Hb), RBCC+beef meat; study 2 on heme, heme+fish, chicken, and beef; and study 3 on heme and heme+purified animal protein (casein, collagen, albumin). Study 1: the bioavailability of heme Fe from Hb was similar to heme only (∼13.0%). RBCC (25.0%) and RBCC+beef (21.3%) were found to be increased 2- and 1.6-fold, respectively, when compared with heme alone (pheme alone (10.3%) was reduced (pheme Fe absorption.

  1. A heme-binding domain controls regulation of ATP-dependent potassium channels. (United States)

    Burton, Mark J; Kapetanaki, Sofia M; Chernova, Tatyana; Jamieson, Andrew G; Dorlet, Pierre; Santolini, Jérôme; Moody, Peter C E; Mitcheson, John S; Davies, Noel W; Schmid, Ralf; Raven, Emma L; Storey, Nina M


    Heme iron has many and varied roles in biology. Most commonly it binds as a prosthetic group to proteins, and it has been widely supposed and amply demonstrated that subtle variations in the protein structure around the heme, including the heme ligands, are used to control the reactivity of the metal ion. However, the role of heme in biology now appears to also include a regulatory responsibility in the cell; this includes regulation of ion channel function. In this work, we show that cardiac KATP channels are regulated by heme. We identify a cytoplasmic heme-binding CXXHX16H motif on the sulphonylurea receptor subunit of the channel, and mutagenesis together with quantitative and spectroscopic analyses of heme-binding and single channel experiments identified Cys628 and His648 as important for heme binding. We discuss the wider implications of these findings and we use the information to present hypotheses for mechanisms of heme-dependent regulation across other ion channels.

  2. Altered activity of heme biosynthesis pathway enzymes in individuals chronically exposed to arsenic in Mexico

    Energy Technology Data Exchange (ETDEWEB)

    Hernandez-Zavala, A.; Del Razo, L.M.; Garcia-Vargas, G.G.; Aguilar, C.; Borja, V.H.; Albores, A.; Cebrian, M.E. [CINVESTAV-IPN, Mexico (Mexico). Dept. de Farmacologia y Toxicologica


    Our objective was to evaluate the activities of some enzymes of the heme biosynthesis pathway and their relationship with the profile of urinary porphyrin excretion in individuals exposed chronically to arsenic (As) via drinking water in Region Lagunera, Mexico. We selected 17 individuals from each village studied: Benito Juarez, which has current exposure to 0.3 mg As/l; Santa Ana, where individuals have been exposed for more than 35 years to 0.4 mg As/l, but due to changes in the water supply (in 1992) exposure was reduced to its current level (0.1 mg As/l), and Nazareno, with 0.014 mg As/l. Average arsenic concentrations in urine were 2058, 398, and 88 {mu}g As/g creatinine, respectively. The more evident alterations in heme metabolism observed in the highly exposed individuals were: (1) small but significant increases in porphobilinogen deaminase (PBG-D) and uroporphyrinogen decarboxylase (URO-D) activities in peripheral blood erythrocytes; (2) increases in the urinary excretion of total porphyrins, mainly due to coproporphyrin III (COPROIII) and uroporphyrin III (UROIII); and (3) increases in the COPRO/URO and COPROIII/COPROI ratios. No significant changes were observed in uroporphyrinogen III synthetase (UROIII-S) activity. The direct relationships between enzyme activities and urinary porphyrins, suggest that the increased porphyrin excretion was related to PBG-D, whereas the increased URO-D activity would enhance coproporphyrin synthesis and excretion at the expense of uroporphyrin. None of the human studies available have reported the marked porphyric response and enzyme inhibition observed in rodents. In conclusion, chronic As exposure alters human heme metabolism; however the severity of the effects appears to depend on characteristics of exposure not yet fully characterized. (orig.) With 1 fig., 3 tabs., 20 refs.

  3. Altered activity of heme biosynthesis pathway enzymes in individuals chronically exposed to arsenic in Mexico. (United States)

    Hernández-Zavala, A; Del Razo, L M; García-Vargas, G G; Aguilar, C; Borja, V H; Albores, A; Cebrián, M E


    Our objective was to evaluate the activities of some enzymes of the heme biosynthesis pathway and their relationship with the profile of urinary porphyrin excretion in individuals exposed chronically to arsenic (As) via drinking water in Region Lagunera, Mexico. We selected 17 individuals from each village studied: Benito Juarez, which has current exposure to 0.3 mg As/l; Santa Ana, where individuals have been exposed for more than 35 years to 0.4 mg As/l, but due to changes in the water supply (in 1992) exposure was reduced to its current level (0.1 mg As/l), and Nazareno, with 0.014 mg As/l. Average arsenic concentrations in urine were 2058, 398, and 88 microg As/g creatinine, respectively. The more evident alterations in heme metabolism observed in the highly exposed individuals were: (1) small but significant increases in porphobilinogen deaminase (PBG-D) and uroporphyrinogen decarboxylase (URO-D) activities in peripheral blood erythrocytes; (2) increases in the urinary excretion of total porphyrins, mainly due to coproporphyrin III (COPROIII) and uroporphyrin III (UROIII); and (3) increases in the COPRO/URO and COPROIII/COPROI ratios. No significant changes were observed in uroporphyrinogen III synthetase (UROIII-S) activity. The direct relationships between enzyme activities and urinary porphyrins, suggest that the increased porphyrin excretion was related to PBG-D, whereas the increased URO-D activity would enhance coproporphyrin synthesis and excretion at the expense of uroporphyrin. None of the human studies available have reported the marked porphyric response and enzyme inhibition observed in rodents. In conclusion, chronic As exposure alters human heme metabolism; however the severity of the effects appears to depend on characteristics of exposure not yet fully characterized.

  4. Analysis of Heme oxygenase isomers in rat

    Institute of Scientific and Technical Information of China (English)

    Zhen-Wei Xia; Wen-Jun Cui; Xue-Hong Zhang; Qing-Xiang Shen; Jian Wang; Yun-Zhu Li; Shen-Nian Chen; Shan-Chang Yu


    AIM: To purify and identify heme oxygenase (HO) isomers which exist in rat liver, spleen and brain treated with hematin and phenylhydrazine and in untreated rat liver and to investigate the characteristics of HO isomers, to isolate and confirm the rat HO-1 cDNA that actually encodes HO-1 by expressing cDNA in monkey kidney cells (COS-1 cells), to prepare the rat heme oxygenase-1 (HO-1) mutant and to detect inhibition of HO-1 mutated enzyme.METHODS: First, rat liver, spleen and brain microsomal fi-actions were purified by DEAE-Sephacel and hydroxylapatite. The characteristics including activity, immunity and inducibility of two isomers (HO-1 and HO-2), and their apparent molecular weight were measured by detecting enzymatic activities, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting analysis, respectively. Second, plasmid pcDNA3HO1 containing native rat HO-1 cDNA and pcDNA3HO1D25 carrying mutated rat HO-1 cDNA (His25Ala) were constructed by site-directed mutagenesis. COS-1 cells transfected with pcDNA3HO1 and pcDNA3HO1D25 were collected and disrupted by sonication, the microsomes were prepared by ultracentrifugation. Third, the inhibition of rat HO-1 mutant was analyzed.RESULTS: Two isomers were purified and identified in treated rat liver, spleen, brain and untreated rat liver. HO-1 was the predominant form with a ratio of 2.0:1 and 3.2:1 of HO-1 and HO-2 in liver and spleen, respectively, but only the activity of HO-2 in the brain and untreated liver could be detected. The apparent molecular weights of HO-1 and HO-2 were about Mr 30 000 and Mr 36 000 under reducing conditions, respectively. The antiserum against liver HO-2 was employed in Western blotting analysis, the reactivity of HO-1 in the liver was not observed. The plasmid pcDNA3HO1 was highly expressed in endoplasmic reticulum of transfected COS-1 cells. The specific activity was ≈5-fold higher than that of the control. However, the enzyme activity of mutated HO-1 declined. While

  5. Synthesis, biocompatible, and self-assembly properties of poly (ethylene glycol)/lactobionic acid-grafted chitosan. (United States)

    Song, Xiaoli; Wang, Juan; Luo, Xiadan; Xu, Chunlan; Zhu, Aiping; Guo, Rong; Yan, Caifeng; Zhu, Peizhi


    Polymers with targeted ligands are widely used as the anti-cancer drug delivery materials. For applications of chitosan as an anti-liver cancer drug delivery, poly (ethylene glycol)/lactobionic acid-grafted chitosan (PEG/LA-CS) was prepared and investigated since lactobionic acid can be specifically recognized by the hepatocytes. The structure of the PEG/LA-CS was characterized by Fourier transform infrared spectrometry and elemental analysis. The self-assembly behaviors of the PEG/LA-CS were monitored by steady-state fluorescence spectroscopy and electronic transmission microscope. The protein adsorption of the PEG/LA-CS was detected with bovine serum albumin (BSA) by electrochemical impedance spectroscopy. The results showed that the PEG/LA-CS almost did not adsorb protein. To study the effects of PEG/LA-CS on the structure of BSA, the interactions between the PEG/LA-CS and BSA were detected by ultraviolet spectrum, fluorescence spectrum, and circular dichroism. All the data gave one result that BSA maintained its original folded confirmation in PEG/LA-CS solution. The hemocompatibility of PEG/LA-CS was investigated by observing the effects of PEG/LA-CS on the hemolysis rate and the plasma recalcification time (PRT). The results showed that the PRT was prolonged greatly and the hemolysis rate was less than 5%. Furthermore, PEG/LA-CS also showed good cytocompatibility with K562, Hep G2, and LO2 cells. Therefore, the PEG/LA-CS is believed to have great potential for producing injectable anti-liver cancer drug delivery.

  6. Self-assembly synthesis,crystal structure and nonlinear optical properties of cluster compound containing PPh2Py ligand

    Institute of Scientific and Technical Information of China (English)

    ZHOU Jian-liang; WEI Chuan-jun; TONG Hai-xia; CHUN Xiao-gai; CHEN Qi-yuan


    Self-assembly cluster compound [WS4Cu3(PPh2Py)3Br]2·CH3OH (1) was synthesized by the reaction of (NH4)2WS4,CuBr and diphenyl-2-pyridyl-phosphine (PPh2Py) in CH3OH solution under a purified nitrogen atmosphere using standard Schlenk techniques.Its structure was determined by X-ray crystallography.It crystallizes in the triclinic crystal system P-1 space group with α=1.178 6(1)nm,b=1.302 6(1)nm,c=1.9917(2)nm,α=74.671(7)°,β=86.188(8)°,γ=64.141(6)°,V=2.649 5(5)nm',Z=1.The W center is slightly distorted from tetrahedral coordination geometry,and the structure is built up from three [Cu(PPh2Py)]+ units bridged by WS2-4 multifunctional ligand to form a tetranuclear symmetrical cube-like molecule.Measurement of the nonlinear optical (NLO) properties using the Z-scan technique with an 8 ns pulsed laser at 532 nm shows that the compound possesses NLO absorption and effective self-focusing effect at α2=6.7×10-11m/W and n2=5.64×10-18m2/W in a 1.5×10-4mol/L DMF solution.

  7. Synthesis and Crystal Structure of a New Double 2D Layer Complex Assembled by H4btec

    Institute of Scientific and Technical Information of China (English)


    Assembly of H4btec with cobalt(Ⅱ) acetate afforded a new double 2D layer coor- dination polymer, [Co(btec)1/2(H2O)4·2H2O]n (H4btec = benzene-1,2,4,5-tetracarboxylic acid) 1. The polymeric structure has been characterized by single-crystal X-ray diffraction, FT-IR and thermal analysis. The crystal is of triclinic, space group P with a = 10.8762(2), b = 11.1411(1), c = 11.5084(3)(A), α = 82.8950(10), β = 63.0050(10), γ = 62.1500(10)°, V = 1091.72(4)(A)3, C5H13CoO10, Mr = 292.08, Z = 4, Dc = 1.777 g/cm3, μ = 1.612 mm-1, F(000) = 600, R = 0.0578, and wR = 0.2162 for 3576 observed reflections (I > 2σ(I)). Complex 1 consists of a new double 2D layer structure [Co(btec)1/2(H2O)4]n, both consisting of 4 + 4 grids with the sizes of 11.3667(2)(A) × 11.5084(3)(A)(Co(1)-Co(1A) × Co(2A)-Co(2B)) and 11.5084(3)(A)× 11.3667(2)(A) (Co(3)-Co(3C) × Co(4)- Co(4A)), respectively. The phenyl rings are at the corners while the Co(Ⅱ) atoms are in the sides of the grids, and lattice water molecules decorate between the layers. Hydrogen bonds between the layers and lattice water molecules result in the final 3D framework.

  8. Heme Degradation by Heme Oxygenase Protects Mitochondria but Induces ER Stress via Formed Bilirubin

    Directory of Open Access Journals (Sweden)

    Andrea Müllebner


    Full Text Available Heme oxygenase (HO, in conjunction with biliverdin reductase, degrades heme to carbon monoxide, ferrous iron and bilirubin (BR; the latter is a potent antioxidant. The induced isoform HO-1 has evoked intense research interest, especially because it manifests anti-inflammatory and anti-apoptotic effects relieving acute cell stress. The mechanisms by which HO mediates the described effects are not completely clear. However, the degradation of heme, a strong pro-oxidant, and the generation of BR are considered to play key roles. The aim of this study was to determine the effects of BR on vital functions of hepatocytes focusing on mitochondria and the endoplasmic reticulum (ER. The affinity of BR to proteins is a known challenge for its exact quantification. We consider two major consequences of this affinity, namely possible analytical errors in the determination of HO activity, and biological effects of BR due to direct interaction with protein function. In order to overcome analytical bias we applied a polynomial correction accounting for the loss of BR due to its adsorption to proteins. To identify potential intracellular targets of BR we used an in vitro approach involving hepatocytes and isolated mitochondria. After verification that the hepatocytes possess HO activity at a similar level as liver tissue by using our improved post-extraction spectroscopic assay, we elucidated the effects of increased HO activity and the formed BR on mitochondrial function and the ER stress response. Our data show that BR may compromise cellular metabolism and proliferation via induction of ER stress. ER and mitochondria respond differently to elevated levels of BR and HO-activity. Mitochondria are susceptible to hemin, but active HO protects them against hemin-induced toxicity. BR at slightly elevated levels induces a stress response at the ER, resulting in a decreased proliferative and metabolic activity of hepatocytes. However, the proteins that are targeted

  9. 新型超分子化合物的合成自组装及应用研究的新进展%Recent Research Achievements on Synthesis,Self-assembly and Applications of New Supramolecular Compounds

    Institute of Scientific and Technical Information of China (English)

    张来新; 胡小兵


    This paper briefly introduced the definition, concept, generation and application of supramolecular chemistry. Emphases were put on three parts:① synthesis and self-assembly of new supramolecular compounds;② synthesis and selective recognition effects of new supramolecular compounds;③synthesis and application of su-pramolecular crown ether metal complexes.%简要介绍了超分子化学的定义、概念、产生及应用,详细介绍了:①新型超分子化合物合成及自组装;②新型超分子化合物的合成及选择性识别作用;③超分子冠醚金属配合物的合成及应用。

  10. Heme-binding plasma membrane proteins of K562 erythroleukemia cells: Adsorption to heme-microbeads, isolation with affinity chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Majuri, R. (Minerva Foundation Institute for Medical Research, Helsinki (Finland))


    Heme-microbeads attached themselves to the surface of viable K562 cells in a manner inhibitable by free hemin, indicating heme-recptor interaction. The microbeads were at first evenly distributed, but after prolonged incubation at 37 deg. C they formed a cap on one pole of the cells indicating clustering of the membrane heme receptors. Membrane proteins were labeled by culturing the cells in the presence of {sup 35}S-methionine and were then solubilized with Triton X-114. The hydrophobic proteins contained about 20% of the total bound label. The solubilized membrane proteins were subsequently adsorbed to a heme-Sepharose affinity gel. According to SDS-electrophorsis and subsequent autoradiography, the immobilized heme captures two proteins or a protein with two polypeptides of 20 000 and 32 000 daltons. The larger of these was only wekly labeled with {sup 35}S. The same two bands were observed if the cell surface proteins were labeled with {sup 125}I by the lactoperoxidase method and the subsequently solubilized membrane proteins were isolated with heme-Sepharose. (author).

  11. The Role of the Cytoplasmic Heme-binding Protein (PhuS) of Pseudomonas aeruginosa in Intracellular Heme Trafficking and Iron Homeostasis*S⃞



    The cytoplasmic heme-binding protein PhuS, encoded within the Fur-regulated Pseudomonas heme utilization (phu) operon, has previously been shown to traffic heme to the iron-regulated heme oxygenase (HO). We further investigate the role of PhuS in heme trafficking to HO on disruption of the phuS and hemO genes in a Pseudomonas aeruginosa siderophore-deficient and wild-type background. Previous studies have shown that deletion of hemO prevents the cells from utilizin...

  12. Homologues of Neisserial Heme Oxygenase in Gram-Negative Bacteria: Degradation of Heme by the Product of the pigA Gene of Pseudomonas aeruginosa



    The oxidative cleavage of heme to release iron is a mechanism by which some bacterial pathogens can utilize heme as an iron source. The pigA gene of Pseudomonas aeruginosa is shown to encode a heme oxygenase protein, which was identified in the genome sequence by its significant homology (37%) with HemO of Neisseria meningitidis. When the gene encoding the neisserial heme oxygenase, hemO, was replaced with pigA, we demonstrated that pigA could functionally replace hemO and allow for heme util...

  13. Synthesis, Characterization and Application of Poly (Styrene-4- Vinyl Pyridine) Membranes Assembled With Single-Wall Carbon Nanotubes

    KAUST Repository

    He, Haoze


    Poly(styrene‐4‐vinylpyridine) (PS‐P4VP) isoporous membranes were prepared and their properties were evaluated in this research. The solution was prepared by dissolving PS‐P4VP polymer with necessary additives into a 1:1:1 1,4‐dioxane – N,N‐dimethyl formamide – tetrahydrofuran (DOX‐DMF‐THF, DDT) solvent. Then 0.5‐1.0 mL of the primary solution was cast onto the non‐woven substrate membrane on a glass slide, evaporated for 15‐20 sec and immersed into de‐ionized water for more than 30 min for the solidification of isoporous structure and for the formation of the primary films, which could be post‐processed in different ways for different tests. The membrane surface presents a well‐ordered, hexagonal self‐assembly structure, which is fit for aqueous and gaseous filtration. The pore size of the isoporous surface is 30~40 nm. The pore size is also sensitive to [H+] in the solution and a typical pair of S‐shape pH‐correlation curves with significant hysteresis was found. Four techniques were tried to improve the properties of the membranes in this research: 1) 1,4‐diiodobutane was introduced to chemically change the structure as a cross‐linking agent. 2) single‐wall carbon nanotube (SWCNT) was linked to the membranes in order to strengthen the stability and rigidity and to reduce the hysteresis. 3) Homo‐poly(4‐vinylpyridine) (homo‐P4VP) was added and inserted into the PS‐P4VP micelles to affect the pore size and surface structure. 4) Copper acetate (Cu(Ac)2) was used as substitute of dioxane to prepare the Cu(Ac)2‐DMF‐THF (CDT) mixed solvent, for a better SWCNT dispersion. All the possible improvements were judged by the atomic force microscopy (AFM) images, water and gas flux tests and pH‐correlation curves. The introduction of SWCNT was the most important innovation in this research and is promising in future applications.

  14. Doxorubicin-loaded aromatic imine-contained amphiphilic branched star polymer micelles: synthesis, self-assembly, and drug delivery

    Directory of Open Access Journals (Sweden)

    Qiu L


    Full Text Available Liang Qiu, Chun-Yan Hong, Cai-Yuan Pan Chinese Academy of Sciences Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei, Anhui, People’s Republic of China Abstract: Redox- and pH-sensitive branched star polymers (BSPs, BP(DMAEMA-co-MAEBA-co-DTDMA(PMAIGPns, have been successively prepared by two steps of reversible addition–fragmentation chain transfer (RAFT polymerization. The first step is RAFT polymerization of 2-(N,N-dimethylaminoethylmethacrylate (DMAEMA and p-(methacryloxyethoxybenzaldehyde (MAEBA in the presence of divinyl monomer, 2,2'-dithiodiethoxyl dimethacrylate (DTDMA. The resultant branched polymers were used as a macro-RAFT agent in the subsequent RAFT polymerization. After hydrolysis of the BSPs to form BP(DMAEMA-co-MAEBA-co-DTDMA(PMAGPns (BSP-H, the anticancer drug doxorubicin (DOX was covalently linked to branched polymer chains by reaction of primary amine of DOX and aldehyde groups in the polymer chains. Their compositions, structures, molecular weights, and molecular weight distributions were respectively characterized by nuclear magnetic resonance spectra and gel permeation chromatography measurements. The DOX-loaded micelles were fabricated by self-assembly of DOX-containing BSPs in water, which were characterized by transmission electron microscopy and dynamic light scattering. Aromatic imine linkage is stable in neutral water, but is acid-labile; controlled release of DOX from the BSP-H-DOX micelles was realized at pH values of 5 and 6, and at higher acidic solution, fast release of DOX was observed. In vitro cytotoxicity experiment results revealed low cytotoxicity of the BSPs and release of DOX from micelles in HepG2 and HeLa cells. Confocal laser fluorescence microscopy observations showed that DOX-loaded micelles have specific interaction with HepG2 cells. Thus, this type of BSP micelle is an efficient drug delivery system

  15. α-Hemoglobin-stabilizing Protein (AHSP) Perturbs the Proximal Heme Pocket of Oxy-α-hemoglobin and Weakens the Iron-Oxygen Bond* (United States)

    Dickson, Claire F.; Rich, Anne M.; D'Avigdor, William M. H.; Collins, Daniel A. T.; Lowry, Jason A.; Mollan, Todd L.; Khandros, Eugene; Olson, John S.; Weiss, Mitchell J.; Mackay, Joel P.; Lay, Peter A.; Gell, David A.


    α-Hemoglobin (αHb)-stabilizing protein (AHSP) is a molecular chaperone that assists hemoglobin assembly. AHSP induces changes in αHb heme coordination, but how these changes are facilitated by interactions at the αHb·AHSP interface is not well understood. To address this question we have used NMR, x-ray absorption spectroscopy, and ligand binding measurements to probe αHb conformational changes induced by AHSP binding. NMR chemical shift analyses of free CO-αHb and CO-αHb·AHSP indicated that the seven helical elements of the native αHb structure are retained and that the heme Fe(II) remains coordinated to the proximal His-87 side chain. However, chemical shift differences revealed alterations of the F, G, and H helices and the heme pocket of CO-αHb bound to AHSP. Comparisons of iron-ligand geometry using extended x-ray absorption fine structure spectroscopy showed that AHSP binding induces a small 0.03 Å lengthening of the Fe-O2 bond, explaining previous reports that AHSP decreases αHb O2 affinity roughly 4-fold and promotes autooxidation due primarily to a 3–4-fold increase in the rate of O2 dissociation. Pro-30 mutations diminished NMR chemical shift changes in the proximal heme pocket, restored normal O2 dissociation rate and equilibrium constants, and reduced O2-αHb autooxidation rates. Thus, the contacts mediated by Pro-30 in wild-type AHSP promote αHb autooxidation by introducing strain into the proximal heme pocket. As a chaperone, AHSP facilitates rapid assembly of αHb into Hb when βHb is abundant but diverts αHb to a redox resistant holding state when βHb is limiting. PMID:23696640

  16. α-Hemoglobin-stabilizing protein (AHSP) perturbs the proximal heme pocket of oxy-α-hemoglobin and weakens the iron-oxygen bond. (United States)

    Dickson, Claire F; Rich, Anne M; D'Avigdor, William M H; Collins, Daniel A T; Lowry, Jason A; Mollan, Todd L; Khandros, Eugene; Olson, John S; Weiss, Mitchell J; Mackay, Joel P; Lay, Peter A; Gell, David A


    α-Hemoglobin (αHb)-stabilizing protein (AHSP) is a molecular chaperone that assists hemoglobin assembly. AHSP induces changes in αHb heme coordination, but how these changes are facilitated by interactions at the αHb·AHSP interface is not well understood. To address this question we have used NMR, x-ray absorption spectroscopy, and ligand binding measurements to probe αHb conformational changes induced by AHSP binding. NMR chemical shift analyses of free CO-αHb and CO-αHb·AHSP indicated that the seven helical elements of the native αHb structure are retained and that the heme Fe(II) remains coordinated to the proximal His-87 side chain. However, chemical shift differences revealed alterations of the F, G, and H helices and the heme pocket of CO-αHb bound to AHSP. Comparisons of iron-ligand geometry using extended x-ray absorption fine structure spectroscopy showed that AHSP binding induces a small 0.03 Å lengthening of the Fe-O2 bond, explaining previous reports that AHSP decreases αHb O2 affinity roughly 4-fold and promotes autooxidation due primarily to a 3-4-fold increase in the rate of O2 dissociation. Pro-30 mutations diminished NMR chemical shift changes in the proximal heme pocket, restored normal O2 dissociation rate and equilibrium constants, and reduced O2-αHb autooxidation rates. Thus, the contacts mediated by Pro-30 in wild-type AHSP promote αHb autooxidation by introducing strain into the proximal heme pocket. As a chaperone, AHSP facilitates rapid assembly of αHb into Hb when βHb is abundant but diverts αHb to a redox resistant holding state when βHb is limiting.

  17. Facile heme vinyl posttranslational modification in a hemoglobin. (United States)

    Preimesberger, Matthew R; Wenke, Belinda B; Gilevicius, Lukas; Pond, Matthew P; Lecomte, Juliette T J


    Iron-protoporphyrin IX, or b heme, is utilized as such by a large number of proteins and enzymes. In some cases, notably the c-type cytochromes, this group undergoes a posttranslational covalent attachment to the polypeptide chain, which adjusts the physicochemical properties of the holoprotein. The hemoglobin from the cyanobacterium Synechocystis sp. PCC 6803 (GlbN), contrary to the archetypical hemoglobin, modifies its b heme covalently. The posttranslational modification links His117, a residue that does not coordinate the iron, to the porphyrin 2-vinyl substituent and forms a hybrid b/c heme. The reaction is an electrophilic addition that occurs spontaneously in the ferrous state of the protein. This apparently facile type of heme modification has been observed in only two cyanobacterial GlbNs. To explore the determinants of the reaction, we examined the behavior of Synechocystis GlbN variants containing a histidine at position 79, which is buried against the porphyrin 4-vinyl substituent. We found that L79H/H117A GlbN bound the heme weakly but nevertheless formed a cross-link between His79 Nε2 and the heme 4-Cα. In addition to this linkage, the single variant L79H GlbN also formed the native His117-2-Cα bond yielding an unprecedented bis-alkylated protein adduct. The ability to engineer the doubly modified protein indicates that the histidine-heme modification in GlbN is robust and could be engineered in different local environments. The rarity of the histidine linkage in natural proteins, despite the ease of reaction, is proposed to stem from multiple sources of negative selection.

  18. Ultrafast Spectroscopy Evidence for Picosecond Ligand Exchange at the Binding Site of a Heme Protein: Heme-Based Sensor YddV. (United States)

    Lambry, Jean-Christophe; Stranava, Martin; Lobato, Laura; Martinkova, Marketa; Shimizu, Toru; Liebl, Ursula; Vos, Marten H


    An important question for the functioning of heme proteins is whether different ligands present within the protein moiety can readily exchange with heme-bound ligands. Studying the dynamics of the heme domain of the Escherichia coli sensor protein YddV upon dissociation of NO from the ferric heme by ultrafast spectroscopy, we demonstrate that when the hydrophobic leucine residue in the distal heme pocket is mutated to glycine, in a substantial fraction of the protein water replaces NO as an internal ligand in as fast as ∼4 ps. This process, which is near-barrierless and occurs orders of magnitude faster than the corresponding process in myoglobin, corresponds to a ligand swap of NO with a water molecule present in the heme pocket, as corroborated by molecular dynamics simulations. Our findings provide important new insight into ligand exchange in heme proteins that functionally interact with different external ligands.

  19. Anion-controlled assembly of metal 3,5-bis(benzimidazol-1-ylmethyl) benzoate complexes: Synthesis, characterization and property

    Energy Technology Data Exchange (ETDEWEB)

    Kuai, Hai-Wei [Coordination Chemistry Institute, State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering, Nanjing National Laboratory of Microstructures, Nanjing University, Nanjing 210093 (China); Faculty of Life Science and Chemical Engineering, Huaiyin Institute of Technology, Huaian 223003 (China); Lv, Gao-Chao; Hou, Chao [Coordination Chemistry Institute, State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering, Nanjing National Laboratory of Microstructures, Nanjing University, Nanjing 210093 (China); Sun, Wei-Yin, E-mail: [Coordination Chemistry Institute, State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering, Nanjing National Laboratory of Microstructures, Nanjing University, Nanjing 210093 (China)


    Hydrothermal reactions of 3,5-bis(benzimidazol-1-ylmethyl)benzoic acid (HL) with Cd(II), Cu(II) and Zn(II) salts provide eight new metal complexes which were characterized by single crystal and powder X-ray diffraction, IR, elemental and thermogravimetric analyses. Two cadmium frameworks [Cd(L){sub 2}]·2H{sub 2}O (1) and [Cd(L)Cl] (2) have 3D structures with (4{sup 2}.6{sup 5}.8{sup 3})(4{sup 2}.6) and rtl (4.6{sup 2}){sub 2}(4{sup 2}.6{sup 10}.8{sup 3}) topologies, respectively. Structural diversity of four copper complexes [Cu{sub 3}(L){sub 2}]·NO{sub 3}·0.5H{sub 2}O (3), [Cu{sub 2}(HL){sub 2}(SO{sub 4})]·3.5H{sub 2}O (4), [Cu(L)(bdc){sub 0.5}]·1.5H{sub 2}O (5) and [Cu{sub 2}(L)(HL)(Hbdc)] (6) (H{sub 2}bdc=1,4-benzenedicarboxylic acid) is achieved through the alteration of copper salts and addition of auxiliary ligand. As a result, 3 has a 1D ladder structure, 4 is a discrete dinuclear complex, 5 displays a (3,4)-connected 2-nodal 3-fold interpenetrating framework with (4{sup 2}.6.10{sup 2}.12)(4{sup 2}.6) topology, 6 exhibits a 4-connected uninodal 2D sql (4{sup 4}.6{sup 2}) network. Within the zinc series, ZnCl{sub 2} and ZnSO{sub 4} were used for the syntheses of [Zn(L)Cl] (7) and [Zn(L)(SO{sub 4}){sub 0.5}]·2H{sub 2}O (8), respectively. 7 shows a 3-connected uninodal 2D hcb network with (6{sup 3}) topology and 8 is a (3,6)-connected 2-nodal 3D framework with (4{sup 2}.6){sub 2}(4{sup 4}.6{sup 2}.8{sup 8}.10) topology. The luminescent properties of the Cd(II) and Zn(II) complexes were investigated. - Graphical abstract: Eight new complexes have been successfully synthesized from the hydrothermal reactions of Cd(II), Cu(II) and Zn(II) salts with 3,5-bis(benzimidazol-1-ylmethyl)benzoic acid. The complexes exhibited anion-controlled structural diversity. - Highlights: • Metal complexes have diverse structures of 1D chains, 2D networks and 3D frameworks. • Anion-controlled assembly of the complexes is reported. • The luminescent properties of the Cd

  20. KH2PO4-Assisted Synthesis and Electrochemical Performance of Highly Uniform CuBi2O4 Microspheres Hierarchically Self-Assembled by Nanoparticles (United States)

    Wang, Fei; Yang, Hua; Zhang, Yunchuan; Zhang, Haimin


    The effect of KH2PO4 on the synthesis of CuBi2O4 microstructures was investigated. The samples were characterized by powder x-ray diffraction (XRD), scanning electron microscopy (SEM), Brunauer-Emmett-Teller (BET) analysis, x-ray photoelectron spectroscopy (XPS) and ultraviolet (UV)-visible diffuse reflectance spectroscopy. It is demonstrated that the use of KH2PO4 leads to the production of highly uniform CuBi2O4 microspheres hierarchically self-assembled by nanoparticles. With increasing the KH2PO4 concentration from 0.5 M to 1.4 M, the average diameter of the resultant microspheres decreases gradually from 3.3 μm to 1.4 μm. However, further increase in the KH2PO4 concentration up to 1.5 M leads to a sudden increase in the average diameter of the resultant microspheres up to 2.3 μm. In addition, a minor amount of bamboo leaf- or pine needle-like structures are visible in the samples prepared at the KH2PO4 concentrations of 1.0-1.5 M. The bandgap energy of the as-prepared samples is measured to be 1.89 eV by UV-visible diffuse reflectance spectroscopy. The electrochemical performance of the samples was investigated by cyclic voltammetry, galvanostatic charge-discharge measurements, and electrochemical impedance spectroscopy in 2 M KOH electrolyte. Among the hierarchical microspheres, those prepared at the KH2PO4 concentration of 1.4 M deliver a relatively higher specific capacitance due to their smaller size (1284 F g-1 at a current density of 2 A g-1).

  1. Synthesis and Optical Properties of Assembly-Controlled ZnO Structures%ZnO微结构调控及其光学性能的研究

    Institute of Scientific and Technical Information of China (English)

    刘海霞; 黄柏标; 王泽岩; 秦晓燕; 张晓阳; 俞娇仙


    A facile solvothermal synthesis of ZnO architectures has been developed using ethanol as solvent and tartaric acid as the additive. The variational quantities of tartaric acid controlled the ZnO morphology and shape as well as the self-assembly of ZnO crystals into complex architectures. Possible growth mechanisms of obtained ZnO with different morphologies were proposed. The influence of tartaric acid to ZnO growth was further indicated by FTIR spectra. In addition, the photoluminescence (PL) properties of these ZnO samples were investigated at room temperature, which indicated that the ZnO morphology could influence its optical property. On the whole, the fluorescence was violet emission and orange emission primarily.%氧化锌不同形貌的合成与控制可以通过一个简单的溶剂热反应来实现,其中乙醇作溶剂,酒石酸做添加剂.通过控制酒石酸的加入量,可以有效地控制ZnO的形貌、尺寸以及到更复杂结构的转变.同时提出了不同形貌ZnO可能的生长机制,并利用FFIR谱进一步证实了酒石酸对ZnO生长的影响.另外,由光致发光光谱可以看出,不同的ZnO形貌,发光性能会有所不同,总体上说,所得ZnO的发光区域主要集中在紫光波段和橙光波段.

  2. Heme oxygenase-1 deletion affects stress erythropoiesis.

    Directory of Open Access Journals (Sweden)

    Yu-An Cao

    Full Text Available BACKGROUND: Homeostatic erythropoiesis leads to the formation of mature red blood cells under non-stress conditions, and the production of new erythrocytes occurs as the need arises. In response to environmental stimuli, such as bone marrow transplantation, myelosuppression, or anemia, erythroid progenitors proliferate rapidly in a process referred to as stress erythropoiesis. We have previously demonstrated that heme oxygenase-1 (HO-1 deficiency leads to disrupted stress hematopoiesis. Here, we describe the specific effects of HO-1 deficiency on stress erythropoiesis. METHODOLOGY/PRINCIPAL FINDINGS: We used a transplant model to induce stress conditions. In irradiated recipients that received hmox(+/- or hmox(+/+ bone marrow cells, we evaluated (i the erythrocyte parameters in the peripheral blood; (ii the staining intensity of CD71-, Ter119-, and CD49d-specific surface markers during erythroblast differentiation; (iii the patterns of histological iron staining; and (iv the number of Mac-1(+-cells expressing TNF-α. In the spleens of mice that received hmox(+/- cells, we show (i decreases in the proerythroblast, basophilic, and polychromatophilic erythroblast populations; (ii increases in the insoluble iron levels and decreases in the soluble iron levels; (iii increased numbers of Mac-1(+-cells expressing TNF-α; and (iv decreased levels of CD49d expression in the basophilic and polychromatophilic erythroblast populations. CONCLUSIONS/SIGNIFICANCE: As reflected by effects on secreted and cell surface proteins, HO-1 deletion likely affects stress erythropoiesis through the retention of erythroblasts in the erythroblastic islands of the spleen. Thus, HO-1 may serve as a therapeutic target for controlling erythropoiesis, and the dysregulation of HO-1 may be a predisposing condition for hematologic diseases.

  3. Porphyrin-Cored Polymer Nanoparticles: Macromolecular Models for Heme Iron Coordination. (United States)

    Rodriguez, Kyle J; Hanlon, Ashley M; Lyon, Christopher K; Cole, Justin P; Tuten, Bryan T; Tooley, Christian A; Berda, Erik B; Pazicni, Samuel


    Porphyrin-cored polymer nanoparticles (PCPNs) were synthesized and characterized to investigate their utility as heme protein models. Created using collapsible heme-centered star polymers containing photodimerizable anthracene units, these systems afford model heme cofactors buried within hydrophobic, macromolecular environments. Spectroscopic interrogations demonstrate that PCPNs display redox and ligand-binding reactivity similar to that of native systems and thus are potential candidates for modeling biological heme iron coordination.

  4. Effect of Growth Conditions on Yield and Heme Content of Vitreoscilla


    Lamba, Parveen; Webster, Dale A.


    Vitreoscilla, a gliding bacterium in the Beggiatoaceae, is an obligate aerobe in which cytochrome o functions as the terminal oxidase. Protoheme IX is the only heme type present in this organism. The yield and heme content of Vitreoscilla cells grown in yeast extract, peptone, and acetate were dependent on growth conditions. Cells harvested in early stationary phase contained roughly three times as much heme as cells in early log phase. There was an optimal shaking rate for maximum heme conte...

  5. Heme-Scavenging Role of alpha1-Microglobulin in Chronic Ulcers.


    Allhorn, Maria; Lundqvist, Katarina; Schmidtchen, Artur; Åkerström, Bo


    Chronic venous ulcers are characterized by chronic inflammation. Heme and iron, originating from blood cell hemolysis as well as extravascular necrosis, have been implicated as important pathogenic factors due to their promotion of oxidative stress. It was recently reported that the plasma and tissue protein alpha1-microglobulin is involved in heme metabolism. The protein binds heme, and a carboxy-terminally processed form, truncated alpha1-microglobulin, also degrades heme. Here, we show the...

  6. Dysfunction of the heme recycling system in heme oxygenase 1-deficient mice: effects on macrophage viability and tissue iron distribution. (United States)

    Kovtunovych, Gennadiy; Eckhaus, Michael A; Ghosh, Manik C; Ollivierre-Wilson, Hayden; Rouault, Tracey A


    To better understand the tissue iron overload and anemia previously reported in a human patient and mice that lack heme oxygenase-1 (HO-1), we studied iron distribution and pathology in HO-1(Hmox1)(-/-) mice. We found that resident splenic and liver macrophages were mostly absent in HO-1(-/-) mice. Erythrophagocytosis caused the death of HO-1(-/-) macrophages in in vitro experiments, supporting the hypothesis that HO-1(-/-) macrophages died of exposure to heme released on erythrophagocytosis. Rupture of HO-1(-/-) macrophages in vivo and release of nonmetabolized heme probably caused tissue inflammation. In the spleen, initial splenic enlargement progressed to red pulp fibrosis, atrophy, and functional hyposplenism in older mice, recapitulating the asplenia of an HO-1-deficient patient. We postulate that the failure of tissue macrophages to remove senescent erythrocytes led to intravascular hemolysis and increased expression of the heme and hemoglobin scavenger proteins, hemopexin and haptoglobin. Lack of macrophages expressing the haptoglobin receptor, CD163, diminished the ability of haptoglobin to neutralize circulating hemoglobin, and iron overload occurred in kidney proximal tubules, which were able to catabolize heme with HO-2. Thus, in HO-1(-/-) mammals, the reduced function and viability of erythrophagocytosing macrophages are the main causes of tissue damage and iron redistribution.

  7. Degradation of heme in gram-negative bacteria: the product of the hemO gene of Neisseriae is a heme oxygenase. (United States)

    Zhu, W; Wilks, A; Stojiljkovic, I


    A full-length heme oxygenase gene from the gram-negative pathogen Neisseria meningitidis was cloned and expressed in Escherichia coli. Expression of the enzyme yielded soluble catalytically active protein and caused accumulation of biliverdin within the E. coli cells. The purified HemO forms a 1:1 complex with heme and has a heme protein spectrum similar to that previously reported for the purified heme oxygenase (HmuO) from the gram-positive pathogen Corynebacterium diphtheriae and for eukaryotic heme oxygenases. The overall sequence identity between HemO and these heme oxygenases is, however, low. In the presence of ascorbate or the human NADPH cytochrome P450 reductase system, the heme-HemO complex is converted to ferric-biliverdin IXalpha and carbon monoxide as the final products. Homologs of the hemO gene were identified and characterized in six commensal Neisseria isolates, Neisseria lactamica, Neisseria subflava, Neisseria flava, Neisseria polysacchareae, Neisseria kochii, and Neisseria cinerea. All HemO orthologs shared between 95 and 98% identity in amino acid sequences with functionally important residues being completely conserved. This is the first heme oxygenase identified in a gram-negative pathogen. The identification of HemO as a heme oxygenase provides further evidence that oxidative cleavage of the heme is the mechanism by which some bacteria acquire iron for further use.

  8. The heme oxygenase system and its functions in the brain. (United States)

    Maines, M D


    The heme oxygenase (HO) system was identified in the early 1970s as a distinct microsomal enzyme system that catalyzes formation of bile pigments (Maines and Kappas, 1974). Up to the early 1990s the system was considered only as a "molecular wrecking ball" (Lane, 1998) for degradation of the heme molecule and production of toxic waste products, CO and bile pigments. For those years, the HO system remained relatively unknown to the research community. In a rather short span of the past 10 years following the discovery of high levels of a second form of the enzyme, HO-2, in the brain, suggesting that "heme oxygenase in the brain has functions aside from heme degradation" (Sun et al., 1990); concomitant with finding that another toxic gas, NO, is a signal molecule for generation of cGMP (Ignarro et al., 1982), the system was propelled into main stream research. This propulsion was fueled by the realization of the multiple and diverse functions of heme degradation products. Heme oxygenase has now found relevance in all kinds of human pathophysiology ranging from stroke, cancer, multiple sclerosis, and malaria to transplantation and immune response. As it turns out, its potential benefits are mesmerizing investigators in diverse fields (Lane, 1998). The most recent findings with HO-2 being a hemoprotein and potentially an intracellular "sink" for NO (McCoubrey et al., 1997a; Ding et al., 1999), together with the discovery of the third form of the enzyme, HO-3 (McCoubrey et al., 1997b), are likely to insure the widespread interest in the enzyme system in the coming years. The present review is intended to highlight molecular properties of HO isozymes and their likely functions in the brain. Extended reviews of the system are found in Maines (1992, 1997).

  9. Developmental expression of heme oxygenase in the rat lung. (United States)

    Dennery, Phyllis A; Lee, Christen S; Ford, Berendera S; Weng, Yi-Hao; Yang, Guang; Rodgers, Pamela A


    Heme oxygenase (HO), the rate-limiting enzyme in the formation of bilirubin, is expressed in the lung and may serve as an antioxidant. This enzyme results in the formation of antioxidant bile pigments and the degradation of pro-oxidant heme. We wanted to evaluate the differences in expression of HO-1, the inducible form, and HO-2, the constitutive isoenzyme, during lung maturation and document whether lung HO expression was similar to that of other antioxidant enzymes. Lung total HO activity and HO-1 and HO-2 proteins as well as HO-1 and HO-2 mRNA were evaluated in animals from 16 d of gestation (e(16.5)) to 2 mo of age. Heme content was also evaluated because heme is the substrate of the reaction. HO-1 mRNA was maximal at e(19.5) and e(20.5), whereas HO-2 mRNA was not changed throughout maturation. Lung HO-1 protein was highest on the first days of life and lowest in adults, whereas HO-2 protein was maximally expressed at postnatal d 5 and then declined to reach adult values. As to HO activity, there was a prenatal peak at e(20.5), a second lesser peak at d 5, and thereafter a decline to adult values. Lung heme content was inversely correlated with HO activity or protein as the highest heme values were seen in adults with the lowest HO activity. In response to hyperoxia, HO-1 mRNA was induced only in the adult lungs. A better understanding of the maturational regulation of lung HO will define a role for HO in newborns at risk for oxygen toxicity.

  10. Genetic analyses of heme oxygenase 1 (HMOX1) in different forms of pancreatitis

    NARCIS (Netherlands)

    Weis, S.; Jesinghaus, M.; Kovacs, P.; Schleinitz, D.; Schober, R.; Ruffert, C.; Herms, M.; Wittenburg, H.; Stumvoll, M.; Blüher, M.; Grützmann, R.; Schulz, H.U.; Keim, V.; Mössner, J.; Bugert, P.; Witt, H.; Drenth, J.P.H.; Krohn, K.; Rosendahl, J.


    BACKGROUND: Heme oxygenase 1 (HMOX1) is the rate limiting enzyme in heme degradation and a key regulator of inflammatory processes. In animal models the course of pancreatitis was ameliorated by up-regulation of HMOX1 expression. Additionally, carbon monoxide released during heme breakdown inhibited

  11. Heme uptake by Leishmania amazonensis is mediated by the transmembrane protein LHR1.

    Directory of Open Access Journals (Sweden)

    Chau Huynh

    Full Text Available Trypanosomatid protozoan parasites lack a functional heme biosynthetic pathway, so must acquire heme from the environment to survive. However, the molecular pathway responsible for heme acquisition by these organisms is unknown. Here we show that L. amazonensis LHR1, a homolog of the C. elegans plasma membrane heme transporter HRG-4, functions in heme transport. Tagged LHR1 localized to the plasma membrane and to endocytic compartments, in both L. amazonensis and mammalian cells. Heme deprivation in L. amazonensis increased LHR1 transcript levels, promoted uptake of the fluorescent heme analog ZnMP, and increased the total intracellular heme content of promastigotes. Conversely, deletion of one LHR1 allele reduced ZnMP uptake and the intracellular heme pool by approximately 50%, indicating that LHR1 is a major heme importer in L. amazonensis. Viable parasites with correct replacement of both LHR1 alleles could not be obtained despite extensive attempts, suggesting that this gene is essential for the survival of promastigotes. Notably, LHR1 expression allowed Saccharomyces cerevisiae to import heme from the environment, and rescued growth of a strain deficient in heme biosynthesis. Syntenic genes with high sequence identity to LHR1 are present in the genomes of several species of Leishmania and also Trypanosoma cruzi and Trypanosoma brucei, indicating that therapeutic agents targeting this transporter could be effective against a broad group of trypanosomatid parasites that cause serious human disease.

  12. XAFS Debye-Waller factors for deformed hemes and metal substituted hemes

    Energy Technology Data Exchange (ETDEWEB)

    Dimakis, N; Mion, T; Ramirez, C [Department of Physics and Geology, University of Texas-Pan American, Edinburg, TX 78539 (United States); Bunker, G, E-mail: dimakis@utpa.ed [Department of Biological Chemical and Physical Sciences, Illinois Institute of Technology, Chicago, IL 60616 (United States)


    We present an efficient and accurate method for calculating XAFS Debye-Waller factors for deformed active sites of hemoproteins and metal substituted hemes. Based on the Normal Coordinate Structural Decomposition scheme, the deformation of the porphyrin macrocycle is expressed as a linear combination of the normal modes of the planar species. In our approach, we identify the modes that contribute most to the deformation. Small metal-porphyrin structures which match the macrocycle structural deformation of the deformed hemoprotein site are used to calculate the Debye-Waller parameters at sample's temperature. The Debye-Waller factors are directly obtained by calculating the normal mode spectrum of the corresponding metal-porphyrin structure using Density Functional Theory. Our method is tested on Ni-tetraadamantyl porphyrin and cytochrome c structures with more than 500 available scattering paths.

  13. Heme oxygenase-1 system and gastrointestinal inflammation: a short review. (United States)

    Zhu, Xiao; Fan, Wen-Guo; Li, Dong-Pei; Kung, Hsiangfu; Lin, Marie Cm


    Heme oxygenase-1 (HO-1) system catalyzes heme to biologically active products: carbon monoxide, biliverdin/bilirubin and free iron. It is involved in maintaining cellular homeostasis and many physiological and pathophysiological processes. A growing body of evidence indicates that HO-1 activation may play an important protective role in acute and chronic inflammation of gastrointestinal tract. This review focuses on the current understanding of the physiological significance of HO-1 induction and its possible roles in the gastrointestinal inflammation studied to date. The ability to upregulate HO-1 by pharmacological means or using gene therapy may offer therapeutic strategies for gastrointestinal inflammation in the future.

  14. Nanoparticles and self-organisation: the emergence of hierarchical properties from the nanoparticle soup (i.e., the small is getting bigger). Concluding remarks for Faraday Discussion: Nanoparticle Synthesis and Assembly. (United States)

    Schiffrin, David J


    Some four years ago, one of the participants in this Discussion (Prof. Nicholas Kotov) predicted that: "within five years we shall see multiple examples of electronic, sensor, optical and other devices utilizing self-assembled superstructures" (N. A. Kotov, J. Mater. Chem., 2011, 21, 16673-16674). Although this prediction came partially to fruition, we have witnessed an unprecedented interest in the properties of materials at the nanoscale. The point highlighted by Kotov, however, was the importance of self-assembly of structures from well characterised building blocks to yield hierarchical structures, hopefully with predictable properties, a concept that is an everyday pursuit of synthetic chemists. This Discussion has brought together researchers from a wide range of disciplines, i.e., colloid science, modelling, nanoparticle synthesis and organisation, magnetic and optical materials, and new imaging methods, within the excellent traditional Faraday Discussion format, to discuss advances in areas relevant to the main theme of the meeting.

  15. Analysis of Heme Iron Coordination in DGCR8: The Heme-Binding Component of the Microprocessor Complex. (United States)

    Girvan, Hazel M; Bradley, Justin M; Cheesman, Myles R; Kincaid, James R; Liu, Yilin; Czarnecki, Kazimierz; Fisher, Karl; Leys, David; Rigby, Stephen E J; Munro, Andrew W


    DGCR8 is the RNA-binding partner of the nuclease Drosha. Their complex (the "Microprocessor") is essential for processing of long, primary microRNAs (pri-miRNAs) in the nucleus. Binding of heme to DGCR8 is essential for pri-miRNA processing. On the basis of the split Soret ultraviolet-visible (UV-vis) spectrum of ferric DGCR8, bis-thiolate sulfur (cysteinate, Cys(-)) heme iron coordination of DGCR8 heme iron was proposed. We have characterized DGCR8 heme ligation using the Δ276 DGCR8 variant and combined electron paramagnetic resonance (EPR), magnetic circular dichroism (MCD), electron nuclear double resonance, resonance Raman, and electronic absorption spectroscopy. These studies indicate DGCR8 bis-Cys heme iron ligation, with conversion from bis-thiolate (Cys(-)/Cys(-)) axial coordination in ferric DGCR8 to bis-thiol (CysH/CysH) coordination in ferrous DGCR8. Pri-miRNA binding does not perturb ferric DGCR8's optical spectrum, consistent with the axial ligand environment being separated from the substrate-binding site. UV-vis absorption spectra of the Fe(II) and Fe(II)-CO forms indicate discrete species exhibiting peaks with absorption coefficients substantially larger than those for ferric DGCR8 and that previously reported for a ferrous form of DGCR8. Electron-nuclear double resonance spectroscopy data exclude histidine or water as axial ligands for ferric DGCR8 and favor bis-thiolate coordination in this form. UV-vis MCD and near-infrared MCD provide data consistent with this conclusion. UV-vis MCD data for ferrous DGCR8 reveal features consistent with bis-thiol heme iron coordination, and resonance Raman data for the ferrous-CO form are consistent with a thiol ligand trans to the CO. These studies support retention of DGCR8 cysteine coordination upon reduction, a conclusion distinct from those of previous studies of a different ferrous DGCR8 isoform.

  16. Supramolecular assembly based on a heteropolyanion: Synthesis and crystal structure of Na3(H2O)6[Al(OH)6Mo6O18]$\\cdot$2H2O

    Indian Academy of Sciences (India)

    Vaddypally Shivaiah; Samar K Das


    Synthesis and structural characterization of a polyoxometalate compound Na3(H2O)6 [Al(OH)6Mo6O18]$\\cdot$2H2O (1) have been described. Compound 1 exhibits three-dimensional network structure in the solid state, which is assembled by Anderson-type heteropolyanions, [Al(OH)6Mo6O18]$_n^{3n-}$, as building blocks sharing sodium cations. 1 possesses ``sinuous" channels occupied by supramolecular water dimers as guests. Anderson anions, sodium-coordinated water and crystal water are additionally involved in an intricate hydrogen-bonding network in the crystal of 1.

  17. Malaria parasite-synthesized heme is essential in the mosquito and liver stages and complements host heme in the blood stages of infection.

    Directory of Open Access Journals (Sweden)

    Viswanathan Arun Nagaraj

    Full Text Available Heme metabolism is central to malaria parasite biology. The parasite acquires heme from host hemoglobin in the intraerythrocytic stages and stores it as hemozoin to prevent free heme toxicity. The parasite can also synthesize heme de novo, and all the enzymes in the pathway are characterized. To study the role of the dual heme sources in malaria parasite growth and development, we knocked out the first enzyme, δ-aminolevulinate synthase (ALAS, and the last enzyme, ferrochelatase (FC, in the heme-biosynthetic pathway of Plasmodium berghei (Pb. The wild-type and knockout (KO parasites had similar intraerythrocytic growth patterns in mice. We carried out in vitro radiolabeling of heme in Pb-infected mouse reticulocytes and Plasmodium falciparum-infected human RBCs using [4-(14C] aminolevulinic acid (ALA. We found that the parasites incorporated both host hemoglobin-heme and parasite-synthesized heme into hemozoin and mitochondrial cytochromes. The similar fates of the two heme sources suggest that they may serve as backup mechanisms to provide heme in the intraerythrocytic stages. Nevertheless, the de novo pathway is absolutely essential for parasite development in the mosquito and liver stages. PbKO parasites formed drastically reduced oocysts and did not form sporozoites in the salivary glands. Oocyst production in PbALASKO parasites recovered when mosquitoes received an ALA supplement. PbALASKO sporozoites could infect mice only when the mice received an ALA supplement. Our results indicate the potential for new therapeutic interventions targeting the heme-biosynthetic pathway in the parasite during the mosquito and liver stages.

  18. Sequence assembly

    DEFF Research Database (Denmark)

    Scheibye-Alsing, Karsten; Hoffmann, S.; Frankel, Annett Maria


    Despite the rapidly increasing number of sequenced and re-sequenced genomes, many issues regarding the computational assembly of large-scale sequencing data have remain unresolved. Computational assembly is crucial in large genome projects as well for the evolving high-throughput technologies and...... in genomic DNA, highly expressed genes and alternative transcripts in EST sequences. We summarize existing comparisons of different assemblers and provide a detailed descriptions and directions for download of assembly programs at:

  19. Heme oxygenase-1 and carbon monoxide in pulmonary medicine. (United States)

    Slebos, Dirk-Jan; Ryter, Stefan W; Choi, Augustine M K


    Heme oxygenase-1 (HO-1), an inducible stress protein, confers cytoprotection against oxidative stress in vitro and in vivo. In addition to its physiological role in heme degradation, HO-1 may influence a number of cellular processes, including growth, inflammation, and apoptosis. By virtue of anti-inflammatory effects, HO-1 limits tissue damage in response to proinflammatory stimuli and prevents allograft rejection after transplantation. The transcriptional upregulation of HO-1 responds to many agents, such as hypoxia, bacterial lipopolysaccharide, and reactive oxygen/nitrogen species. HO-1 and its constitutively expressed isozyme, heme oxygenase-2, catalyze the rate-limiting step in the conversion of heme to its metabolites, bilirubin IXalpha, ferrous iron, and carbon monoxide (CO). The mechanisms by which HO-1 provides protection most likely involve its enzymatic reaction products. Remarkably, administration of CO at low concentrations can substitute for HO-1 with respect to anti-inflammatory and anti-apoptotic effects, suggesting a role for CO as a key mediator of HO-1 function. Chronic, low-level, exogenous exposure to CO from cigarette smoking contributes to the importance of CO in pulmonary medicine. The implications of the HO-1/CO system in pulmonary diseases will be discussed in this review, with an emphasis on inflammatory states.

  20. Heme oxygenase-1 and carbon monoxide in pulmonary medicine

    Directory of Open Access Journals (Sweden)

    Choi Augustine MK


    Full Text Available Abstract Heme oxygenase-1 (HO-1, an inducible stress protein, confers cytoprotection against oxidative stress in vitro and in vivo. In addition to its physiological role in heme degradation, HO-1 may influence a number of cellular processes, including growth, inflammation, and apoptosis. By virtue of anti-inflammatory effects, HO-1 limits tissue damage in response to proinflammatory stimuli and prevents allograft rejection after transplantation. The transcriptional upregulation of HO-1 responds to many agents, such as hypoxia, bacterial lipopolysaccharide, and reactive oxygen/nitrogen species. HO-1 and its constitutively expressed isozyme, heme oxygenase-2, catalyze the rate-limiting step in the conversion of heme to its metabolites, bilirubin IXα, ferrous iron, and carbon monoxide (CO. The mechanisms by which HO-1 provides protection most likely involve its enzymatic reaction products. Remarkably, administration of CO at low concentrations can substitute for HO-1 with respect to anti-inflammatory and anti-apoptotic effects, suggesting a role for CO as a key mediator of HO-1 function. Chronic, low-level, exogenous exposure to CO from cigarette smoking contributes to the importance of CO in pulmonary medicine. The implications of the HO-1/CO system in pulmonary diseases will be discussed in this review, with an emphasis on inflammatory states.

  1. Heme and menaquinone induced electron transport in lactic acid bacteria

    NARCIS (Netherlands)

    Brooijmans, R.J.W.; Smit, B.; Santos, dos F.; Riel, van J.; Vos, de W.M.; Hugenholtz, J.


    ABSTRACT: BACKGROUND: For some lactic acid bacteria higher biomass production as a result of aerobic respiration has been reported upon supplementation with heme and menaquinone. In this report, we have studied a large number of species among lactic acid bacteria for the existence of this trait. RES

  2. Heme and HO-1 inhibition of HCV, HBV, and HIV

    Directory of Open Access Journals (Sweden)

    Warren N Schmidt


    Full Text Available Hepatitis C virus, human immunodeficiency virus, and hepatitis B virus are chronic viral infections that cause considerable morbidity and mortality throughout the world. In the decades following the identification and sequencing of these viruses, in vitro experiments demonstrated that heme oxygenase-1, its oxidative products, and related compounds of the heme oxygenase system are virucidal for all three viruses. The purpose of this review is to critically evaluate and summarize the seminal studies that described and characterized this remarkable behavior. It will also discuss more recent work that discovered the antiviral mechanisms and target sites of these unique antiviral agents. In spite of the fact that these viruses are diverse pathogens with quite profound differences in structure and life cycle, it is significant that heme and related compounds show striking similarity for viral target sites across all three species. Collectively, these findings strongly indicate that we should move forward and develop heme and related tetrapyrroles into versatile antiviral agents that could be used therapeutically in patients with single or multiple viral infections.

  3. The Heme Connection: Linking Erythrocytes and Macrophage Biology (United States)

    Alam, Md Zahidul; Devalaraja, Samir; Haldar, Malay


    Erythroid function and development is intimately linked to macrophages. The primary function of erythrocytes is oxygen delivery, which is mediated by iron-containing hemoglobin. The major source of this iron is a recycling pathway where macrophages scavenge old and damaged erythrocytes to release iron contained within the heme moiety. Macrophages also promote erythropoiesis by providing a supportive niche in the bone marrow as an integral component of “erythorblastic islands.” Importantly, inflammation leads to alterations in iron handling by macrophages with significant impact on iron homeostasis and erythropoiesis. The importance of macrophages in erythropoiesis and iron homeostasis is well established and has been extensively reviewed. However, this developmental relationship is not one way, and erythrocytes can also regulate macrophage development and function. Erythrocyte-derived heme can induce the development of iron-recycling macrophages from monocytes, engage pattern recognition receptors to activate macrophages, and act as ligand for specific nuclear receptors to modulate macrophage function. Here, we discuss the role of heme as a signaling molecule impacting macrophage homeostasis. We will review these actions of heme within the framework of our current understanding of the role of micro-environmental factors in macrophage development and function. PMID:28167947

  4. Mechanism of reaction of chlorite with mammalian heme peroxidases. (United States)

    Jakopitsch, Christa; Pirker, Katharina F; Flemmig, Jörg; Hofbauer, Stefan; Schlorke, Denise; Furtmüller, Paul G; Arnhold, Jürgen; Obinger, Christian


    This study demonstrates that heme peroxidases from different superfamilies react differently with chlorite. In contrast to plant peroxidases, like horseradish peroxidase (HRP), the mammalian counterparts myeloperoxidase (MPO) and lactoperoxidase (LPO) are rapidly and irreversibly inactivated by chlorite in the micromolar concentration range. Chlorite acts as efficient one-electron donor for Compound I and Compound II of MPO and LPO and reacts with the corresponding ferric resting states in a biphasic manner. The first (rapid) phase is shown to correspond to the formation of a MPO-chlorite high-spin complex, whereas during the second (slower) phase degradation of the prosthetic group was observed. Cyanide, chloride and hydrogen peroxide can block or delay heme bleaching. In contrast to HRP, the MPO/chlorite system does not mediate chlorination of target molecules. Irreversible inactivation is shown to include heme degradation, iron release and decrease in thermal stability. Differences between mammalian peroxidases and HRP are discussed with respect to differences in active site architecture and heme modification.

  5. Structural and spectroscopic characterisation of a heme peroxidase from sorghum. (United States)

    Nnamchi, Chukwudi I; Parkin, Gary; Efimov, Igor; Basran, Jaswir; Kwon, Hanna; Svistunenko, Dimitri A; Agirre, Jon; Okolo, Bartholomew N; Moneke, Anene; Nwanguma, Bennett C; Moody, Peter C E; Raven, Emma L


    A cationic class III peroxidase from Sorghum bicolor was purified to homogeneity. The enzyme contains a high-spin heme, as evidenced by UV-visible spectroscopy and EPR. Steady state oxidation of guaiacol was demonstrated and the enzyme was shown to have higher activity in the presence of calcium ions. A Fe(III)/Fe(II) reduction potential of -266 mV vs NHE was determined. Stopped-flow experiments with H2O2 showed formation of a typical peroxidase Compound I species, which converts to Compound II in the presence of calcium. A crystal structure of the enzyme is reported, the first for a sorghum peroxidase. The structure reveals an active site that is analogous to those for other class I heme peroxidase, and a substrate binding site (assigned as arising from binding of indole-3-acetic acid) at the γ-heme edge. Metal binding sites are observed in the structure on the distal (assigned as a Na(+) ion) and proximal (assigned as a Ca(2+)) sides of the heme, which is consistent with the Ca(2+)-dependence of the steady state and pre-steady state kinetics. It is probably the case that the structural integrity (and, thus, the catalytic activity) of the sorghum enzyme is dependent on metal ion incorporation at these positions.

  6. Heme oxygenase-1/carbon monoxide: from metabolism to molecular therapy. (United States)

    Ryter, Stefan W; Choi, Augustine M K


    Heme oxygenase-1 (HO-1), a ubiquitous inducible stress-response protein, serves a major metabolic function in heme turnover. HO activity cleaves heme to form biliverdin-IXalpha, carbon monoxide (CO), and iron. Genetic experiments have revealed a central role for HO-1 in tissue homeostasis, protection against oxidative stress, and in the pathogenesis of disease. Four decades of research have witnessed not only progress in elucidating the molecular mechanisms underlying the regulation and function of this illustrious enzyme, but also have opened remarkable translational applications for HO-1 and its reaction products. CO, once regarded as a metabolic waste, can act as an endogenous mediator of cellular signaling and vascular function. Exogenous application of CO by inhalation or pharmacologic delivery can confer cytoprotection in preclinical models of lung/vascular injury and disease, based on anti-apoptotic, anti-inflammatory, and anti-proliferative properties. The bile pigments, biliverdin and bilirubin, end products of heme degradation, have also shown potential as therapeutics in vascular disease based on anti-inflammatory and anti-proliferative activities. Further translational and clinical trials research will unveil whether the HO-1 system or any of its reaction products can be successfully applied as molecular medicine in human disease.

  7. Degradation of Heme in Gram-Negative Bacteria: the Product of the hemO Gene of Neisseriae Is a Heme Oxygenase



    A full-length heme oxygenase gene from the gram-negative pathogen Neisseria meningitidis was cloned and expressed in Escherichia coli. Expression of the enzyme yielded soluble catalytically active protein and caused accumulation of biliverdin within the E. coli cells. The purified HemO forms a 1:1 complex with heme and has a heme protein spectrum similar to that previously reported for the purified heme oxygenase (HmuO) from the gram-positive pathogen Corynebacterium diphtheriae and for eukar...

  8. Mechanism of horseradish peroxidase-catalyzed heme oxidation and polymerization (beta-hematin formation). (United States)

    Trivedi, Vishal; Chand, Prem; Maulik, Prakas R; Bandyopadhyay, Uday


    Horseradish peroxidase (HRP) catalyzes the polymerization of free heme (beta-hematin formation) through its oxidation. Heme when added to HRP compound II (FeIV=O) causes spectral shift from 417 nm (Compound II) to 402 nm (native, FeIII) indicating that heme may be oxidized via one-electron transfer. Direct evidence for one-electron oxidation of heme by HRP intermediates is provided by the appearance of an E.s.r signal of a 5,5-dimethyl-1-pyrroline N-oxide (spin trap)-heme radical adduct (a1H=14.75 G, a2H=4.0 G) in E.s.r studies. Heme-polymerization by HRP is inhibited by spin trap indicating that one-electron oxidation product of heme ultimately leads to the formation of heme-polymer. HRP, when incubated with diethyl pyrocarbonate (DEPC), a histidine specific reagent, shows concentration dependent loss of heme-polymerization indicating the role of histidine residues in the process. We suggest that HRP catalyzes the formation of heme-polymer through one-electron oxidation of free heme.

  9. Haemoproteus and Schistosoma synthesize heme polymers similar to Plasmodium hemozoin and beta-hematin. (United States)

    Chen, M M; Shi, L; Sullivan, D J


    Many parasites digest hemoglobin as an amino acid source, but only a few produce heme polymer pigment instead of catabolizing heme via heme oxygenase. This work compares purified heme polymers produced by Haemoproteus columbae and Schistosoma mansoni to that of Plasmodium falciparum hemozoin and synthetic beta-hematin. Fourier-transform infrared spectroscopy identifies the signature peaks of the common iron-carboxylate bond characteristic in all four heme polymers. However, all pigments could be distinguished by quite different three-dimensional structure visualized by Field Emission Inlens Scanning Electron Microscopy. Both P. falciparum and H. columbae heme polymers had a symmetrical shape unlike the amorphous S. mansoni heme polymer and beta-hematin. All four heme pigments serve as templates for heme polymer extension, which was inhibitable by chloroquine and other quinoline antimalarials. The polymers showed different levels of resistance to hydrogen peroxide degradation. This work identifies another genus, Haemoproteus, capable of intracellular heme polymer formation. The different three-dimensional structures of each pigment implicate genus specific formation of heme polymer, variation of inhibition of polymer extension by the quinolines and degradation by hydrogen peroxide.

  10. Identification and characterization of a heme periplasmic-binding protein in Haemophilus ducreyi. (United States)

    St Denis, Melissa; Sonier, Brigitte; Robinson, Renée; Scott, Fraser W; Cameron, D William; Lee, B Craig


    Haemophilus ducreyi, a gram-negative and heme-dependent bacterium, is the causative agent of chancroid, a genital ulcer sexually transmitted infection. Heme acquisition in H. ducreyi proceeds via a receptor mediated process in which the initial event involves binding of hemoglobin and heme to their cognate outer membrane proteins, HgbA and TdhA, respectively. Following this specific interaction, the fate of the periplasmic deposited heme is unclear. Using protein expression profiling of the H. ducreyi periplasmic proteome, a periplasmic-binding protein, termed hHbp, was identified whose expression was enhanced under heme-limited conditions. The gene encoding this protein was situated in a locus displaying genetic characteristics of an ABC transporter. The purified protein bound heme in a dose-dependent and saturable manner and this binding was specifically competitively inhibited by heme. The hhbp gene functionally complemented an Escherichia coli heme uptake mutant. Expression of the heme periplasmic-binding protein was detected in a limited survey of H. ducreyi and H. influenzae clinical strains. These results indicate that the passage of heme into the cytoplasm of H. ducreyi involves a heme dedicated ABC transporter.

  11. Structural basis of heme binding in the Cu,Zn superoxide dismutase from Haemophilus ducreyi. (United States)

    Töro, Imre; Petrutz, Cristiana; Pacello, Francesca; D'Orazio, Melania; Battistoni, Andrea; Djinović-Carugo, Kristina


    The Cu,Zn superoxide dismutase from Haemophilus ducreyi is characterized by the unique ability to bind heme at its dimer interface. Here we report the high-resolution crystal structures of this protein in the heme-loaded (holo) and heme-free (apo) forms. Heme is asymmetrically bound between the two enzyme subunits, where heme iron is coordinated by two histidine residues, His64 and His 124, provided by the two subunits. Moreover, the binding of heme to the protein is ensured by stabilizing contacts between the prosthetic group and a limited number of other residues, most of which are not present in other bacterial enzyme variants. We show that the introduction of only three mutations at the dimer interface of the enzyme from Haemophilus parainfluenzae, a closely related bacterial species, is sufficient to induce heme-binding ability by this enzyme variant. Heme binding does not alter protein activity. Moreover, the binding of the prosthetic group does not induce any significant structural perturbation at the subunit level and requires only limited local structural rearrangements that widen the cleft at the dimer interface and cause a limited shift in the relative orientation between the subunits. The presence of a preformed heme-binding pocket and the significant solvent exposure of the cofactor to the solvent are compatible with the suggested protective role of the enzyme against heme toxicity or with its involvement in heme trafficking in the periplasmic space.

  12. PREFACE: IUMRS-ICA 2008 Symposium, Sessions 'X. Applications of Synchrotron Radiation and Neutron Beam to Soft Matter Science' and 'Y. Frontier of Polymeric Nano-Soft-Materials - Precision Polymer Synthesis, Self-assembling and Their Functionalization' (United States)

    Takahara, Atsushi; Kawahara, Seiichi


    Tashiro (Toyota Technological Institute) Professor Kazuo Sakurai(Kitakyushu University) Professor Keiji Tanaka (Kyushu University) Dr Sono Sasaki (JASRI/Spring-8) Professor Naoya Torikai (KENS) Professor Moonhor Ree (POSTECH) Professor Kookheon Char (Seoul National University) Professor Charles C Han (CAS) Professor Atsushi Takahara(Kyushu University) Frontier of Polymeric Nano-Soft-Materials, Precision Polymer Synthesis, Self-assembling and Their Functionalization (Symposium Y of IUMRS-ICA2008) Seiichi Kawahara, Rong-Ming Ho, Hiroshi Jinnai, Masami Kamigaito, Takashi Miyata, Hiroshi Morita, Hideyuki Otsuka, Daewon Sohn, Keiji Tanaka It is our great pleasure and honor to publish peer-reviewed papers, presented in Symposium Y 'Frontier of Polymeric Nano-Soft-Materials Precision Polymer Synthesis, Self-assembling and Their Functionalization' at the International Union of Materials Research Societies International Conference in Asia 2008 (IUMRS-ICA2008), which was held on 9-13 December 2008, at Nagoya Congress Center, Nagoya, Japan. 'Polymeric nano-soft-materials' are novel outcomes based on a recent innovative evolution in polymer science, i.e. precision polymer synthesis, self-assembling and functionalization of multi-component systems. The materials are expected to exhibit specific functions and unique properties due to their hierarchic morphologies brought either by naturally-generated ordering or by artificial manipulation of the systems, e.g., crystallization and phase-separation. The emerging precision synthesis has brought out new types of polymers with well-controlled primary structures. Furthermore, the surface and interface of the material are recognized to play an important role in the outstanding mechanical, electrical and optical properties, which are required for medical and engineering applications. In order to understand structure-property relationships in the nano-soft-materials, it is indispensable to develop novel characterization techniques. Symposium Y

  13. Regulatory Fe(II/III) heme: the reconstruction of a molecule's biography. (United States)

    Kühl, Toni; Imhof, Diana


    More than 20 years of research on heme as a temporary effector molecule of proteins have revealed its widespread impact on virtually all primary functions in the human organism. As our understanding of this influence is still growing, a comprehensive overview of compiled data will give fresh impetus for creativity and developing new strategies in heme-related research. From known data concerning heme-regulated proteins and their involvement in the development of diseases, we provide concise information of Fe(II/III) heme as a regulator and the availability of "regulatory heme". The latter is dependent on the balance between free and bound Fe(II/III) heme, here termed "hemeostasis". Imbalance of this system can lead to the development of diseases that were not always attributed to this small molecule. Diseases such as cancer or Alzheimer's disease highlight the reawakened interest in heme, whose function was previously believed to be completely understood.

  14. Dietary heme-mediated PPARa activation does not affect the heme-induced epithelial hyperproliferation and hyperplasia in mouse colon

    NARCIS (Netherlands)

    IJssenagger, N.; Wit, de N.J.W.; Muller, M.R.; Meer, van der R.


    Red meat consumption is associated with an increased colon cancer risk. Heme, present in red meat, injures the colon surface epithelium by luminal cytotoxicity and reactive oxygen species. This surface injury is overcompensated by hyperproliferation and hyperplasia of crypt cells. Transcriptome anal

  15. Differential induction of heme oxygenase and other stress proteins in cultured hippocampal astrocytes and neurons by inorganic lead. (United States)

    Cabell, Leigh; Ferguson, Charles; Luginbill, Deana; Kern, Marcey; Weingart, Adam; Audesirk, Gerald


    We examined the effects of exposure to inorganic lead (Pb2+) on the induction of stress proteins in cultured hippocampal neurons and astrocytes, with particular emphasis on the induction of heme oxygenase-1 (HO-1). In radiolabeled neuronal cultures, Pb2+ exposure had no significant effect on the synthesis of any protein at any concentration (up to 250 microM) or duration of exposure (up to 4 days). In radiolabeled astrocyte cultures, however, Pb2+ exposure (100 nM to 100 microM; 1-4 days) increased synthesis of proteins with approximate molecular weights of 23, 32, 45, 57, 72, and 90 kDa. Immunoblot experiments showed that Pb2+ exposure (100 nM to 10 microM, 1-14 days) induces HO-1 synthesis in astrocytes, but not in neurons; this is probably the 32-kDa protein. The other heme oxygenase isoform, HO-2, is present in both neurons and astrocytes, but is not inducible by Pb2+ at concentrations up to 100 microM. HO-1 can be induced by a variety of stimuli. We found that HO-1 induction in astrocytes is increased by combined exposure to Pb2+ and many other stresses, including heat, nitric oxide, H2O2, and superoxide. One of the stimuli that may induce HO-1 is oxidative stress. Lead exposure causes oxidative stress in many cell types, including astrocytes. Induction of HO-1 by Pb2+ is reduced by the hydroxyl radical scavengers dimethylthiourea (DMTU) and mannitol, but not by inhibitors of calmodulin, calmodulin-dependent protein kinases, protein kinase C, or extracellular signal-regulated kinases (ERK). Therefore, we conclude that oxidative stress is an important mechanism by which Pb2+ induces HO-1 synthesis in astrocytes.

  16. Enzymes of heme metabolism in the kidney: regulation by trace metals which do not form heme complexes. (United States)

    Maines, M D; Kappas, A


    The in vivo regulation by metal ions of the enzymes of heme metabolism in kidney-particularly of ALAS, the rate-limiting enzyme in heine formation- was investigated. Ni(2+) and Pt(4+), metals which do not enzymatically form metalloporphyrins, were found to regulate ALAS in kidney as they do in liver. The pattern of this regulation was generally similar to that observed with heme and metal ions in liver, i.e., a late increase in enzyme activity after an early period in which ALAS activity was unaltered or inhibited. The metals did not interact with the enzyme in vitro to alter its activity. In this study no direct reciprocal relationship between ALAS activity and total cellular heine content was demonstrated. The metal ions, particularly Pt(4+), also altered the activity of other enzymes of heme biosynthesis in kidney. Pt(4+) severely inhibited the activity of ALAD and UROS. Ni(2+) and Pt(4+) were potent inducers of heme oxygenase, the initial and rate-limiting enzyme in heine degradation. It is proposed that the physiological regulation of ALAS is mediated through the action of metal ions, rather than by the cellular content of heine, and that the regulation of ALAS by heine reflects the action of the central metal ion of heme rather than that of the entire metalloporphyrin complex. In this proposed mechanism for metal ion regulation of ALAS, the tetrapyrrole moiety of heine is considered to function principally as an efficient carrier of metal to the regulatory site for ALAS production, inasmuch as the tetrapyrrole ring itself has been shown in earlier studies not to have any effect on ALAS activity. The production of heine oxygenase is believed to be similarly regulated.

  17. Macrophage preconditioning with synthetic malaria pigment reduces cytokine production via heme iron-dependent oxidative stress. (United States)

    Taramelli, D; Recalcati, S; Basilico, N; Olliaro, P; Cairo, G


    Hemozoin (malaria pigment), a polymer of hematin (ferri-protoporphyrin IX) derived from hemoglobin ingested by intraerythrocytic plasmodia, modulates cytokine production by phagocytes. Mouse peritoneal macrophages (PM) fed with synthetic beta-hematin (BH), structurally identical to native hemozoin, no longer produce tumor necrosis factor alpha (TNFalpha) and nitric oxide (NO) in response to lipopolysaccharide (LPS). Impairment of NO synthesis is due to inhibition of inducible nitric oxide synthase (iNOS) production. BH-mediated inhibition of PM functions cannot be ascribed to iron release from BH because neither prevention by iron chelators nor down-regulation of iron-regulatory protein activity was detected. Inhibition appears to be related to pigment-induced oxidative stress because (a) thiol compounds partially restored PM functions, (b) heme oxygenase (HO-1) and catalase mRNA levels were up-regulated, and (c) free radicals production increased in BH-treated cells. The antioxidant defenses of the cells determine the response to BH: microglia cells, which show a lower extent of induction of HO-1 and catalase mRNAs and lower accumulation of oxygen radicals, are less sensitive to the inhibitory effect of BH on cytokine production. Results indicate that BH is resistant to degradation by HO-1 and that heme-iron mediated oxidative stress may contribute to malaria-induced immunosuppression. This study may help correlate the different clinical manifestations of malaria, ranging from uncomplicated to severe disease, with dysregulation of phagocyte functions and promote better therapeutic strategies to counteract the effects of hemozoin accumulation.

  18. Heme oxygenase-1: redox regulation and role in the hepatic response to oxidative stress. (United States)

    Bauer, Michael; Bauer, Inge


    Heme oxygenase (HO) catalyzes the oxidative cleavage of the alpha-mesocarbon of Fe-protoporphyrin-IX yielding equimolar amounts of biliverdin-IXalpha, free divalent iron, and carbon monoxide (CO). Among the three isoenzymes cloned to date, only HO-1 can be induced by a variety of seemingly disparate stimuli, most of which are linked by their ability to provoke oxidative stress. Although constitutive expression of HO-1 in the liver is restricted to Kupffer cells, the gene is inducible in nonparenchymal as well as in parenchymal liver cells. HO-1 induction potentially confers protection against oxidative stress in a variety of experimental models, such as liver ischemia/reperfusion secondary to transplantation or hemorrhage/resuscitation. Induction of HO-1 may protect the cell against oxidative injury by (a) controlling intracellular levels of "free" heme (a prooxidant), (b) producing biliverdin (an antioxidant), (c) improving nutritive perfusion via CO release, and (d) fostering the synthesis of the Fe-binding protein ferritin. Although protective effects of up-regulation of the HO pathway--presumably through production of bile pigments and CO--have been reported for a variety of cells and tissues, including the liver, evidence suggests that the protective action might be restricted to a rather narrow threshold of overexpression. High levels of HO-1 may even sensitize the cell to oxidative stress, e.g., through release of reactive iron. Transcriptional activation of the HO-1 gene is an integral part of the cellular response to oxidative stress, but its induction seems to be neither exclusively cytoprotective nor exclusively cytotoxic.

  19. Combination of ionic self-assembly and hydrogen bonding as a tool for the synthesis of liquid-crystalline materials and organogelators from a simple building block. (United States)

    Camerel, Franck; Faul, Charl F J


    In this communication we report on the facile combination of hydrogen bonding and the ionic self-assembly (ISA) process to produce organized materials and fiber-containing organogel superstructures from functionalised oligoelectrolytic building blocks.

  20. The dppBCDF gene cluster of Haemophilus influenzae: Role in heme utilization

    Directory of Open Access Journals (Sweden)

    Morton Daniel J


    Full Text Available Abstract Background Haemophilus influenzae requires a porphyrin source for aerobic growth and possesses multiple mechanisms to obtain this essential nutrient. This porphyrin requirement may be satisfied by either heme alone, or protoporphyrin IX in the presence of an iron source. One protein involved in heme acquisition by H. influenzae is the periplasmic heme binding protein HbpA. HbpA exhibits significant homology to the dipeptide and heme binding protein DppA of Escherichia coli. DppA is a component of the DppABCDF peptide-heme permease of E. coli. H. influenzae homologs of dppBCDF are located in the genome at a point distant from hbpA. The object of this study was to investigate the potential role of the H. influenzae dppBCDF locus in heme utilization. Findings An insertional mutation in dppC was constructed and the impact of the mutation on the utilization of both free heme and various proteinaceous heme sources as well as utilization of protoporphyrin IX was determined in growth curve studies. The dppC insertion mutant strain was significantly impacted in utilization of all tested heme sources and protoporphyin IX. Complementation of the dppC mutation with an intact dppCBDF gene cluster in trans corrected the growth defects seen in the dppC mutant strain. Conclusion The dppCBDF gene cluster constitutes part of the periplasmic heme-acquisition systems of H. influenzae.

  1. The Effect of Plant Proteins Derived from Cereals and Legumes on Heme Iron Absorption

    Directory of Open Access Journals (Sweden)

    Valerie Weinborn


    Full Text Available The aim of this study is to determine the effect of proteins from cereals and legumes on heme iron (Fe absorption. The absorption of heme Fe without its native globin was measured. Thirty adult females participated in two experimental studies (15 per study. Study I focused on the effects of cereal proteins (zein, gliadin and glutelin and study II on the effects of legume proteins (soy, pea and lentil on heme Fe absorption. When heme was given alone (as a control, study I and II yielded 6.2% and 11.0% heme absorption (p > 0.05. In study I, heme Fe absorption was 7.2%, 7.5% and 5.9% when zein, gliadin and glutelin were added, respectively. From this, it was concluded that cereal proteins did not affect heme Fe absorption. In study II, heme Fe absorption was 7.3%, 8.1% and 9.1% with the addition of soy, pea and lentil proteins, respectively. Only soy proteins decreased heme Fe absorption (p < 0.05. These results suggest that with the exception of soy proteins, which decreased absorption, proteins derived from cereals and legumes do not affect heme Fe absorption.

  2. The Effect of Plant Proteins Derived from Cereals and Legumes on Heme Iron Absorption. (United States)

    Weinborn, Valerie; Pizarro, Fernando; Olivares, Manuel; Brito, Alex; Arredondo, Miguel; Flores, Sebastián; Valenzuela, Carolina


    The aim of this study is to determine the effect of proteins from cereals and legumes on heme iron (Fe) absorption. The absorption of heme Fe without its native globin was measured. Thirty adult females participated in two experimental studies (15 per study). Study I focused on the effects of cereal proteins (zein, gliadin and glutelin) and study II on the effects of legume proteins (soy, pea and lentil) on heme Fe absorption. When heme was given alone (as a control), study I and II yielded 6.2% and 11.0% heme absorption (p > 0.05). In study I, heme Fe absorption was 7.2%, 7.5% and 5.9% when zein, gliadin and glutelin were added, respectively. From this, it was concluded that cereal proteins did not affect heme Fe absorption. In study II, heme Fe absorption was 7.3%, 8.1% and 9.1% with the addition of soy, pea and lentil proteins, respectively. Only soy proteins decreased heme Fe absorption (p legumes do not affect heme Fe absorption.

  3. High throughput heme assay by detection of chemiluminescence of reconstituted horseradish peroxidase. (United States)

    Takahashi, Shigekazu; Masuda, Tatsuru


    In living organisms, heme is an essential molecule for various biological functions. Recent studies also suggest that heme functions as organelle-derived signal that regulates fundamental cell processes. Furthermore, estimation of heme is widely used for studying various blood disorders. In this regard, development of a rapid, sensitive, and high throughput heme assay has been sought. The most frequently used method of measuring heme by pyridine hemochrome is time, labor, and material intensive, and therefore limiting in its utility for large scale, high throughput analysis. Recently, we reported alternative method that is sensitive and specific to heme, which is based on the ability of horseradish peroxidase (HRP) apo-enzyme to reconstitute with heme to form an active holo-enzyme. Here, we developed high throughput heme assay by performing reactions on multi-well plate with highly sensitive chemiluminescence detection reagents. Detection of chemiluminescence in charged coupled device (CCD)-based gel doc apparatus enables simultaneous measurement of multiple samples. Furthermore, the high sensitivity of this assay allowed a direct measurement of heme in solvent extracts after dilution. This assay is sensitive, quick, provides a large dynamic range, and is well suited for large-scale analysis of heme extracted from minute amount of samples.

  4. Alteration by irradiation and storage at amount of heme iron in poultry meat; Alteracoes provocadas pela irradiacao e armazenamento nos teores de ferro heme em carne de frango

    Energy Technology Data Exchange (ETDEWEB)

    Souza, Adriana Regia Marques de; Arthur, Valter Arthur [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Lab. de Irradiacao de Alimentos e Radioentomologia; Canniatti-Brazaca, Solange Guidolin [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil). Dept. de Agroindustria, Alimentos e Nutricao]. E-mail:


    Studies of irradiation and storage effects in chicken were carried out to discover the influence in iron heme, non-heme amount, color and total pigments. Chicken thighs and chicken breast were studied. These were irradiated to 0, 1 and 2 kGy stored by 14 days to 4 deg C in refrigerator. Determining the heme content and non-heme of meat was done using the colorimeter method and the Ferrozine reagent. The values of iron heme were influenced both by the irradiation and the storage, reducing the amount throughout the course of time. The iron non-heme was also influenced by the doses and the storage time, however the values increased throughout the course of time, because of the conversion of iron heme in non-heme. The color did not show that it was influenced by the studied doses, except for the storage, and the total number of pigments was affected by the irradiation and the time, reducing the values with the increase of storage. Irradiation was shown to be a good method to conserve iron. (author)

  5. Oms1 associates with cytochrome c oxidase assembly intermediates to stabilize newly synthesized Cox1. (United States)

    Bareth, Bettina; Nikolov, Miroslav; Lorenzi, Isotta; Hildenbeutel, Markus; Mick, David U; Helbig, Christin; Urlaub, Henning; Ott, Martin; Rehling, Peter; Dennerlein, Sven


    The mitochondrial cytochrome c oxidase assembles in the inner membrane from subunits of dual genetic origin. The assembly process of the enzyme is initiated by membrane insertion of the mitochondria-encoded Cox1 subunit. During complex maturation, transient assembly intermediates, consisting of structural subunits and specialized chaperone-like assembly factors, are formed. In addition, cofactors such as heme and copper have to be inserted into the nascent complex. To regulate the assembly process, the availability of Cox1 is under control of a regulatory feedback cycle in which translation of COX1 mRNA is stalled when assembly intermediates of Cox1 accumulate through inactivation of the translational activator Mss51. Here we isolate a cytochrome c oxidase assembly intermediate in preparatory scale from coa1Δ mutant cells, using Mss51 as bait. We demonstrate that at this stage of assembly, the complex has not yet incorporated the heme a cofactors. Using quantitative mass spectrometry, we define the protein composition of the assembly intermediate and unexpectedly identify the putative methyltransferase Oms1 as a constituent. Our analyses show that Oms1 participates in cytochrome c oxidase assembly by stabilizing newly synthesized Cox1.

  6. Challenging Density Functional Theory Calculations with Hemes and Porphyrins

    Directory of Open Access Journals (Sweden)

    Sam P. de Visser


    Full Text Available In this paper we review recent advances in computational chemistry and specifically focus on the chemical description of heme proteins and synthetic porphyrins that act as both mimics of natural processes and technological uses. These are challenging biochemical systems involved in electron transfer as well as biocatalysis processes. In recent years computational tools have improved considerably and now can reproduce experimental spectroscopic and reactivity studies within a reasonable error margin (several kcal·mol−1. This paper gives recent examples from our groups, where we investigated heme and synthetic metal-porphyrin systems. The four case studies highlight how computational modelling can correctly reproduce experimental product distributions, predicted reactivity trends and guide interpretation of electronic structures of complex systems. The case studies focus on the calculations of a variety of spectroscopic features of porphyrins and show how computational modelling gives important insight that explains the experimental spectra and can lead to the design of porphyrins with tuned properties.

  7. Heme oxygenase-1 deficiency: the first autopsy case. (United States)

    Kawashima, Atsuhiro; Oda, Yoshio; Yachie, Akihiro; Koizumi, Shoichi; Nakanishi, Isao


    This article describes the first autopsy case of heme oxygenase (HO)-1 deficiency. A 6-year-old boy who presented with growth retardation; anemia; leukocytosis; thrombocytosis; coagulation abnormality; elevated levels of haptoglobin, ferritin, and heme in serum; a low serum bilirubin concentration; and hyperlipidemia was diagnosed as HO-1 deficient by gene analysis several months before death. Autopsy showed amyloid deposits in the liver and adrenal glands and mesangioproliferative glomerular changes in kidneys, in addition to an irregular distribution of foamy macrophages with iron pigments. Fatty streaks and fibrous plaques were noted in the aorta. Compared with HO-1--targeted mice, the present case seems to more severely involve endothelial cells and the reticuloendothelial system, resulting in intravascular hemolysis, disseminated intravascular coagulation, and amyloidosis with a short survival. This contrasts to the predominant iron metabolic disorders of HO-1--targeted mice with a long survival.

  8. Heme oxygenase-1 induction by dieldrin in dopaminergic cells. (United States)

    Kim, Do Kyung; Kim, Jae-Sung; Kim, Ji-Eun; Kim, Sung-Jun; Lee, Jung-Sup; Kim, Dae-Joong; Son, Jin H; Chun, Hong Sung


    We investigated the transcriptional events and signaling pathways involved in the induction of heme oxygenase-1 (HO-1) by dieldrin, an environmental risk factor of Parkinson's disease, in a dopaminergic neuronal cells (SN4741). Dieldrin exposure caused dose-dependent and time-dependent induction of heme oxygenase activity and HO-1 protein expression. Deletional and mutational analyses showed that the 5' distal enhancers, E1 and E2, mediate dieldrin-induced HO-1 gene transcription, and the AP-1 DNA binding sites in the E2 enhancer are critical for E2-mediated HO-1 gene activation. Furthermore, both the p38 and JNK mitogen-activated protein kinase pathways are utilized for HO-1 transcriptional activation by dieldrin. HO-1 inhibitor, ZnPP IX reduced the expression of HO-1 but enhanced the cytotoxicity induced by dieldrin.

  9. Cyanide binding to human plasma heme-hemopexin: A comparative study

    Energy Technology Data Exchange (ETDEWEB)

    Ascenzi, Paolo, E-mail: [Laboratorio Interdipartimentale di Microscopia Elettronica, Universita Roma Tre, Roma (Italy); Istituto Nazionale di Biostrutture e Biosistemi, Roma (Italy); Leboffe, Loris [Istituto Nazionale di Biostrutture e Biosistemi, Roma (Italy); Polticelli, Fabio [Dipartimento di Biologia, Universita Roma Tre, Roma (Italy)


    Highlights: Black-Right-Pointing-Pointer Cyanide binding to ferric HHPX-heme-Fe. Black-Right-Pointing-Pointer Cyanide binding to ferrous HHPX-heme-Fe. Black-Right-Pointing-Pointer Dithionite-mediated reduction of ferric HHPX-heme-Fe-cyanide. Black-Right-Pointing-Pointer Cyanide binding to HHPX-heme-Fe is limited by ligand deprotonation. Black-Right-Pointing-Pointer Cyanide dissociation from HHPX-heme-Fe-cyanide is limited by ligand protonation. -- Abstract: Hemopexin (HPX) displays a pivotal role in heme scavenging and delivery to the liver. In turn, heme-Fe-hemopexin (HPX-heme-Fe) displays heme-based spectroscopic and reactivity properties. Here, kinetics and thermodynamics of cyanide binding to ferric and ferrous hexa-coordinate human plasma HPX-heme-Fe (HHPX-heme-Fe(III) and HHPX-heme-Fe(II), respectively), and for the dithionite-mediated reduction of the HHPX-heme-Fe(III)-cyanide complex, at pH 7.4 and 20.0 Degree-Sign C, are reported. Values of thermodynamic and kinetic parameters for cyanide binding to HHPX-heme-Fe(III) and HHPX-heme-Fe(II) are K = (4.1 {+-} 0.4) Multiplication-Sign 10{sup -6} M, k{sub on} = (6.9 {+-} 0.5) Multiplication-Sign 10{sup 1} M{sup -1} s{sup -1}, and k{sub off} = 2.8 Multiplication-Sign 10{sup -4} s{sup -1}; and H = (6 {+-} 1) Multiplication-Sign 10{sup -1} M, h{sub on} = 1.2 Multiplication-Sign 10{sup -1} M{sup -1} s{sup -1}, and h{sub off} = (7.1 {+-} 0.8) Multiplication-Sign 10{sup -2} s{sup -1}, respectively. The value of the rate constant for the dithionite-mediated reduction of the HHPX-heme-Fe(III)-cyanide complex is l = 8.9 {+-} 0.8 M{sup -1/2} s{sup -1}. HHPX-heme-Fe reactivity is modulated by proton acceptor/donor amino acid residue(s) (e.g., His236) assisting the deprotonation and protonation of the incoming and outgoing ligand, respectively.

  10. Mosaic origin of the heme biosynthesis pathway in photosynthetic eukaryotes. (United States)

    Oborník, Miroslav; Green, Beverley R


    Heme biosynthesis represents one of the most essential metabolic pathways in living organisms, providing the precursors for cytochrome prosthetic groups, photosynthetic pigments, and vitamin B(12). Using genomic data, we have compared the heme pathway in the diatom Thalassiosira pseudonana and the red alga Cyanidioschyzon merolae to those of green algae and higher plants, as well as to those of heterotrophic eukaryotes (fungi, apicomplexans, and animals). Phylogenetic analyses showed the mosaic character of this pathway in photosynthetic eukaryotes. Although most of the algal and plant enzymes showed the expected plastid (cyanobacterial) origin, at least one of them (porphobilinogen deaminase) appears to have a mitochondrial (alpha-proteobacterial) origin. Another enzyme, glutamyl-tRNA synthase, obviously originated in the eukaryotic nucleus. Because all the plastid-targeted sequences consistently form a well-supported cluster, this suggests that genes were either transferred from the primary endosymbiont (cyanobacteria) to the primary host nucleus shortly after the primary endosymbiotic event or replaced with genes from other sources at an equally early time, i.e., before the formation of three primary plastid lineages. The one striking exception to this pattern is ferrochelatase, the enzyme catalyzing the first committed step to heme and bilin pigments. In this case, two red algal sequences do not cluster either with the other plastid sequences or with cyanobacterial sequences and appear to have a proteobacterial origin like that of the apicomplexan parasites Plasmodium and Toxoplasma. Although the heterokonts also acquired their plastid via secondary endosymbiosis from a red alga, the diatom has a typical plastid-cyanobacterial ferrochelatase. We have not found any remnants of the plastidlike heme pathway in the nonphotosynthetic heterokonts Phytophthora ramorum and Phytophthora sojae.

  11. A role for Haemophilus ducreyi Cu,ZnSOD in resistance to heme toxicity. (United States)

    Negari, Shahin; Sulpher, Jeff; Pacello, Francesca; Ingrey, Keely; Battistoni, Andrea; Lee, B Craig


    The Cu,Zn superoxide dismutase (Cu,ZnSOD) from Haemophilus ducreyi is the only enzyme of this class which binds a heme molecule at its dimer interface. To explore the role of the enzyme in this heme-obligate bacterium, a sodC mutant was created by insertional inactivation. No difference in growth rate was observed during heme limitation. In contrast, under heme rich conditions growth of the sodC mutant was impaired compared to the wild type strain. This growth defect was abolished by supplementation of exogenous catalase. Genetic complementation of the sodC mutant in trans demonstrated that the enzymatic property or the heme-binding activity of the protein could repair the growth defect of the sodC mutant. These results indicate that Cu,ZnSOD protects Haemophilus ducreyi from heme toxicity.

  12. Protective role of heme oxygenase-1 against inflammation in atherosclerosis. (United States)

    Durante, William


    Heme oxygenase-1 (HO-1) catalyzes the first and rate-limiting step in the metabolism of free heme into equimolar amounts of ferrous iron, carbon monoxide (CO), and biliverdin. Biliverdin is subsequently converted to bilirubin by biliverdin reductase. HO-1 has recently been identified as a promising therapeutic target in the treatment of vascular inflammatory disease, including atherosclerosis. HO-1 represses inflammation by removing the pro-inflammatory molecule heme and by generating CO and the bile pigments, biliverdin and bilirubin. These HO-1 reaction products are capable of blocking innate and adaptive immune responses by modifying the activation, differentiation, maturation, and/or polarization of numerous immune cells, including endothelial cells, monocytes/macrophages, dendritic cells, T lymphocytes, mast cells, and platelets. These cellular actions by CO and bile pigments result in diminished leukocyte recruitment and infiltration, and pro-inflammatory mediator production within atherosclerotic lesions. This review highlights the mechanisms by which HO-1 suppresses vascular inflammation in atherosclerosis, and explores possible therapeutic modalities by which HO-1 and its reaction products can be employed to ameliorate vascular inflammatory disease.

  13. Nitric oxide heme interactions in nitrophorin from Cimex lectularius (United States)

    Christmann, R.; Auerbach, H.; Berry, R. E.; Walker, F. A.; Schünemann, V.


    The nitrophorin from the bedbug Cimex lectularius (cNP) is a nitric oxide (NO) carrying protein. Like the nitrophorins (rNPs) from the kissing bug Rhodnius prolixus, cNP forms a stable heme Fe(III)-NO complex, where the NO can be stored reversibly for a long period of time. In both cases, the NPs are found in the salivary glands of blood-sucking bugs. The insects use the nitrophorins to transport the NO to the victim's tissues, resulting in vasodilation and reduced blood coagulation. However, the structure of cNP is significantly different to those of the rNPs from Rhodnius prolixus. Furthermore, the cNP can bind a second NO molecule to the proximal heme cysteine when present at higher concentrations. High field Mössbauer spectroscopy on 57Fe enriched cNP complexed with NO shows reduction of the heme iron and formation of a ferrous nitric oxide (Fe(II)-NO) complex. Density functional theory calculations reproduce the experimental Mössbauer parameters and confirm this observation.

  14. Facile synthesis of large-scale Ag nanosheet-assembled films with sub-10 nm gaps as highly active and homogeneous SERS substrates (United States)

    Li, Zhongbo; Meng, Guowen; Liang, Ting; Zhang, Zhuo; Zhu, Xiaoguang


    We report a facile low-cost synthetic approach to large-scale Ag nanosheet-assembled films with a high density of uniformly distributed sub-10 nm gaps between the adjacent nanosheets on Si substrates via galvanic cell reactions. The distribution density of Ag nanosheets on substrates could be tailored by tuning the duration of the HF-etching and the concentration of citric acid in the solution. Furthermore, in conjunction with a conventional photolithography, highly uniform patterned Ag nanosheet-assembled structures with different morphologies can be achieved on Si substrates via galvanic-cell-induced growth. By using rhodamine 6G as a standard test molecule, the large-scale Ag nanosheet-assembled films exhibit highly active and homogenous surface-enhanced Raman scattering (SERS) effect and also show promising potentials as reliable SERS substrates for rapid detection of trace polychlorinated biphenyls (PCBs).

  15. Synthesis and characterization of supramolecule self-assembly polyami-doamine (PAMAM G1-G1 NH2, CO2H end group Megamer

    Directory of Open Access Journals (Sweden)

    Omid Louie


    Full Text Available Supramolecule self-assembly polyamidoamine (PAMAM dendrimer refers to the chemical sys-tems made up of a discrete number of assembled molecular subunits or components. These strat-egies involve the covalent assembly of hierarchical components reactive monomers, branch cells or dendrons around atomic or molecular cores according to divergent/convergent dendritic branching principles, systematic filling of space around a core with shells (layers of branch cells. The polydispersity index (PDI for the supramolecule megamer are pretty closed to one, are in agreement with the Poisson probability distribution. Polyamidoamine (PAMAM den-drimer G1-G1 that it was PAMAM Megamer NH2, COOH end groupsynthesized and character-ized by FT-IR, 1H NMR, 13C NMRspectra and GelPermeation Chromatography (GPC.

  16. Heme metabolism in stress regulation and protein production: from Cinderella to a key player

    DEFF Research Database (Denmark)

    Martínez, J. L.; Petranovic, D.; Nielsen, Jens


    . Based on our recent findings and other recent reports, we here illustrate that heme is more than a co-factor. We also discuss the necessity to gain more insight into the heme biosynthesis pathway regulation, as this interacts closely with overall stress control. Understanding heme biosynthesis and its...... regulation could impact our ability to develop more efficient yeast cell factories for heterologous protein production....

  17. Porphyrin-induced photodynamic cross-linking of hepatic heme-binding proteins. (United States)

    Vincent, S H; Holeman, B; Cully, B C; Muller-Eberhard, U


    Three types of hepatic proteins, a heme-binding Z protein, a mixture of the glutathione S-transferases and a cytochrome P450 isozyme, were shown to be susceptible to photodynamic cross-linking and loss in antigenicity by naturally occurring porphyrins. At 50 microM, uroporphyrin caused the most and protoporphyrin the least photodecomposition. Hemopexin, a specific serum heme carrier, was photodecomposed but no cross-linking was detected. Heme and scavengers of singlet oxygen partially prevented protein photodecomposition.

  18. Construction of diverse supramolecular assemblies of dimetal subunits differing in coordinated water molecules via strong hydrogen bonding interactions: Synthesis, crystal structures and spectroscopic properties

    Indian Academy of Sciences (India)

    Sadhika Khullar; Sanjay K Mandal


    Three new supramolecular assemblies (constructed through strong hydrogen bonding) of [Co2(bpta)2(adc)(H2O)4](ClO4)2.2H2O (1), [Cu2(bpta)2(fum)(H2O)2](ClO4)2 (2) and [Cu2(bpta)2(tdc)(H2O) (ClO4)](ClO4).3H2O (3), which are synthesised by one pot self-assembly of the metal salt, bpta ligand and the corresponding dicarboxylate under the same reaction conditions, are reported (where adc = acetylene dicarboxylate, fum = fumarate, tdc = 2,5-thiophenedicarboxylate and bpta = N,N'-bis(2-pyridylmethyl)-tertbutylamine). These compounds have varying degrees of coordinatedwater molecules per dimetal subunits (four for 1, two for 2 and one for 3, respectively). Furthermore, the orientation of the coordinated water molecules in 1 and 2, with respect to the mono (carboxylato)-bridged dimetal subunit, is different (cis and trans, respectively). On the other hand, there is a coordinated perchlorate ion in 3 making the two metal centers inequivalent. Unlike 1 and 3, there are no lattice water molecules in 2. This difference in the dimetal subunit in 1-3 and the presence or absence of the lattice water molecules are the keys to forming the diverse supramolecular assemblies. In 1 and 3, the involvement of lattice water molecules in the construction of such assemblies is distinctly different. In case of 2, the formation of supramolecular assembly depends on the coordinated water molecule (trans to each other) and thus a ladder shaped supramolecular assembly is the result. The strength of hydrogen bonding observed in the networks of 1-3 is indicated in the O…O distances (2.596 Å to 3.160 Å) and the OH…O angles 124° to 176°. All are characterised by elemental analysis, FTIR spectroscopy and single crystal X-ray diffraction studies.

  19. Hydrothermal synthesis of one-dimensional assemblies of Pt nanoparticles and their sensor application for simultaneous determination of dopamine and ascorbic acid

    Energy Technology Data Exchange (ETDEWEB)

    Wang Wei [Qingdao University of Science and Technology, Key Laboratory of Nanostructured Materials (China); Wang Qingxiang [Zhangzhou Normal University (China); Zhang Zhikun, E-mail: [Qingdao University of Science and Technology, Key Laboratory of Nanostructured Materials (China)


    One-dimensional assemblies of Pt nanoparticles (NPs) with the size range of 10-20 nm have been synthesized via a simple hydrothermal route using soluble starch as both template and reducing agent. The formation mechanism of the product was studied in details. The electrochemical behavior of dopamine (DA) and ascorbic acid (AA) on the prepared one-dimensionally assembled Pt NPs modified glassy carbon electrode were studied by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques and showed satisfactory results for the simultaneous determination of DA and AA by resolving the overlapping voltammetric responses of DA and AA into two voltammetric peaks.

  20. Irradiation of bovine meat: effect of heme-iron concentration.; Irradiacao de carne bovina: efeito na concentracao de ferro heme

    Energy Technology Data Exchange (ETDEWEB)

    Mistura, Liliana Perazzini Furtado


    The irradiation is often used, nowadays, for meat conservation and it is important to know how much this process interferes with the nutritional quality of the meat. In this study round cut meat, ground and steaks (from a local supermarket) was irradiated with doses of O; 1; 2; 3; 4; 5; 7,5 and 10 kGy (JS-7500 Nordium Inc -Canada) and the interference of irradiation and the process of food preparation on heme-iron (H Fe) content was determined. Half of the sample was kept raw and the other half was grilled in a pre-warmed oven at 250 deg C for 9 min and a controlled humidity of 70%. The chemical composition, the total iron (T Fe) (EM) and the heme iron concentration were determined (Hornsey,1956) and the sensorial quality evaluated. The average T Fe concentration of raw and ground , ground and grilled, raw steaks and grilled steak meat, on dry and degreased basis was 113 mug/g, 121 mug/g , 91 mug/g and 77 mug/g; and the H Fe concentration 105 mug/g (93% of T Fe) , 88 mug/g (73% of T Fe), 90 mug/g (99% of T Fe) and 52 mug/g (68% of T Fe) respectively. Data were evaluated by ANOVA with fixed effects and multiple comparisons. The irradiation neither altered the chemical composition nor the proportion of heme iron of meat. The preparation conditions (temperature, cooking time, environment humidity, meat presentation) of the sample interfered more with the heme iron content than the irradiation. With the sensorial analysis we verified that meats irradiated with doses of 3 kGy were better evaluated in softness and succulency attributes than the others. Meat submitted to irradiation doses up to 3 kGy were accepted by the specialists' panel. (author)

  1. Altered heme catabolism by heme oxygenase-1 caused by mutations in human NADPH cytochrome P450 reductase

    Energy Technology Data Exchange (ETDEWEB)

    Pandey, Amit V., E-mail: [Pediatric Endocrinology, Diabetology and Metabolism, Department of Clinical Research, University of Bern, Tiefenaustrasse 120c, CH-3004 Bern (Switzerland); Flueck, Christa E.; Mullis, Primus E. [Pediatric Endocrinology, Diabetology and Metabolism, Department of Clinical Research, University of Bern, Tiefenaustrasse 120c, CH-3004 Bern (Switzerland)


    Research highlights: {yields} Mutations in POR identified from patients lead to reduced HO-1 activities. {yields} POR mutation Y181D affecting FMN binding results in total loss of HO-1 activity. {yields} POR mutations A287P, C569Y and V608F, lost 50-70% activity. {yields} Mutations in FAD binding domain, R457H, Y459H and V492E lost all HO-1 activity. {yields} POR polymorphisms P228L, R316W, G413S, A503V and G504R have normal activity. -- Abstract: Human heme oxygenase-1 (HO-1) carries out heme catabolism supported by electrons supplied from the NADPH through NADPH P450 reductase (POR, CPR). Previously we have shown that mutations in human POR cause a rare form of congenital adrenal hyperplasia. In this study, we have evaluated the effects of mutations in POR on HO-1 activity. We used purified preparations of wild type and mutant human POR and in vitro reconstitution with purified HO-1 to measure heme degradation in a coupled assay using biliverdin reductase. Here we show that mutations in POR found in patients may reduce HO-1 activity, potentially influencing heme catabolism in individuals carrying mutant POR alleles. POR mutants Y181D, A457H, Y459H, V492E and R616X had total loss of HO-1 activity, while POR mutations A287P, C569Y and V608F lost 50-70% activity. The POR variants P228L, R316W and G413S, A503V and G504R identified as polymorphs had close to WT activity. Loss of HO-1 activity may result in increased oxidative neurotoxicity, anemia, growth retardation and iron deposition. Further examination of patients affected with POR deficiency will be required to assess the metabolic effects of reduced HO-1 activity in affected individuals.

  2. A Novel Approach for Identifying the Heme-Binding Proteins from Mouse Tissues

    Institute of Scientific and Technical Information of China (English)

    Xiaolei Li; Rong Wang; Zhongsheng Sun; Zuyuan Xu; Jingyue Bao; Xiuqing Zhang; Xiaoli Feng; Siqi Liu; Xiaoshan Wang; Kang Zhao; Zhengfeng Zhou; Caifeng Zhao; Ren Yan; Liang Lin; Tingting Lei; Jianning Yin


    Heme is a key cofactor in aerobic life, both in eukaryotes and prokaryotes. Because of the high reactivity of ferrous protoporphyrin IX, the reactions of heme in cells are often carried out through heme-protein complexes. Traditionally studies of hemebinding proteins have been approached on a case by case basis, thus there is a limited global view of the distribution of heme-binding proteins in different cells or tissues. The procedure described here is aimed at profiling hemne-binding proteins in mouse tissues sequentially by 1) purification of heme-binding proteins by hemeagarose, an affinity chromatographic resin; 2) isolation of heme-binding proteins by SDS-PAGE or two-dimensional electrophoresis; 3) identification of heme-binding proteins by mass spectrometry. In five mouse tissues, over 600 protein spots were visualized on 2DE gel stained by Commassie blue and 154 proteins were identified by MALDI-TOF, in which most proteins belong to heme related. This methodology makes it possible to globally characterize the heme-binding proteins in a biological system.

  3. Nitrosylation of c heme in cd(1)-nitrite reductase is enhanced during catalysis. (United States)

    Rinaldo, Serena; Giardina, Giorgio; Cutruzzolà, Francesca


    The reduction of nitrite into nitric oxide (NO) in denitrifying bacteria is catalyzed by nitrite reductase. In several species, this enzyme is a heme-containing protein with one c heme and one d1 heme per monomer (cd1NiR), encoded by the nirS gene. For many years, the evidence of a link between NO and this hemeprotein represented a paradox, given that NO was known to tightly bind and, possibly, inhibit hemeproteins, including cd1NiRs. It is now established that, during catalysis, cd1NiRs diverge from "canonical" hemeproteins, since the product NO rapidly dissociates from the ferrous d1 heme, which, in turn, displays a peculiar "low" affinity for NO (KD=0.11 μM at pH 7.0). It has been also previously shown that the c heme reacts with NO at acidic pH but c heme nitrosylation was not extensively investigated, given that in cd1NiR it was considered a side reaction, rather than a genuine process controlling catalysis. The spectroscopic study of the reaction of cd1NiR and its semi-apo derivative (containing the sole c heme) with NO reported here shows that c heme nitrosylation is enhanced during catalysis; this evidence has been discussed in order to assess the potential of c heme nitrosylation as a regulatory process, as observed for cytochrome c nitrosylation in mammalian mitochondria.

  4. Natural chlorophyll but not chlorophyllin prevents heme-induced cytotoxic and hyperproliferative effects in rat colon. (United States)

    de Vogel, Johan; Jonker-Termont, Denise S M L; Katan, Martijn B; van der Meer, Roelof


    Diets high in red meat and low in green vegetables are associated with an increased risk of colon cancer. In rats, dietary heme, mimicking red meat, increases colonic cytotoxicity and proliferation of the colonocytes, whereas addition of chlorophyll from green vegetables inhibits these heme-induced effects. Chlorophyllin is a water-soluble hydrolysis product of chlorophyll that inhibits the toxicity of many planar aromatic compounds. The present study investigated whether chlorophyllins could inhibit the heme-induced luminal cytotoxicity and colonic hyperproliferation as natural chlorophyll does. Rats were fed a purified control diet, the control diet supplemented with heme, or a heme diet with 1.2 mmol/kg diet of chlorophyllin, copper chlorophyllin, or natural chlorophyll for 14 d (n = 8/group). The cytotoxicity of fecal water was determined with an erythrocyte bioassay and colonic epithelial cell proliferation was quantified in vivo by [methyl-(3)H]thymidine incorporation into newly synthesized DNA. Exfoliation of colonocytes was measured as the amount of rat DNA in feces using quantitative PCR analysis. Heme caused a >50-fold increase in the cytotoxicity of the fecal water, a nearly 100% increase in proliferation, and almost total inhibition of exfoliation of the colonocytes. Furthermore, the addition of heme increased TBARS in fecal water. Chlorophyll, but not the chlorophyllins, completely prevented these heme-induced effects. In conclusion, inhibition of the heme-induced colonic cytotoxicity and epithelial cell turnover is specific for natural chlorophyll and cannot be mimicked by water-soluble chlorophyllins.

  5. Benfang Lei’s research on heme acquisition in Gram-positive pathogens and bacterial pathogenesis

    Institute of Scientific and Technical Information of China (English)


    Benfang Lei’s laboratory conducts research on pathogenesis of human pathogen Group A Streptococcus (GAS)and horse pathogen Streptococcus equi(S.equi). His current research focuses on heme acquisition in Gram-positive pathogens and molecular mechanism of GAS and S.equi pathogenesis.Heme is an important source of essential iron for bacterial pathogens.Benfang Lei and colleagues identified the first cell surface heme-binding protein in Gram-positive pathogens and the heme acquisition system in GAS,demonstrated direct heme transfer from one protein to another,demonstrated an experimental pathway of heme acquisition by the Staphylococcus aureus Isd system,elucidated the activated heme transfer mechanism,and obtained evidence for a chemical mechanism of direct axial ligand displacement during the Shp-to-HtsA heme transfer reaction.These findings have considerably contributed to the progress that has been made over recent years in understanding the heme acquisition process in Grampositive pathogens.Pathogenesis of GAS is mediated by an abundance of extracellular proteins,and pathogenic role and functional mechanism are not known for many of these virulence factors.Lei laboratory identified a secreted protein of GAS as a CovRS-regulated virulence factor that is a protective antigen and is critical for GAS spreading in the skin and systemic dissemination.These studies may lead to development of novel strategies to prevent and treat GAS infections.

  6. In vivo and in vitro olefin cyclopropanation catalyzed by heme enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Coelho, Pedro S.; Brustad, Eric M.; Arnold, Frances H.; Wang, Zhan; Lewis, Jared C.


    The present invention provides methods for catalyzing the conversion of an olefin to any compound containing one or more cyclopropane functional groups using heme enzymes. In certain aspects, the present invention provides a method for producing a cyclopropanation product comprising providing an olefinic substrate, a diazo reagent, and a heme enzyme; and admixing the components in a reaction for a time sufficient to produce a cyclopropanation product. In other aspects, the present invention provides heme enzymes including variants and fragments thereof that are capable of carrying out in vivo and in vitro olefin cyclopropanation reactions. Expression vectors and host cells expressing the heme enzymes are also provided by the present invention.

  7. In vivo and in vitro olefin cyclopropanation catalyzed by heme enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Coelho, Pedro S; Brustad, Eric M; Arnold, Frances H; Wang, Zhan; Lewis, Jared C


    The present invention provides methods for catalyzing the conversion of an olefin to any compound containing one or more cyclopropane functional groups using heme enzymes. In certain aspects, the present invention provides a method for producing a cyclopropanation product comprising providing an olefinic substrate, a diazo reagent, and a heme enzyme; and admixing the components in a reaction for a time sufficient to produce a cyclopropanation product. In other aspects, the present invention provides heme enzymes including variants and fragments thereof that are capable of carrying out in vivo and in vitro olefin cyclopropanation reactions. Expression vectors and host cells expressing the heme enzymes are also provided by the present invention.

  8. TPR domain of NrfG mediates complex formation between heme lyase and formate-dependent nitrite reductase in Escherichia coli O157:H7. (United States)

    Han, Dohyun; Kim, Kyunggon; Oh, Jongkil; Park, Jungeun; Kim, Youngsoo


    Escherichia coli synthesize C-type cytochromes only during anaerobic growth in media supplemented with nitrate and nitrite. The reduction of nitrate to ammonium in the periplasm of Escherichia coli involves two separate periplasmic enzymes, nitrate reductase and nitrite reductase. The nitrite reductase involved, NrfA, contains cytochrome C and is synthesized coordinately with a membrane-associated cytochrome C, NrfB, during growth in the presence of nitrite or in limiting nitrate concentrations. The genes NrfE, NrfF, and NrfG are required for the formate-dependent nitrite reduction pathway, which involves at least two C-type cytochrome proteins, NrfA and NrfB. The NrfE, NrfF, and NrfG genes (heme lyase complex) are involved in the maturation of a special C-type cytochrome, apocytochrome C (apoNrfA), to cytochrome C (NrfA) by transferring a heme to the unusual heme binding motif of the Cys-Trp-Ser-Cys-Lys sequence in apoNrfA protein. Thus, in order to further investigate the roles of NrfG in the formation of heme lyase complex (NrfEFG) and in the interaction between heme lyase complex and formate-dependent nitrite reductase (NrfA), we determined the crystal structure of NrfG at 2.05 A. The structure of NrfG showed that the contact between heme lyase complex (NrfEFG) and NrfA is accomplished via a TPR domain in NrfG which serves as a binding site for the C-terminal motif of NrfA. The portion of NrfA that binds to TPR domain of NrfG has a unique secondary motif, a helix followed by about a six-residue C-terminal loop (the so called "hook conformation"). This study allows us to better understand the mechanism of special C-type cytochrome assembly during the maturation of formate-dependent nitrite reductase, and also adds a new TPR binding conformation to the list of TPR-mediated protein-protein interactions.

  9. Calcium-Dependent Conformation of a Heme and Fingerprint Peptide of the Di-Heme Cytochrome c Peroxidase from Paracoccus Pantotrophus

    Energy Technology Data Exchange (ETDEWEB)



    The structural changes in the heme macrocycle and substituents caused by binding of Ca{sup 2+} to the diheme cytochrome c peroxidase from Paracoccuspantotrophus were clarified by resonance Raman spectroscopy of the inactive filly oxidized form of the enzyme. The changes in the macrocycle vibrational modes are consistent with a Ca{sup 2+}-dependent increase in the out-of-plane distortion of the low-potential heme, the proposed peroxidatic heme. Most of the increase in out-of-plane distortion occurs when the high affinity site I is occupied, but a small further increase in distortion occurs when site II is also occupied by Ca{sup 2+}or Mg{sup 2+}. This increase in the heme distortion also explains the red shift in the Soret absorption band that occurs upon Ca{sup 2+} binding. Changes also occur in the low frequency substituent modes of the heme, indicating that a structural change in the covalently attached fingerprint pentapeptide of the LP heme occurs upon CM{sup 2+} binding to site I. These structural changes, possibly enhanced in the semi-reduced form of the enzyme, may lead to loss of the sixth ligand at the peroxidatic heme and activation of the enzyme.

  10. One-pot hydrothermal synthesis of hollow Fe3O4 microspheres assembled with nanoparticles for lithium-ion battery anodes

    DEFF Research Database (Denmark)

    Liu, Yanguo; Wang, Xiaoliang; Ma, Wuming


    Hollow Fe3O4 microspheres assembled with nanoparticles were successfully synthesized without the addition of any templates or subsequent treatments. When used as the anode materials for lithium-ion battery (LIB), the products showed good lithium storage properties, demonstrating their promising...

  11. Self-Assembled Dehydro[24]annulene Monolayers at the Liquid/Solid Interface: Toward On-Surface Synthesis of Tubular π-Conjugated Nanowires. (United States)

    Suzuki, Mitsuharu; Guo, Zhaoqi; Tahara, Kazukuni; Kotyk, Juliet F Khosrowabadi; Nguyen, Huan; Gotoda, Jun; Iritani, Kohei; Rubin, Yves; Tobe, Yoshito


    We have studied the self-assembly behavior of dehydro[24]annulene (D24A) derivatives 1, 2a-2d, and 3a-3c at the liquid/solid interface using scanning tunneling microscopy (STM). Both the relative placement and the nature of the four D24A substituents strongly influence the self-assembly pattern. Overall, the eight D24A derivatives examined in this study display seven types of 2D packing patterns. The D24A derivatives 1, 2a, and 3a have either two or four stearate groups and adopt face-on configurations of their macrocyclic cores with respect to the highly oriented pyrolytic graphite (HOPG) surface. Their 2D packing pattern is determined by the interchain spacings and number of stearate substituents. The D24A derivatives 2b-2d and 3b-3c bear hydrogen-bonding carbamate groups to further strengthen intermolecular interactions. Face-on patterns were also observed for most of these compounds, while an unstable edge-on self-assembly was observed in the case of 2b at room temperature. Stable edge-on self-assemblies of D24A derivatives were sought for this work as an important stepping stone to achieving the on-surface topochemical polymerization of these carbon-rich macrocycles into tubular π-conjugated nanowires. The overall factors determining the 2D packing patterns of D24As at the liquid/solid interface are discussed on the basis of theoretical simulations, providing useful guidelines for controlling the self-assembly pattern of future D24A macrocycles.

  12. O{sub 2}-mediated oxidation of ferrous nitrosylated human serum heme-albumin is limited by nitrogen monoxide dissociation

    Energy Technology Data Exchange (ETDEWEB)

    Ascenzi, Paolo, E-mail: [Interdepartmental Laboratory of Electron Microscopy, University Roma Tre, Via della Vasca Navale 79, I-00146 Roma (Italy); National Institute for Infectious Diseases I.R.C.C.S. ' Lazzaro Spallanzani' , Via Portuense 292, I-00149 Roma (Italy); Gullotta, Francesca; Gioia, Magda; Coletta, Massimo [Department of Experimental Medicine and Biochemical Sciences, University of Roma ' Tor Vergata' , Via Montpellier 1, I-00133 Roma (Italy); Interuniversity Consortium for the Research on the Chemistry of Metals in Biological Systems, Piazza Umberto I 1, I-87100 Bari (Italy); Fasano, Mauro [Department of Structural and Functional Biology, and Center of Neuroscience, University of Insubria, Via Alberto da Giussano 12a, I-21052 Busto Arsizio, VA (Italy)


    Research highlights: {yields} Human serum heme-albumin displays globin-like properties. {yields} O{sub 2}-mediated oxidation of ferrous nitrosylated human serum heme-albumin. {yields} Allosteric modulation of human serum heme-albumin reactivity. {yields} Rifampicin is an allosteric effector of human serum heme-albumin. {yields} Human serum heme-albumin is a ROS and NOS scavenger. -- Abstract: Human serum heme-albumin (HSA-heme-Fe) displays globin-like properties. Here, kinetics of O{sub 2}-mediated oxidation of ferrous nitrosylated HSA-heme-Fe (HSA-heme-Fe(II)-NO) is reported. Values of the first-order rate constants for O{sub 2}-mediated oxidation of HSA-heme-Fe(II)-NO (i.e., for ferric HSA-heme-Fe formation) and for NO dissociation from HSA-heme-Fe(II)-NO (i.e., for NO replacement by CO) are k = 9.8 x 10{sup -5} and 8.3 x 10{sup -4} s{sup -1}, and h = 1.3 x 10{sup -4} and 8.5 x 10{sup -4} s{sup -1}, in the absence and presence of rifampicin, respectively, at pH = 7.0 and T = 20.0 {sup o}C. The coincidence of values of k and h indicates that NO dissociation represents the rate limiting step of O{sub 2}-mediated oxidation of HSA-heme-Fe(II)-NO. Mixing HSA-heme-Fe(II)-NO with O{sub 2} does not lead to the formation of the transient adduct(s), but leads to the final ferric HSA-heme-Fe derivative. These results reflect the fast O{sub 2}-mediated oxidation of ferrous HSA-heme-Fe and highlight the role of drugs in modulating allosterically the heme-Fe-atom reactivity.

  13. Rapid induction of heme oxygenase 1 mRNA and protein by hyperthermia in rat brain: heme oxygenase 2 is not a heat shock protein.


    Ewing, J F; Maines, M D


    Catalytic activity of heme oxygenase (heme, hydrogen-donor:oxygen oxidoreductase, EC isozymes, HO-1 and HO-2, permits production of physiologic isomers of bile pigments. In turn, bile pigments biliverdin and bilirubin are effective antioxidants in biological systems. In the rat brain we have identified only the HO-1 isozyme of heme oxygenase as a heat shock protein and defined hyperthermia as a stimulus that causes an increase in brain HO-1 protein. Exposure of male rats to 42 degr...

  14. Reciprocal allosteric modulation of carbon monoxide and warfarin binding to ferrous human serum heme-albumin.

    Directory of Open Access Journals (Sweden)

    Alessio Bocedi

    Full Text Available Human serum albumin (HSA, the most abundant protein in human plasma, could be considered as a prototypic monomeric allosteric protein, since the ligand-dependent conformational adaptability of HSA spreads beyond the immediate proximity of the binding site(s. As a matter of fact, HSA is a major transport protein in the bloodstream and the regulation of the functional allosteric interrelationships between the different binding sites represents a fundamental information for the knowledge of its transport function. Here, kinetics and thermodynamics of the allosteric modulation: (i of carbon monoxide (CO binding to ferrous human serum heme-albumin (HSA-heme-Fe(II by warfarin (WF, and (ii of WF binding to HSA-heme-Fe(II by CO are reported. All data were obtained at pH 7.0 and 25°C. Kinetics of CO and WF binding to the FA1 and FA7 sites of HSA-heme-Fe(II, respectively, follows a multi-exponential behavior (with the same relative percentage for the two ligands. This can be accounted for by the existence of multiple conformations and/or heme-protein axial coordination forms of HSA-heme-Fe(II. The HSA-heme-Fe(II populations have been characterized by resonance Raman spectroscopy, indicating the coexistence of different species characterized by four-, five- and six-coordination of the heme-Fe atom. As a whole, these results suggest that: (i upon CO binding a conformational change of HSA-heme-Fe(II takes place (likely reflecting the displacement of an endogenous ligand by CO, and (ii CO and/or WF binding brings about a ligand-dependent variation of the HSA-heme-Fe(II population distribution of the various coordinating species. The detailed thermodynamic and kinetic analysis here reported allows a quantitative description of the mutual allosteric effect of CO and WF binding to HSA-heme-Fe(II.

  15. Cellular iron depletion weakens induction of heme oxygenase-1 by cadmium. (United States)

    Lai, Chengzhi; Loo, George


    Heme oxygenase-1 is an inducible cytoprotective gene, although its induction by environmental factors is not completely understood. This study aimed to ascertain if specific nutritive factors or related compounds influence heme oxygenase-1 expression. In HCT-116 cells, cadmium increased heme oxygenase-1 enzymatic activity. This effect of cadmium was weaker in cells made iron-deficient with the iron chelator, desferrioxamine, which was associated with repression of heme oxygenase-1 protein and mRNA expression. The repression by desferrioxamine of cadmium-induced heme oxygenase-1 upregulation was reversed upon iron replenishment of the cells. Additionally, it was found that thiol antioxidants inhibited the heme oxygenase-1 upregulation caused by cadmium and also by ethacrynic acid, which each decreased intracellular glutathione as did buthionine sulfoxamine. Interestingly, cadmium and ethacrynic acid increased nuclear translocation of Nrf2 and subsequent heme oxygenase-1 expression, but buthionine sulfoxamine did not. Furthermore, NADPH oxidase inhibitors (diphenyleneiodonium and apocynin, and a superoxide scavenger (Tiron) inhibited cadmium-induced upregulation of heme oxygenase-1. Diphenyleneiodonium was the most potent and inhibited NADPH-cytochrome P450 reductase as well, whereas apocynin and Tiron did not. It is concluded that adequate amounts of iron, which at the atomic level can serve as the pivotal element of heme in NADPH oxidase, must be present in cells to permit what appears to be thiol redox-sensitive, NADPH oxidase-dependent upregulation of heme oxygenase-1. Thus, these findings are significant because they suggest that cells without adequate iron would be unable to fully express the stress gene, heme oxygenase-1, when confronted with the toxic metal, cadmium.

  16. An expeditious synthesis of tailed tren-capped porphyrins. (United States)

    Even, Pascale; Ruzié, Christian; Ricard, David; Boitrel, Bernard


    [structure: see text] A one-pot two-step versatile synthesis of tailed tren-capped porphyrins has been achieved. The two resulting ligands demonstrate that this expeditious method can be applied to various axial bases to obtain highly functionalized macromolecules attractive for heme modeling purposes. Dioxygen binding of the pyridine-tailed iron complex is reported as a direct application.

  17. An Ag(I) energetic metal-organic framework assembled with the energetic combination of furazan and tetrazole: synthesis, structure and energetic performance. (United States)

    Qu, Xiao-Ni; Zhang, Sheng; Wang, Bo-Zhou; Yang, Qi; Han, Jing; Wei, Qing; Xie, Gang; Chen, San-Ping


    A novel Ag(I) energetic MOF [Ag16(BTFOF)9]n·[2(NH4)]n () assembled with Ag(iI ions and a furazan derivative, 4,4'-oxybis[3,3'-(1H-5-tetrazol)]furazan (H2BTFOF) was successfully synthesized and structurally characterized, featuring a three-dimensional porous structure incorporating ammonium cations. The thermal stability and energetic properties were determined, revealing that the 3D energetic MOF had an outstanding insensitivity (IS > 40 J), an ultrahigh detonation pressure (P) of 65.29 GPa and a detonation velocity (D) of 11.81 km cm(-3). In addition, the self-accelerating decomposition temperature (TSADT) and the critical temperature of thermal explosion (Tb) are also discussed in detail. The finding exemplifies that the assembly strategy plays a decisive role in the density and energetic properties of MOF-based energetic materials.

  18. Near infrared (NIR) lanthanide emissive Langmuir-Blodgett monolayers formed using Nd(III) directed self-assembly synthesis of chiral amphiphilic ligands. (United States)

    Barry, Dawn E; Kitchen, Jonathan A; Albrecht, Martin; Faulkner, Stephen; Gunnlaugsson, Thorfinnur


    The incorporation of chiral amphiphilic lanthanide-directed self-assembled Nd(III) complexes (Nd.13 and Nd.23) into stable Langmuir monolayers, and the subsequent Langmuir-Blodgett film formation of these, is described. The photophysical properties of the enantiomeric pair of ligands 1 and 2 in the presence of Nd(CF3SO3)3 were also investigated in CH3CN solutions using UV-vis, fluorescence, and lanthanide luminescence spectroscopies. Analysis of the resulting self-assembly processes revealed that two main species were formed in solution,1:1 and 1:3 Nd:L self-assembly complexes, with the latter being the dominant species upon the addition of 0.33 equivalents of Nd(III). Excited state lifetime measurements of Nd.13 and Nd.23 in CH3OH and CD3OD and CH3CN were also evaluated. The formation of the self-assembly in solution was also monitored by observing the changes in the circular dichroism (CD) spectra; and large differences were observed between the 1:3 and other stoichiometries in the spectra, allowing for correlation to be made with that seen in the emission studies of these systems. Surface pressure-area and surface pressure-time isotherms evidenced the formation of stable Langmuir monolayers of Nd.13 and Nd.23 at an air-water interface, and the deposition of these monolayers onto a quartz solid substrate (Langmuir-Blodgett films) gave rise to immobilized chiral monomolecular films which exhibited Nd(III) NIR luminescence upon excitation of the ligand chromophore, demonstrating efficient energy transfer to the Nd(III) excided state (sensitized) with concomitant emission centered at 800 and 1334 nm.

  19. Symposium DD: Low-Dimensional Materials-Synthesis, Assembly, Property Scaling and Modeling. Held in San Francisco, CA on April 9-13, 2007 (United States)


    quantum dots are exquisitely sensitive to extra charges. As for electrochromic materials, the optical absorption is directly modified by state filling...University, Daejeon, South Korea. Layer-by-layer( LbL ) self-assembly is a template-assisted process in which charge reversal techniques using...allows consecutive thin film deposition to proceed via the electrostatic LbL method. The persistence length is a basic property quantifying the stiffness

  20. Synthesis of a New Porphyrin-fluorescein Hybrid and its Supramolecular Self-assembly with Amino-porphyrinatomanganese(Ⅲ)by Hydrogen-bonding

    Institute of Scientific and Technical Information of China (English)

    Jia Zheng LU; Jin Wang HUANG; Li Fen FAN; Jie LIU; Ke Zhuan XU; Xian Li CHEN; Liang Nian JI


    A new porphyrin-fluorescein hybrid 2 (Fl-PPTPP) has been synthesized and characterized by UV-Vis, IR, 1H-NMR, ESI-MS and elemental analysis. The supramolecular self-assembly of Fl-PPTPP with amino-porphyrinatomanganese(Ⅲ) [Mn(Ⅲ) (p-APTPP)C1] by hydrogen-bonding was studied using fluorescence spectroscopic titration and ESI-MS.

  1. Sol-gel synthesis of mesoporous assembly of Nd 2O 3 nanocrystals with the aid of structure-directing surfactant (United States)

    Sreethawong, Thammanoon; Chavadej, Sumaeth; Ngamsinlapasathian, Supachai; Yoshikawa, Susumu


    The aim of this work was to demonstrate a simple route to synthesize mesoporous assembly of nanocrystalline neodymium oxide (Nd 2O 3) with predominant pore size distribution in the mesopore region, which was successfully done for the first time under mild conditions by a modified sol-gel process with the aid of structure-directing surfactant. Properly manipulated hydrolysis and condensation steps of neodymium n-butoxide modified with acetylacetone in the presence of laurylamine hydrochloride surfactant aqueous solution were performed to obtain the assembled Nd 2O 3 nanocrystal. Various techniques, including thermogravimetric and differential thermal analyses (TG-DTA), X-ray diffraction (XRD), N 2 adsorption-desorption, Brunauer-Emmett-Teller (BET) surface area analysis, Barrett-Joyner-Halenda (BJH) pore size distribution analysis, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and selected-area electron diffraction (SAED), were used to characterize the synthesized Nd 2O 3 nanocrystal. The N 2 adsorption-desorption isotherm of the synthesized Nd 2O 3 nanocrystal calcined at 500 °C exhibited single hysteresis loop with IUPAC type IV-like pattern, indicating the existence of foremost mesopore. BJH pore size distribution result revealed that the synthesized Nd 2O 3 nanocrystals possessed mostly mesopore due to their assembly with a small extent of macropore, resulting in very small mean pore diameter in the mesopore region. The particle size determined from both SEM and TEM micrographs was also extremely small of approximately 10 nm.

  2. Pilot-scale tests of HEME and HEPA dissolution process

    Energy Technology Data Exchange (ETDEWEB)

    Qureshi, Z.H.; Strege, D.K.


    A series of pilot-scale demonstration tests for the dissolution of High Efficiency Mist Eliminators (HEME`s) and High Efficiency Particulate Airfilters (HEPA) were performed on a 1/5th linear scale. These fiberglass filters are to be used in the Defense Waste Processing Facility (DWPF) to decontaminate the effluents from the off-gases generated during the feed preparation process and vitrification. When removed, these filters will be dissolved in the Decontamination Waste Treatment Tank (DWTT) using 5 wt% NaOH solution. The contaminated fiberglass is converted to an aqueous stream which will be transferred to the waste tanks. The filter metal structure will be rinsed with process water before its disposal as low-level solid waste. The pilot-scale study reported here successfully demonstrated a simple one step process using 5 wt% NaOH solution. The proposed process requires the installation of a new water spray ring with 30 nozzles. In addition to the reduced waste generated, the total process time is reduced to 48 hours only (66% saving in time). The pilot-scale tests clearly demonstrated that the dissolution process of HEMEs has two stages - chemical digestion of the filter and mechanical erosion of the digested filter. The digestion is achieved by a boiling 5 wt% caustic solutions, whereas the mechanical break down of the digested filter is successfully achieved by spraying process water on the digested filter. An alternate method of breaking down the digested filter by increased air sparging of the solution was found to be marginally successful are best. The pilot-scale tests also demonstrated that the products of dissolution are easily pumpable by a centrifugal pump.

  3. Formal synthesis of (+)-discodermolide. (United States)

    Francavilla, Charles; Chen, Weichun; Kinder, Frederick R


    [structure: see text] Herein we report the formal total synthesis of (+)-discodermolide in 21 steps (longest linear sequence) from commercially available Roche ester. This synthesis features the assembly of C(9-18) and C(19-24) fragments via a metal-chelated aldol coupling reaction.

  4. EPR of Mononuclear Non-Heme Iron Proteins


    Gaffney, Betty J.


    Flexible geometry of three- to six-protein side-chain ligands to non-heme iron in proteins is the basis for widely diverse reactivites ranging from iron transport to redox chemistry. The gap between fixed states determined by x-ray analysis can be filled by spectroscopic study of trapped intermediates. EPR is a versatile and relatively quick approach to defining intermediate states in terms of the geometry and electronic structures of iron. A number of examples in which the iron chemistry of ...

  5. Long-term dietary heme iron and red meat intake in relation to endometrial cancer risk

    NARCIS (Netherlands)

    Genkinger, J.M.; Friberg, E.; Goldbohm, R.A.; Wolk, A.


    Background: Heme and total iron, present in meat, have been hypothesized to promote carcinogenesis. Few prospective studies have examined the associations between intakes of heme and total iron, types of meat, and endometrial cancer risk. Objective: We evaluated the associations between intakes of h

  6. Detection, characterization, and screening of heme-binding molecules by mass spectrometry for malaria drug discovery

    NARCIS (Netherlands)

    Munoz-Durango, K.; Maciuk, A.; Harfouche, A.; Torijano-Gutierrez, S.; Jullian, J.C.; Quintin, J.; Spelman, K.; Mouray, E.; Grellier, P.; Figadere, B.


    Drug screening for antimalarials uses heme biocrystallization inhibition methods as an alternative to parasite cultures, but they involve complex processes and cannot detect artemisinin-like molecules. The described method detects heme-binding compounds by mass spectrometry, using dissociation of th

  7. Heme oxygenase-1 polymorphism is not associated with risk of colorectal cancer: a Danish prospective study

    DEFF Research Database (Denmark)

    Vogel, Ulla Birgitte; Andersen, Vibeke; Christensen, Jane;


    Objective: Intake of red and processed meat confers risk of colorectal cancer (CRC). We wanted to test whether heme in meat promotes carcinogenesis. Methods: Heme oxygenase-1 (HO-1, HMOX1) A-413T (rs2071746) was assessed in a nested case–cohort study of 383 CRC cases and 763 randomly selected...

  8. Natural chlorophyll but not chlorophyllin prevents heme-induced cytotoxic and hyperproliferative effects in rat colon

    NARCIS (Netherlands)

    Vogel, de J.; Jonker-Termont, D.S.M.L.; Katan, M.B.; Meer, van der R.


    Diets high in red meat and low in green vegetables are associated with an increased risk of colon cancer. In rats, dietary heme, mimicking red meat, increases colonic cytotoxicity and proliferation of the colonocytes, whereas addition of chlorophyll from green vegetables inhibits these heme-induced

  9. Effects of chemical modifications of heme on kinetics of carbon monoxide binding to free home

    Energy Technology Data Exchange (ETDEWEB)

    Sono, M.; McCray, J.A.; Asakura, T.


    The rates of carbon monoxide recombination to six different kinds of chemically modified heme with various substituents at positions 2 and 4 have been studied in the protein-free state (free heme) by the laser flash photolysis method in a mixture of ethylene glycol and 0.02 N NaOH (80:20, v/v) (80% ethylene glycol). The carbon monoxide combination rate constants to the various free hemes obtained in 80% ethylene glycol at 22/sup 0/ were 1.4, 2.1, 2.1, 3.7, 4.5, and 6.4 x 10/sup 7/ M/sup -1/ s/sup -1/ for 2,4-diformyl-, spirographis (2-formyl-4-vinyl-), isospirographis (2-vinyl-4-formyl-) proto-(2,4-divinyl-), deutero-(2,4-dihydrogen-), and meso-(2,4-diethyl-), hemes, respectively. This order of increase in carbon monoxide combination rate constants for these hemes correlates exactly with decrease in electron attractivity of heme side chains (i.e., increase in pK/sub 3/, basicity of nitrogen base of prophyrin) and is completely opposite to that obtained for carbon monoxide binding to these hemes reconstituted with apomyoglobin. Contrary to the results for myoglobin, the two isomers of monoformyl-monovinylheme exhibited similar optical properties and the same combination rate constant indicating that the differences in the optical and kinetic results observed in myoglobin are due to different interactions of these isomeric hemes with protein.

  10. Dietary heme adversely affects experimental colitis in rats, despite heat-shock protein induction

    NARCIS (Netherlands)

    Schepens, Marloes A. A.; Vink, Carolien; Schonewille, Arjan J.; Dijkstra, Gerard; van der Meer, Roelof; Bovee-Oudenhoven, Ingeborg M. J.


    Objective: Research on dietary modulation of inflammatory bowel disease is in its infancy. Dietary heme, mimicking red meat, is cytotoxic to colonic epithelium and thus may aggravate colitis. Alternatively, heme-induced colonic stress might also result in potential protective heat-shock proteins (HS

  11. Dynamic Motion and Rearranged Molecular Shape of Heme in Myoglobin: Structural and Functional Consequences

    Directory of Open Access Journals (Sweden)

    Saburo Neya


    Full Text Available Myoglobin, a simple oxygen binding protein, was reconstituted with various types of synthetic hemes to manipulate the heme-globin interactions. From the paramagnetic NMR analysis, small heme was found to rotate rapidly about the iron-histidine bond upon. This is a novel and typical example for the fluctuation of protein. The dynamic NMR analysis indicated that the 360° rotational rate of a small heme was 1,400 s−1 at room temperature. The X-ray analyses revealed that the tertiary structure of globin containing the smallest heme was closely similar to that of native protein despite extensive destruction of the specific heme-globin interactions. The functional analyses of O2 binding showed that the loose heme-globin contacts do not significantly affect the oxygen binding. On the other hand, the rearrangement of tetrapyrrole array and the non-planar deformation in porphyrin ring significantly affect the functional properties of myoglobin. These results, taken together, indicate that the essential factors to regulate the myoglobin function are hidden under the molecular shape of prosthetic group rather than in the nonbonded heme-globin contacts.

  12. Sabot assembly

    Energy Technology Data Exchange (ETDEWEB)

    Bzorgi, Fariborz


    A sabot assembly includes a projectile and a housing dimensioned and configured for receiving the projectile. An air pressure cavity having a cavity diameter is disposed between a front end and a rear end of the housing. Air intake nozzles are in fluid communication with the air pressure cavity and each has a nozzle diameter less than the cavity diameter. In operation, air flows through the plurality of air intake nozzles and into the air pressure cavity upon firing of the projectile from a gun barrel to pressurize the air pressure cavity for assisting in separation of the housing from the projectile upon the sabot assembly exiting the gun barrel.

  13. Crystal structure of the heme-IsdC complex, the central conduit of the Isd iron/heme uptake system in Staphylococcus aureus. (United States)

    Sharp, Katherine H; Schneider, Sabine; Cockayne, Alan; Paoli, Max


    Pathogens such as Staphylococcus aureus require iron to survive and have evolved specialized proteins to steal heme from their host. IsdC is the central conduit of the Isd (iron-regulated surface determinant) multicomponent heme uptake machinery; staphylococcal cell-surface proteins such as IsdA, IsdB, and IsdH are thought to funnel their molecular cargo to IsdC, which then mediates the transfer of the iron-containing nutrient to the membrane translocation system IsdDEF. The structure of the heme-IsdC complex reveals a novel heme site within an immunoglobulin-like domain and sheds light on its binding mechanism. The folding topology is reminiscent of the architecture of cytochrome f, cellobiose dehydrogenase, and ethylbenzene dehydrogenase; in these three proteins, the heme is bound in an equivalent position, but interestingly, IsdC features a distinct binding pocket with the ligand located next to the hydrophobic core of the beta-sandwich. The iron is coordinated with a tyrosine surrounded by several non-polar side chains that cluster into a tightly packed proximal side. On the other hand, the distal side is relatively exposed with a short helical peptide segment that acts as a lip clasping onto almost half of the porphyrin plane. This structural feature is argued to play a role in the mechanism of binding and release by switching to an open conformation and thus loosening the interactions holding the heme. The structure of the heme-IsdC complex provides a template for the understanding of other proteins, such as IsdA, IsdB, and IsdH, that contain the same heme-binding module as IsdC, known as the NEAT (near transporter) domain.

  14. Induced fit on heme binding to the Pseudomonas aeruginosa cytoplasmic protein (PhuS) drives interaction with heme oxygenase (HemO)



    Iron, an essential nutrient with limited bioavailability, requires specialized cellular mechanisms for uptake. Although iron uptake into the cytoplasm in the form of heme has been well characterized in many bacteria, the subsequent trafficking is poorly understood. The cytoplasmic heme-binding proteins belong to a structurally related family thought to have evolved as “induced fit” ligand-binding macromolecules. One member, Pseudomonas aeruginosa cytoplasmic protein (PhuS), has previously bee...

  15. Hemoglobin fructation promotes heme degradation through the generation of endogenous reactive oxygen species (United States)

    Goodarzi, M.; Moosavi-Movahedi, A. A.; Habibi-Rezaei, M.; Shourian, M.; Ghourchian, H.; Ahmad, F.; Farhadi, M.; Saboury, A. A.; Sheibani, N.


    Protein glycation is a cascade of nonenzymatic reactions between reducing sugars and amino groups of proteins. It is referred to as fructation when the reducing monosaccharide is fructose. Some potential mechanisms have been suggested for the generation of reactive oxygen species (ROS) by protein glycation reactions in the presence of glucose. In this state, glucose autoxidation, ketoamine, and oxidative advance glycation end products (AGEs) formation are considered as major sources of ROS and perhaps heme degradation during hemoglobin glycation. However, whether fructose mediated glycation produces ROS and heme degradation is unknown. Here we report that ROS (H2O2) production occurred during hemoglobin fructation in vitro using chemiluminescence methods. The enhanced heme exposure and degradation were determined using UV-Vis and fluorescence spectrophotometry. Following accumulation of ROS, heme degradation products were accumulated reaching a plateau along with the detected ROS. Thus, fructose may make a significant contribution to the production of ROS, glycation of proteins, and heme degradation during diabetes.

  16. Unsaturated Glycerophospholipids Mediate Heme Crystallization: Biological Implications for Hemozoin Formation in the Kissing Bug Rhodnius prolixus

    DEFF Research Database (Denmark)

    Stiebler, R.; Majerowicz, D.; Knudsen, Jens


    Hemozoin (Hz) is a heme crystal produced by some blood-feeding organisms, as an efficient way to detoxify heme derived from hemoglobin digestion. In the triatomine insect Rhodnius prolixus, Hz is essentially produced by midgut extracellular phospholipid membranes known as perimicrovillar membranes...... (PMVM). Here, we investigated the role of commercial glycerophospholipids containing serine, choline and ethanolamine as headgroups and R. prolixus midgut lipids (RML) in heme crystallization. All commercial unsaturated forms of phospholipids, as well as RML, mediated fast and efficient beta...... induced by uPE, and the other largely represented by crystals with numerous sharp edges and tapered ends. Heme crystallization reactions induced by RML were efficient, with a heme to beta-hematin conversion rate higher than 70%, but clearly slower (t1/2 of 9.9-17.7 minutes) than those induced by uPC and u...

  17. Hemoglobin fructation promotes heme degradation through the generation of endogenous reactive oxygen species. (United States)

    Goodarzi, M; Moosavi-Movahedi, A A; Habibi-Rezaei, M; Shourian, M; Ghourchian, H; Ahmad, F; Farhadi, M; Saboury, A A; Sheibani, N


    Protein glycation is a cascade of nonenzymatic reactions between reducing sugars and amino groups of proteins. It is referred to as fructation when the reducing monosaccharide is fructose. Some potential mechanisms have been suggested for the generation of reactive oxygen species (ROS) by protein glycation reactions in the presence of glucose. In this state, glucose autoxidation, ketoamine, and oxidative advance glycation end products (AGEs) formation are considered as major sources of ROS and perhaps heme degradation during hemoglobin glycation. However, whether fructose mediated glycation produces ROS and heme degradation is unknown. Here we report that ROS (H2O2) production occurred during hemoglobin fructation in vitro using chemiluminescence methods. The enhanced heme exposure and degradation were determined using UV-Vis and fluorescence spectrophotometry. Following accumulation of ROS, heme degradation products were accumulated reaching a plateau along with the detected ROS. Thus, fructose may make a significant contribution to the production of ROS, glycation of proteins, and heme degradation during diabetes.

  18. Cyanide does more to inhibit heme enzymes, than merely serving as an active-site ligand

    Energy Technology Data Exchange (ETDEWEB)

    Parashar, Abhinav [Center for Biomedical Research, VIT University, Vellore, Tamil Nadu, 632014 India (India); Venkatachalam, Avanthika [REDOx Lab, PSG Institute of Advanced Studies, Avinashi Road, Peelamedu, Coimbatore, Tamil Nadu, 641004 (India); Gideon, Daniel Andrew [Center for Biomedical Research, VIT University, Vellore, Tamil Nadu, 632014 India (India); Manoj, Kelath Murali, E-mail: [REDOx Lab, PSG Institute of Advanced Studies, Avinashi Road, Peelamedu, Coimbatore, Tamil Nadu, 641004 (India)


    Highlights: • Cyanide (CN) is a well-studied toxic principle, known to inhibit heme-enzymes. • Inhibition is supposed to result from CN binding at the active site as a ligand. • Diverse heme enzymes’ CN inhibition profiles challenge prevailing mechanism. • Poor binding efficiency of CN at low enzyme concentrations and ligand pressures. • CN-based diffusible radicals cause ‘non-productive electron transfers’ (inhibition). - Abstract: The toxicity of cyanide is hitherto attributed to its ability to bind to heme proteins’ active site and thereby inhibit their activity. It is shown herein that the long-held interpretation is inadequate to explain several observations in heme-enzyme reaction systems. Generation of cyanide-based diffusible radicals in heme-enzyme reaction milieu could shunt electron transfers (by non-active site processes), and thus be detrimental to the efficiency of oxidative outcomes.

  19. Thiol redox requirements and substrate specificities of recombinant cytochrome c assembly systems II and III. (United States)

    Richard-Fogal, Cynthia L; San Francisco, Brian; Frawley, Elaine R; Kranz, Robert G


    The reconstitution of biosynthetic pathways from heterologous hosts can help define the minimal genetic requirements for pathway function and facilitate detailed mechanistic studies. Each of the three pathways for the assembly of cytochrome c in nature (called systems I, II, and III) has been shown to function recombinantly in Escherichia coli, covalently attaching heme to the cysteine residues of a CXXCH motif of a c-type cytochrome. However, recombinant systems I (CcmABCDEFGH) and II (CcsBA) function in the E. coli periplasm, while recombinant system III (CCHL) attaches heme to its cognate receptor in the cytoplasm of E. coli, which makes direct comparisons between the three systems difficult. Here we show that the human CCHL (with a secretion signal) attaches heme to the human cytochrome c (with a signal sequence) in the E. coli periplasm, which is bioenergetically (p-side) analogous to the mitochondrial intermembrane space. The human CCHL is specific for the human cytochrome c, whereas recombinant system II can attach heme to multiple non-cognate c-type cytochromes (possessing the CXXCH motif.) We also show that the recombinant periplasmic systems II and III use components of the natural E. coli periplasmic DsbC/DsbD thiol-reduction pathway. This article is part of a Special Issue entitled: Biogenesis/Assembly of Respiratory Enzyme Complexes.

  20. Duodenal Absorption and Tissue Utilization of Dietary Heme and Nonheme Iron Differ in Rats123 (United States)

    Cao, Chang; Thomas, Carrie E.; Insogna, Karl L.; O'Brien, Kimberly O.


    Background: Dietary heme contributes to iron intake, yet regulation of heme absorption and tissue utilization of absorbed heme remains undefined. Objectives: In a rat model of iron overload, we used stable iron isotopes to examine heme- and nonheme-iron absorption in relation to liver hepcidin and to compare relative utilization of absorbed heme and nonheme iron by erythroid (RBC) and iron storage tissues (liver and spleen). Methods: Twelve male Sprague-Dawley rats were randomly assigned to groups for injections of either saline or iron dextran (16 or 48 mg Fe over 2 wk). After iron loading, rats were administered oral stable iron in the forms of 57Fe-ferrous sulfate and 58Fe-labeled hemoglobin. Expression of liver hepcidin and duodenal iron transporters and tissue stable iron enrichment was determined 10 d postdosing. Results: High iron loading increased hepatic hepcidin by 3-fold and reduced duodenal expression of divalent metal transporter 1 (DMT1) by 76%. Nonheme-iron absorption was 2.5 times higher than heme-iron absorption (P = 0.0008). Absorption of both forms of iron was inversely correlated with hepatic hepcidin expression (heme-iron absorption: r = −0.77, P = 0.003; nonheme-iron absorption: r = −0.80, P = 0.002), but hepcidin had a stronger impact on nonheme-iron absorption (P = 0.04). Significantly more 57Fe was recovered in RBCs (P = 0.02), and more 58Fe was recovered in the spleen (P = 0.01). Conclusions: Elevated hepcidin significantly decreased heme- and nonheme-iron absorption but had a greater impact on nonheme-iron absorption. Differential tissue utilization of heme vs. nonheme iron was evident between erythroid and iron storage tissues, suggesting that some heme may be exported into the circulation in a form different from that of nonheme iron. PMID:25332470


    Institute of Scientific and Technical Information of China (English)

    Guan-qun Zhong; Qing-hua Duan; Kui-lin Deng; Ai-qin Zhang; Ping Xie; Hai-liang Zhang; Rong-ben Zhang


    A novel soluble and reactive amide-bridged ladderlike polyhydrosiloxane (ALPHS) was first synthesized by an amido H-bonding self-assembled template. ALPHS with molecular weight M-n = 18300 has very highly ordered ladderlike structure, which was confirmed by a sharp resonance absorption peak of [-Si(H)O2/2 ] moiety with the half peak width A1/2 < 0.5 in 29Si-NMR spectrum. Presence of the reactive Si-H groups gives ALPHS an opportunity to further derive a variety of functional polymers by versatile Si-H reactions such as hydrosilylation, condensation, and so on.

  2. Self-Assembled Templates of Aromatic Pentapeptides for Synthesis of CdS Quantum-Dots to Detect the Trace Amounts of Hg(2+) in Aqueous Solutions. (United States)

    Meng, Min; Dou, Yingying; Xu, Wenlong; Hao, Jingcheng


    Molecular self-assembly has become a popular tool to prepare nanomaterials with potential applications, such as ion-responsive detection of Hg(2+) in aqueous solutions. In this study, FFACD aromatic pentapeptides, whose N-terminuses were protected by carboxyl (Ac-FFACD) or a 9-fluorenylmethoxycarbonyl group (Fmoc-FFACD), were chosen as building blocks to produce nanostructures in solutions. Based on the preliminary determination of the critical aggregation concentration (CAC) of Ac-FFACD and Fmoc-FFACD aromatic pentapeptides in water, the order of magnitude of which is 10(-5) mol·L(-1), self-assembled spiral and networked nanowires can be easily obtained over a range of concentrations. These nanowires were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), and atomic force microscopy (AFM). The self-assembled spiral and networked nanowires were designed to be used as templates for preparing CdS quantum dots (QDs) in-situ at room temperature. The peptide-functionalized, nanowire-encapsulated CdS QDs can be used for rapid, sensitive, and selective detection of trace amounts of mercuric ions (Hg(2+)) in aqueous solutions. This method enables rapid, linear detection (the linear correlation coefficients are 0.9972 of ΔF = 257.09 + 3.58 cHg(2+) for Ac-FFACD and 0.9994 of ΔF = 48.13 + 32.96 cHg(2+) for Fmoc-FFACD) with the Hg(2+) limit of detection at 300.85 ng·L(-1) and 32.09 ng·L(-1) for Ac-FFACD and Fmoc-FFACD, respectively. The supramolecular, self-assembled nanowires, fabricated from the two aromatic pentapeptides and having encapsulated QDs, exhibit superior properties attributable to the large loading capacity and the coordination sites of these peptides with Hg(2+). These structures can serve as novel Hg(2+) sensors and have possible applications for detection of various targets in scientific and engineering systems.

  3. Heme oxygenase-1 comes back to endoplasmic reticulum

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hong Pyo [School of Biological Sciences, Ulsan University (Korea, Republic of); Pae, Hyun-Ock [Department of Immunology, Wonkwang University School of Medicine (Korea, Republic of); Back, Sung Hun; Chung, Su Wol [School of Biological Sciences, Ulsan University (Korea, Republic of); Woo, Je Moon [Department of Opthalmology, Ulasn University Hospital (Korea, Republic of); Son, Yong [Department of Anesthesiology and Pain Medicine, Wonkwang University School of Medicine (Korea, Republic of); Chung, Hun-Taeg, E-mail: [School of Biological Sciences, Ulsan University (Korea, Republic of)


    Research highlights: {yields} Although multiple compartmentalization of HO-1 has been documented, the functional implication of this enzyme at these subcellular organelles is only partially elucidated. {yields} HO-1 expression at ER is induced by a diverse set of conditions that cause ER stressors. {yields} CO may induce HO-1 expression in human ECs by activating Nrf2 through PERK phosphorylation in a positive-feedback manner. {yields} ER-residing HO-1 and its cytoprotective activity against ER stress is discussed. -- Abstract: Originally identified as a rate-limiting enzyme for heme catabolism, heme oxygenase-1 (HO-1) has expanded its roles in anti-inflammation, anti-apoptosis and anti-proliferation for the last decade. Regulation of protein activity by location is well appreciated. Even though multiple compartmentalization of HO-1 has been documented, the functional implication of this enzyme at these subcellular organelles is only partially elucidated. In this review we discuss the endoplasmic reticulum (ER)-residing HO-1 and its cytoprotective activity against ER stress.

  4. Endogenous Estrogen-Mediated Heme Oxygenase Regulation in Experimental Menopause

    Directory of Open Access Journals (Sweden)

    Anikó Pósa


    Full Text Available Estrogen deficiency is one of the main causes of age-associated diseases in the cardiovascular system. Female Wistar rats were divided into four experimental groups: pharmacologically ovariectomized, surgically ovariectomized, and 24-month-old intact aging animals were compared with a control group. The activity and expression of heme oxygenases (HO in the cardiac left ventricle, the concentrations of cardiac interleukin-6 (IL-6 and tumor necrosis factor-α (TNF-α, the myeloperoxidase (MPO activity in the cardiac left ventricle, and the effects of heme oxygenase blockade (by 24-hour and 1-hour pretreatment with tin-protoporphyrin IX, SnPP on the epinephrine and phentolamine-induced electrocardiogram ST segment changes in vivo were investigated. The cardiac HO activity and the expression of HO-1 and HO-2 were significantly decreased in the aged rats and after ovariectomy. Estrogen depletion was accompanied by significant increases in the expression of IL-6 and TNF-α. The aged and ovariectomized animals exhibited a significantly elevated MPO activity and a significant ST segment depression. After pretreatment with SnPP augmented ST segment changes were determined. These findings demonstrate that the sensitivity to cardiac ischemia in estrogen depletion models is associated with suppression of the activity and expression of the HO system and increases in the secretion of proinflammatory cytokines and biomarkers.

  5. Heme oxygenase-1 accelerates erastin-induced ferroptotic cell death. (United States)

    Kwon, Min-Young; Park, Eunhee; Lee, Seon-Jin; Chung, Su Wol


    The oncogenic RAS-selective lethal small molecule Erastin triggers a unique iron-dependent form of nonapoptotic cell death termed ferroptosis. Ferroptosis is dependent upon the production of intracellular iron-dependent reactive oxygen species (ROS), but not other metals. However, key regulators remain unknown. The heme oxygenase (HO) is a major intracellular source of iron. In this study, the role of heme oxygenase in Erastin-triggered ferroptotic cancer cell death has been investigated. Zinc protoporphyrin IX (ZnPP), a HO-1 inhibitor, prevented Erastin-triggered ferroptotic cancer cell death. Furthermore, Erastin induced the protein and mRNA levels of HO-1 in HT-1080 fibrosarcoma cells. HO-1+/+ and HO-1-/- fibroblast, HO-1 overexpression, and chycloheximide-treated experiments revealed that the expression of HO-1 has a decisive effects in Erastin-triggered cell death. Hemin and CO-releasing molecules (CORM) promote Erastin-induced ferroptotic cell death, not by biliverdin and bilirubin. In addition, hemin and CORM accelerate the HO-1 expression in the presence of Erastin and increase membranous lipid peroxidation. Thus, HO-1 is an essential enzyme for iron-dependent lipid peroxidation during ferroptotic cell death.

  6. Modulation of Antiviral Immunity by Heme Oxygenase-1. (United States)

    Espinoza, Janyra A; González, Pablo A; Kalergis, Alexis M


    Heme oxygenase-1 (HO-1) is a stress-inducible, anti-inflammatory, and cytoprotective enzyme expressed in most cell types in the organism. Under several stress stimuli, HO-1 expression and activity is up-regulated to catalyze the rate-limiting enzymatic step of heme degradation into carbon monoxide, free iron, and biliverdin. Besides its effects on cell metabolism, HO-1 is also capable of modulating host innate and adaptive immune responses in response to sepsis, transplantation, and autoimmunity, and preventing oxidative damage associated with inflammation. In addition, recent studies have reported that HO-1 can exert a significant antiviral activity against a wide variety of viruses, including HIV, hepatitis C virus, hepatitis B virus, enterovirus 71, influenza virus, respiratory syncytial virus, dengue virus, and Ebola virus, among others. Herein, we address the current understanding of the functional significance of HO-1 against a variety of viruses and its potential as a therapeutic strategy to prevent and control viral infections. Furthermore, we review the most important features of the immunoregulatory functions for this enzyme.

  7. Studies of multi-heme cytochromes from Geobacter sulfurreducens

    Energy Technology Data Exchange (ETDEWEB)

    Pokkuluri, P. Raj; Londer, Yuri, Y.; Orshonsky, Valerie; Orshonsky, Lisa; Duke, Norma; Schiffer, Marianne


    The Geobacteraceae family predominates in the reduction of uranium in subsurface environments. We are focusing on the model organism, Geobacter sulfurreducens; its genome contains a large number (>100) of cytochromes c that function in metal reduction pathways. Intensive functional genomics and physiological studies are in progress in Prof. Derek Lovley's laboratory, and the complete genome sequence of this organism has been determined by Methe et al. 2003. We are studying cytochromes from the c{sub 7} family that are required for the reduction of Fe(III). Previously, we expressed in E. coli (Londer et al., 2002) and determined the three-dimensional structure at 1.45 {angstrom} resolution (Pokkuluri et al., 2004a) of the three-heme cytochrome c{sub 7} (PpcA, coded by ORF01023) characterized by Lloyd et al., 2003. Further we identified in the G. sulfurreducens genome ORFs for several of its homologs (Pokkuluri et al., 2004a). Four of the ORFs are the same size as PpcA; three other ORFs are polymers of c7-type domains, two of which consist of four domains and one of nine domains, that contain 12 and 27 hemes respectively.

  8. Heme oxygenase-1 against vascular insufficiency: roles of atherosclerotic disorders. (United States)

    Ishikawa, Kazunobu


    Heme oxygenase (HO), an enzyme essential for heme degradation, shows anti-oxidative and anti-inflammatory properties via the production of bile pigments, carbon monoxide (CO) and ferritin induction under various pathophysiological conditions. A number of recent studies have shown biological effects of HO reaction in cardiovascular disorders. An inducible form of HO, HO-1, is induced by a variety of stresses such as oxidized lipoproteins, cytokines, hemodynamic changes, angiotensin II and nitric oxide (NO) in vascular wall. HO-1 induction seems to function as an adaptive response against these injurious stimuli. HO-1 induction in artery wall scavenges reactive oxygen species, which leads to the attenuation of monocyte adhesion and chemotaxis. HO-1 induction also reduces lipid peroxidation in plasma and artery wall. These properties of HO-1 suggest anti-atherogenic roles of this enzyme. In this review, roles of endothelial HO-1 expression and bilirubin in atherogenesis are also discussed. HO-1 also seems to play a significant role in restenosis after angioplasty, which is a major clinical problem associated with atherosclerosis. Recent progress in human HO-1 genetics supports these experimental results. This review aims to reaffirm current problems in the biological aspects of HO and suggest future research direction and clinical application.

  9. Heme and menaquinone induced electron transport in lactic acid bacteria

    Directory of Open Access Journals (Sweden)

    Santos Filipe


    Full Text Available Abstract Background For some lactic acid bacteria higher biomass production as a result of aerobic respiration has been reported upon supplementation with heme and menaquinone. In this report, we have studied a large number of species among lactic acid bacteria for the existence of this trait. Results Heme- (and menaquinone stimulated aerobic growth was observed for several species and genera of lactic acid bacteria. These include Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacilllus brevis, Lactobacillus paralimentarius, Streptococcus entericus and Lactococcus garviae. The increased biomass production without further acidification, which are respiration associated traits, are suitable for high-throughput screening as demonstrated by the screening of 8000 Lactococcus lactis insertion mutants. Respiration-negative insertion-mutants were found with noxA, bd-type cytochrome and menaquinol biosynthesis gene-disruptions. Phenotypic screening and in silico genome analysis suggest that respiration can be considered characteristic for certain species. Conclusion We propose that the cyd-genes were present in the common ancestor of lactic acid bacteria, and that multiple gene-loss events best explains the observed distribution of these genes among the species.

  10. Bidentate ligation of heme analogues; novel biomimetics of peroxidase active site. (United States)

    Ashkenasy, Gonen; Margulies, David; Felder, Clifford E; Shanzer, Abraham; Powers, Linda S


    The multifunctional nature of proteins that have iron-heme cofactors with noncovalent histidine linkage to the protein is controlled by the heme environment. Previous studies of these active-site structures show that the primary difference is the length of the iron-proximal histidine bond, which can be controlled by the degree of H-bonding to this histidine. Great efforts to mimic these functions with synthetic analogues have been made for more than two decades. The peroxidase models resulted in several catalytic systems capable of a large range of oxidative transformations. Most of these model systems modified the porphyrin ring covalently by directly binding auxiliary elements that control and facilitate reactivity; for example, electron-donating or -withdrawing substituents. A biomimetic approach to enzyme mimicking would have taken a different route, by attempting to keep the porphyrin ring system unaltered, as close as possible to its native form, and introducing all modifications at or close to the axial coordination sites. Such a model system would be less demanding synthetically, would make it easy to study the effect of a single structural modification, and might even provide a way to probe effects resulting from porphyrin exchange. We introduce here an alternative model system based on these principles. It consists of a two component system: a bis-imidazolyl ligand and an iron-porphyrin (readily substituted by a hemin). All modifications were introduced only to the ligand that engulfs the porphyrin and binds to the iron's fifth and sixth coordination sites. We describe the design, synthesis, and characterization of nine different model compounds with increased complexity. The primary tool for characterizing the environment of each complex Fe(III) center was the Extended X-ray Absorption Fine Structure (EXAFS) measurements, supported by UV/Vis, IR, and NMR spectroscopy and by molecular modeling. Introduction of asymmetry, by attaching different imidazoles

  11. Synthesis and self-assembly of dumbbell shaped ZnO sub-micron structures using low temperature chemical bath deposition technique

    Energy Technology Data Exchange (ETDEWEB)

    Borade, P. [National Centre for Nanoscience and Nanotechnology, University of Mumbai, Kalina Campus, Santacruz (E), Mumbai 400098 (India); Joshi, K.U. [Anton-Paar India Pvt. Ltd., Thane (W), 400607 (India); Gokarna, A.; Lerondel, G. [Laboratoire de Nanotechnologie et D' Instrumentation Optique, Institut Charles Delaunay, CNRS UMR 6281, Université de Technologie de Troyes, 12 Rue Marie Curie, BP 2060, 10010 Troyes (France); Walke, P. [National Centre for Nanoscience and Nanotechnology, University of Mumbai, Kalina Campus, Santacruz (E), Mumbai 400098 (India); Late, D. [National Chemical Laboratory (NCL), Pune 400027 (India); Jejurikar, S.M., E-mail: [National Centre for Nanoscience and Nanotechnology, University of Mumbai, Kalina Campus, Santacruz (E), Mumbai 400098 (India)


    We report well dispersed horizontal growth of ZnO sub-micron structures using simplest technique ever known i.e. chemical bath deposition (CBD). A set of samples were prepared under two different cases A) dumbbell shaped ZnO grown in CBD bath and B) tubular ZnO structures evolved from dumbbell shaped structures by dissolution mechanism. Single phase wurtzite ZnO formation is confirmed using X-ray diffraction (XRD) technique in both cases. From the morphological investigations performed using scanning electron microscopy (SEM), sample prepared under case A indicate formation of hex bit tool (HBT) shaped ZnO crystals, which observed to self-organize to form dumbbell structures. Further these microstructures are then converted into tubular structures as a fragment of post CBD process. The possible mechanism responsible for the self-assembly of HBT units to form dumbbell structures is discussed. Observed free excitonic peak located at 370 nm in photoluminescence (PL) spectra recorded at 18 K indicate that the micro/nanostructures synthesized using CBD are of high optical quality. - Highlights: • Controlled growth of Dumbbell shaped ZnO using Chemical Bath Deposition (CBD). • Growth mechanism of dumbbell shaped ZnO by self-assembling was discussed. • Quick Transformation of ZnO dumbbell structures in to tubular structures by dissolution. • Sharp UV Emission at 370 nm from both dumbbell and tubular structures.

  12. In Situ Electrochemical Synthesis and Deposition of Discotic Hexa-peri-hexabenzocoronene Molecules on Electrodes: Self-Assembled Structure, Redox Properties, and Application for Supercapacitor. (United States)

    Qin, Leiqiang; Zhang, Yunan; Wu, Xiaoyan; Nian, Li; Xie, Zengqi; Liu, Linlin; Ma, Yuguang


    Discotic hexa-peri-hexabenzocoronene (HBC) molecules are synthesized by electrochemical cyclodehydrogenation reaction and in situ self-assembled to π-electronic, discrete nanofibular objects with an average diameter about 70 nm, which are deposited directly onto the electrode. The nanofibers consist of columnar arrays of the π-stacked HBC molecules and the intercolumnar distance is determined to be 1.19 nm by X-ray diffraction, which corresponds well to the distance of 1.1 nm observed by high-resolution transmitting electron microscopy. The diameter of the molecular columns matches the size of the discotic HBC molecule indicating face-to-face π-stacking of HBC units in the column. The HBC nanofibers on electrode are redox active, and the nanosized columnar structures provide a huge surface area, which is a great benefit for the charging/discharging process, delivering excellent capacitance of 155 F g(-1) . The described electrochemical deposition method shows great advantage for self-assembling the family of insoluble and structurally designable graphene-like nano materials, which constitutes an important step toward molecular electronics.

  13. Synthesis of novel 3D SnO flower-like hierarchical architectures self-assembled by nano-leaves and its photocatalysis

    Energy Technology Data Exchange (ETDEWEB)

    Cui, Yongkui; Wang, Fengping, E-mail:; Iqbal, M. Zubair; Wang, Ziya; Li, Yan; Tu, Jianhai


    Highlights: • Novel 3D SnO flowers self-assembled by 2D nano-leaves were synthesized by hydrothermal method. • The SnO nano-leaf is of single crystalline nature. • The band gap of 2.59 eV of as-prepared products was obtained. • The as-synthesized material will be a promising photocatalytic material. - Abstract: In this report, the novel 3D SnO flower-like hierarchical architectures self-assembled by 2D SnO nano-leaves are successfully synthesized via template-free hydrothermal approach under facile conditions. The high-resolution transmission electron microscopy results demonstrate that the 2D nano-leaves structure is of single crystalline nature. The band gap 2.59 eV for prepared product is obtained from UV–vis diffuse reflectance spectrum. The photocatalysis of the as prepared SnO for degrading methyl orange (MO) has been studied. A good photocatalytic activity is obtained and the mechanism is discussed in detail. Results indicate that the SnO nanostructures are the potential candidates for photocatalyst applications.

  14. Solvothermal synthesis of size-tunable ZnFe2O4 colloidal nanocrystal assemblies and their electrocatalytic activity towards hydrogen peroxide (United States)

    Liu, Ruirui; Lv, Meng; Wang, Qianbin; Li, Hongliang; Guo, Peizhi; Zhao, X. S.


    Three ZnFe2O4 colloidal nanocrystal assemblies (CNAs), namely CNA1, CNA2 and CNA3, have been synthesized solvothermally with the size of 560 nm, 460 nm and 330 nm and are formed by the self-assembly of primary nanocrystals with the crystallite sizes of 19.2 nm, 15.5 nm and 21.8 nm, respectively. It was found that CNA2 performed superparamagnetic behavior with a saturation magnetization value of 36.9 emu g-1 while either CNA1 or CNA3 exhibited weak ferromagnetic with a small hysteresis loop and large saturation magnetization. Electrochemical sensing measurements toward the reduction of hydrogen peroxide showed that the peak currents of the CNAs in cyclic voltammograms showed a linear relationship with the concentration of hydrogen peroxide in the experimental conditions and the peak potentials were increased with the order of CNA3, CNA2 and CNA1. The formation mechanism of ZnFe2O4 CNAs had been discussed based on the experimental data. The magnetism and electrocatalysis of the ZnFe2O4 CNAs were supposed to be dependent on the size of primary nanoparticles and the structure of the CNAs.

  15. Synthesis of Upconverting Hydrogel Nanocomposites Using Thiol-Ene Click Chemistry: Template for the Formation of Dendrimer-Like Gold Nanoparticle Assemblies. (United States)

    Meesaragandla, Brahmaiah; Mahalingam, Venkataramanan


    The synthesis of upconverting hydrogel nanocomposites by base-catalyzed thiol-ene click reaction between 10-undecenoic acid capped Yb(3+)/Er(3+)-doped NaYF4 nanoparticles and pentaerythritol tetrakis(3-mercaptopropionate) (PETMP) as tetrathiol monomer is reported. This synthetic strategy for nanocomposite gels is quite different from works where usually the preformed gels are mixed with the nanoparticles. Developing nanocomposites by surface modification of capping ligands would allow tuning and controlling of the separation of the nanoparticles inside the gel network. The hydrogel nanocomposites prepared by thiol-ene click reaction show strong enhancement in luminescence intensity compared to 10-undecenoic acid-capped Yb(3+)/Er(3+)-doped NaYF4 nanoparticles through the upconversion process (under 980 nm laser excitation). The hydrogel nanocomposites display strong swelling characteristics in water resulting in porous structures. Interestingly, the resulting nanocomposite gels act as templates for the synthesis of dendrimer-like Au nanostructures when HAuCl4 is reduced in the presence of the nanocomposite gels.

  16. Dump assembly (United States)

    Goldmann, Louis H.


    A dump assembly having a fixed conduit and a rotatable conduit provided with overlapping plates, respectively, at their adjacent ends. The plates are formed with openings, respectively, normally offset from each other to block flow. The other end of the rotatable conduit is provided with means for securing the open end of a filled container thereto. Rotation of the rotatable conduit raises and inverts the container to empty the contents while concurrently aligning the conduit openings to permit flow of material therethrough.

  17. Bioactive dietary polyphenols decrease heme iron absorption by decreasing basolateral iron release in human intestinal Caco-2 cells. (United States)

    Ma, Qianyi; Kim, Eun-Young; Han, Okhee


    Because dietary polyphenolic compounds have a wide range of effects in vivo and vitro, including chelation of metals such as iron, it is prudent to test whether the regular consumption of dietary bioactive polyphenols impair the utilization of dietary iron. Because our previous study showed the inhibitory effect of (-) -epigallocatechin-3-gallate (EGCG) and grape seed extract (GSE) on nonheme iron absorption, we investigated whether EGCG and GSE also affect iron absorption from heme. The fully differentiated intestinal Caco-2 cells grown on microporous membrane inserts were incubated with heme (55)Fe in uptake buffer containing EGCG or GSE in the apical compartment for 7 h. Both EGCG and GSE decreased (P heme-derived iron. However, apical heme iron uptake was increased (P heme (55)Fe, the transfer of iron across the intestinal basolateral membrane was extremely low, indicating that basolateral export was impaired by GSE. In contrast, EGCG moderately decreased the cellular assimilation of heme (55)Fe, but the basolateral iron transfer was extremely low, suggesting that the basolateral efflux of heme iron was also inhibited by EGCG. Expression of heme oxygenase, ferroportin, and hephaestin protein was not changed by EGCG and GSE. The apical uptake of heme iron was temperature dependent and saturable in fully differentiated Caco-2 cells. Our data show that bioactive dietary polyphenols inhibit heme iron absorption mainly by reducing basolateral iron exit rather than decreasing apical heme iron uptake in intestinal cells.

  18. General Assembly

    CERN Multimedia

    Staff Association


    5th April, 2016 – Ordinary General Assembly of the Staff Association! In the first semester of each year, the Staff Association (SA) invites its members to attend and participate in the Ordinary General Assembly (OGA). This year the OGA will be held on Tuesday, April 5th 2016 from 11:00 to 12:00 in BE Auditorium, Meyrin (6-2-024). During the Ordinary General Assembly, the activity and financial reports of the SA are presented and submitted for approval to the members. This is the occasion to get a global view on the activities of the SA, its financial management, and an opportunity to express one’s opinion, including taking part in the votes. Other points are listed on the agenda, as proposed by the Staff Council. Who can vote? Only “ordinary” members (MPE) of the SA can vote. Associated members (MPA) of the SA and/or affiliated pensioners have a right to vote on those topics that are of direct interest to them. Who can give his/her opinion? The Ordinary General Asse...

  19. HRG1 is essential for heme transport from the phagolysosome of macrophages during erythrophagocytosis. (United States)

    White, Carine; Yuan, Xiaojing; Schmidt, Paul J; Bresciani, Erica; Samuel, Tamika K; Campagna, Dean; Hall, Caitlin; Bishop, Kevin; Calicchio, Monica L; Lapierre, Ariane; Ward, Diane M; Liu, Paul; Fleming, Mark D; Hamza, Iqbal


    Adult humans have about 25 trillion red blood cells (RBCs), and each second we recycle about 5 million RBCs by erythrophagocytosis (EP) in macrophages of the reticuloendothelial system. Despite the central role for EP in mammalian iron metabolism, the molecules and pathways responsible for heme trafficking during EP remain unknown. Here, we show that the mammalian homolog of HRG1, a transmembrane heme permease in C. elegans, is essential for macrophage iron homeostasis and transports heme from the phagolysosome to the cytoplasm during EP. HRG1 is strongly expressed in macrophages of the reticuloendothelial system and specifically localizes to the phagolysosomal membranes during EP. Depletion of Hrg1 in mouse macrophages causes attenuation of heme transport from the phagolysosomal compartment. Importantly, missense polymorphisms in human HRG1 are defective in heme transport. Our results reveal HRG1 as the long-sought heme transporter for heme-iron recycling in macrophages and suggest that genetic variations in HRG1 could be modifiers of human iron metabolism.

  20. Effects of heme iron enriched peptide on iron deficiency anemia in rats. (United States)

    Tang, Ning; Chen, Le-qun; Zhuang, Hong


    The present study aims to investigate whether a daily intake of heme iron enriched peptide obtained from bovine hemoglobin is effective in alleviating iron deficiency anemia (IDA). Wistar rats were randomly divided into six groups: a control group, an anemic group not treated, and anemic groups treated with FeSO4 or with the heme iron enriched peptide at low, moderate or high doses. The rats in the anemic groups were fed on a low-iron diet to establish the iron deficiency anemia model. After the model had been established, different doses of heme iron enriched peptide were given to the rats once a day via intragastric administration. After the iron supplement administration, it was observed that heme iron enriched peptide had effective restorative action returning the hemoglobin, red blood cells, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin concentration and serum iron in IDA animals to normal values or better. In addition, compared with FeSO4, higher Fe bioavailability and fewer side effects were observed. The rats in the moderate dose group had the highest apparent Fe absorption. Moreover, in vivo antioxidant activity was also observed, enhancing the activities of antioxidant enzymes and reduced malondialdehyde levels in IDA rats. Furthermore, the heme iron enriched peptide also exhibited strong in vitro antioxidant activities. In conclusion, heme iron enriched peptide significantly alleviated iron deficiency anemia, and exhibited strong in vitro and in vivo antioxidant activities. This suggests that heme iron enriched peptide might be exploited as a safe, efficient new iron supplement.

  1. Therapeutic Approaches to Limit Hemolysis-Driven Endothelial Dysfunction: Scavenging Free Heme to Preserve Vasculature Homeostasis

    Directory of Open Access Journals (Sweden)

    Francesca Vinchi


    Full Text Available Hemolysis results in the release of hemoglobin and heme into the bloodstream and is associated with the development of several pathologic conditions of different etiology, including hemoglobinopathies, hemolytic anemias, bacterial infections, malaria, and trauma. In addition, hemolysis is associated with surgical procedures, hemodialysis, blood transfusion, and other conditions in which mechanical forces can lead to red blood cell rupture. Free plasma hemoglobin and heme are toxic for the vascular endothelium since heme iron promotes oxidative stress that causes endothelial activation responsible for vasoocclusive events and thrombus formation. Moreover, free hemoglobin scavenges nitric oxide, reducing its bioavailability, and heme favours ROS production, thus causing oxidative nitric oxide consumption. This results in the dysregulation of the endothelium vasodilator:vasoconstrictor balance, leading to severe vasoconstriction and hypertension. Thus, endothelial dysfunction and impairment of cardiovascular function represent a common feature of pathologic conditions associated with hemolysis. In this review, we discuss how hemoglobin/heme released following hemolysis may affect vascular function and summarise the therapeutic approaches available to limit hemolysis-driven endothelial dysfunction. Particular emphasis is put on recent data showing the beneficial effects obtained through the use of the plasma heme scavenger hemopexin in counteracting heme-mediated endothelial damage in mouse models of hemolytic diseases.

  2. Elucidating second coordination sphere effects in heme proteins using low-temperature magnetic circular dichroism spectroscopy. (United States)

    Lehnert, Nicolai


    This paper reviews recent findings on how the second coordination sphere of heme proteins fine-tunes the properties of the heme active site via hydrogen bonding. This insight is obtained from low-temperature magnetic circular dichroism (MCD) spectroscopy. In the case of high-spin ferric hemes, MCD spectroscopy allows for the identification of a multitude of charge-transfer (CT) transitions. Using optically-detected magnetic saturation curves, out-of-plane polarized CT transitions between the heme and its axial ligand(s) can be identified. In the case of ferric Cytochrome P450cam, the corresponding S(σ)→Fe(III) CT transition can be used as a probe for the {Fe(III)-axial ligand} interaction, indicating that the hydrogen bonding network of the proximal Cys only plays a limited role for fine-tuning the Fe(III)-S(Cys) interaction. In the case of high-spin ferrous hemes with axial His/imidazole coordination, our MCD-spectroscopic investigations have uncovered a direct correlation between the strength of the hydrogen bond to the proximal imidazole ligand and the ground state of the complexes. With neutral imidazole coordination, the doubly occupied d-orbital of high-spin iron(II) is of d(π) character, located orthogonal to the heme plane. As the strength of the hydrogen bond increases, this orbital rotates into the heme plane, changing the ground state of the complex.

  3. Heme-induced Trypanosoma cruzi proliferation is mediated by CaM kinase II

    Energy Technology Data Exchange (ETDEWEB)

    Souza, C.F. [Laboratorio de Imunomodulacao e Protozoologia, Instituto Oswaldo Cruz, Fiocruz (Brazil); Carneiro, A.B.; Silveira, A.B. [Laboratorio de Sinalizacao Celular, Instituto de Bioquimica Medica, UFRJ (Brazil); Laranja, G.A.T. [Laboratorio de Interacao Tripanosomatideos e Vetores, Departamento de Bioquimica, IBRAG, UERJ, 20551-030 Rio de Janeiro (Brazil); Silva-Neto, M.A.C. [Laboratorio de Sinalizacao Celular, Instituto de Bioquimica Medica, UFRJ (Brazil); INCT, Entomologia Molecular (Brazil); Costa, S.C. Goncalves da [Laboratorio de Imunomodulacao e Protozoologia, Instituto Oswaldo Cruz, Fiocruz (Brazil); Paes, M.C., E-mail: [Laboratorio de Interacao Tripanosomatideos e Vetores, Departamento de Bioquimica, IBRAG, UERJ, 20551-030 Rio de Janeiro (Brazil); INCT, Entomologia Molecular (Brazil)


    Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted through triatomine vectors during their blood-meal on vertebrate hosts. These hematophagous insects usually ingest approximately 10 mM of heme bound to hemoglobin in a single meal. Blood forms of the parasite are transformed into epimastigotes in the crop which initiates a few hours after parasite ingestion. In a previous work, we investigated the role of heme in parasite cell proliferation and showed that the addition of heme significantly increased parasite proliferation in a dose-dependent manner . To investigate whether the heme effect is mediated by protein kinase signalling pathways, parasite proliferation was evaluated in the presence of several protein kinase (PK) inhibitors. We found that only KN-93, a classical inhibitor of calcium-calmodulin-dependent kinases (CaMKs), blocked heme-induced cell proliferation. KN-92, an inactive analogue of KN-93, was not able to block this effect. A T. cruzi CaMKII homologue is most likely the main enzyme involved in this process since parasite proliferation was also blocked when Myr-AIP, an inhibitory peptide for mammalian CaMKII, was included in the cell proliferation assay. Moreover, CaMK activity increased in parasite cells with the addition of heme as shown by immunological and biochemical assays. In conclusion, the present results are the first strong indications that CaMKII is involved in the heme-induced cell signalling pathway that mediates parasite proliferation.

  4. Proinflammatory Responses of Heme in Alveolar Macrophages: Repercussion in Lung Hemorrhagic Episodes

    Directory of Open Access Journals (Sweden)

    Rafael L. Simões


    Full Text Available Clinical and experimental observations have supported the notion that free heme released during hemorrhagic and hemolytic episodes may have a major role in lung inflammation. With alveolar macrophages (AM being the main line of defense in lung environments, the influence of free heme on AM activity and function was investigated. We observed that heme in a concentration range found during hemolytic episodes (3–30 μM elicits AM to present a proinflammatory profile, stimulating reactive oxygen species (ROS and nitric oxide (NO generation and inducing IL-1β, IL-6, and IL-10 secretion. ROS production is NADPH oxidase-dependent, being inhibited by DPI and apocynin, and involves p47 subunit phosphorylation. Furthermore, heme induces NF-κB nuclear translocation, iNOS, and also HO-1 expression. Moreover, AM stimulated with free heme show enhanced phagocytic and bactericidal activities. Taken together, the data support a dual role for heme in the inflammatory response associated with lung hemorrhage, acting as a proinflammatory molecule that can either act as both an adjuvant of the innate immunity and as an amplifier of the inflammatory response, leading tissue injury. The understanding of heme effects on pulmonary inflammatory processes can lead to the development of new strategies to ameliorate tissue damage associated with hemorrhagic episodes.

  5. XAS study of the active site of a bacterial heme-sensor

    Energy Technology Data Exchange (ETDEWEB)

    Della Longa, S [Dipartimento di Medicina Sperimentale, Universita dell' Aquila via Vetoio, loc. Coppito II 67100 L' Aquila (Italy); Arcovito, A [Istituto di Biochimica e Biochimica Clinica, Universita Cattolica del Sacro Cuore, Largo F. Vito 1, 00168, Roma (Italy); Brunori, M; Castiglione, N; Cutruzzola, F; Giardina, G; Rinaldo, S [Dipartimento di Scienze Biochimiche ' A. Rossi Fanelli' , Sapienza Universit/a di Roma, P. le A. Moro 5, 00185 Roma (Italy); D' Angelo, P, E-mail: dlonga@caspur.i [Dipartimento di Chimica, Sapienza Universita di Roma, P. le A.Moro 5, 00185 Roma (Italy)


    Denitrifying bacteria control NO and NO{sub 2} cytosolic levels by regulating the expression of denitrification gene clusters via REDOX signalling of specific transcriptional factors that may act as NO sensors in vivo. A protein belonging to the subclass DNR (dissimilative nitrate respiration regulator) from Pseudomonas aeruginosa has been recently suggested to be a heme containing protein. Very recently the three dimensional structure of the apo-form of DNR (in the absence of heme) has been determined by X-Ray crystallography, whereas the holo-form (in the presence of heme) has not yet been crystallized. We have investigated the heme local structure in solution of ferric and ferrous holo-DNR by XAS. The Fe K-edge XANES spectrum of the ferric adduct displays typical features of a low-spin hexacoordinate Fe-heme complex, having two histidines ligated. After chemical reduction, relevant changes of the XANES fingerprints suggest a repositioning of the heme inside the hydrophobic core of the protein in agreement with previously reported structural and spectroscopic evidence. Partial release of the axial ligands leaves the Fe(II)heme available, and very reactive, to bind exogenous ligands like NO, thus supporting its role as the cofactor involved in NO sensing activity.

  6. Mutational analysis of hemoglobin binding and heme utilization by a bacterial hemoglobin receptor. (United States)

    Fusco, W G; Choudhary, N R; Council, S E; Collins, E J; Leduc, I


    Iron is an essential nutrient for most living organisms. To acquire iron from their environment, Gram-negative bacteria use TonB-dependent transporters that bind host proteins at the bacterial surface and transport iron or heme to the periplasm via the Ton machinery. TonB-dependent transporters are barrel-shaped outer membrane proteins with 22 transmembrane domains, 11 surface-exposed loops, and a plug domain that occludes the pore. To identify key residues of TonB-dependent transporters involved in hemoglobin binding and heme transport and thereby locate putative protective epitopes, the hemoglobin receptor of Haemophilus ducreyi HgbA was used as a model of iron/heme acquisition from hemoglobin. Although all extracellular loops of HgbA are required by H. ducreyi to use hemoglobin as a source of iron/heme, we previously demonstrated that hemoglobin binding by HgbA only involves loops 5 and 7. Using deletion, substitution, and site-directed mutagenesis, we were able to differentiate hemoglobin binding and heme acquisition by HgbA. Deletion or substitution of the GYEAYNRQWWA region of loop 5 and alanine replacement of selected histidines affected hemoglobin binding by HgbA. Conversely, mutation of the phenylalanine in the loop 7 FRAP domain or substitution of the NRQWWA motif of loop 5 significantly abrogated utilization of heme from hemoglobin. Our findings show that hemoglobin binding and heme utilization by a bacterial hemoglobin receptor involve specific motifs of HgbA.

  7. Distinct mechanisms for DNA cleavage by myoglobin with a designed heme active center. (United States)

    Zhao, Yuan; Du, Ke-Jie; Gao, Shu-Qin; He, Bo; Wen, Ge-Bo; Tan, Xiangshi; Lin, Ying-Wu


    Heme proteins perform diverse biological functions, of which myoglobin (Mb) is a representative protein. In this study, the O2 carrier Mb was shown to cleave double stranded DNA upon aerobic dithiothreitol-induced reduction, which is fine-tuned by an additional distal histidine, His29 or His43, engineered in the heme active center. Spectroscopic (UV-vis and EPR) and inhibition studies suggested that free radicals including singlet oxygen and hydroxyl radical are responsible for efficient DNA cleavage via an oxidative cleavage mechanism. On the other hand, L29E Mb, with a distinct heme active center involving three water molecules in the met form, was found to exhibit an excellent DNA cleavage activity that was not depending on O2. Inhibition and ligation studies demonstrated for the first time that L29E Mb cleaves double stranded DNA into both the nicked circular and linear forms via a hydrolytic cleavage mechanism, which resembles native endonucleases. This study provides valuable insights into the distinct mechanisms for DNA cleavage by heme proteins, and lays down a base for creating artificial DNA endonucleases by rational design of heme proteins. Moreover, this study suggests that the diverse functions of heme proteins can be fine-tuned by rational design of the heme active center with a hydrogen-bonding network.

  8. Synthesis and self-assembly of Chitosan-g-Polystyrene copolymer: A new route for the preparation of heavy metal nanoparticles

    KAUST Repository

    Francis, Raju S.


    Amphiphilic graft copolymers made of a Chitosan (CS) backbone and three arm polystyrene (PS) grafts were prepared by "grafting onto" strategy using Toluene Diisocyanate. IR spectroscopy and SEC show the successful grafting process. SEM pictures of Chitosan-g-Polystyrene (CS-g-PS) indicate a spherulite like surface and exhibit properties that result from the disappearance of Chitosan crystallinity. The introduced polystyrene star grafts units improve hydrophobic properties considerably as confirmed by the very high solubility of (CS-g-PS) in organic solvents. The graft copolymer which self-assembles into polymeric micelles in organic media demonstrates much better adsorption of transition and inner transition metal ions than pure Chitosan whose amine groups are not necessarily available due to crystallinity.

  9. Synthesis and self-assembly of chitosan-g-polystyrene copolymer: a new route for the preparation of heavy metal nanoparticles. (United States)

    Francis, Raju; Baby, Deepa K; Gnanou, Yves


    Amphiphilic graft copolymers made of a Chitosan (CS) backbone and three arm polystyrene (PS) grafts were prepared by "grafting onto" strategy using Toluene Diisocyanate. IR spectroscopy and SEC show the successful grafting process. SEM pictures of Chitosan-g-Polystyrene (CS-g-PS) indicate a spherulite like surface and exhibit properties that result from the disappearance of Chitosan crystallinity. The introduced polystyrene star grafts units improve hydrophobic properties considerably as confirmed by the very high solubility of (CS-g-PS) in organic solvents. The graft copolymer which self-assembles into polymeric micelles in organic media demonstrates much better adsorption of transition and inner transition metal ions than pure Chitosan whose amine groups are not necessarily available due to crystallinity.

  10. Facile synthesis of 3D hierarchical foldaway-lantern-like LiMnPO4 by nanoplate self-assembly, and electrochemical performance for Li-ion batteries. (United States)

    Chen, Dezhi; Wei, Wei; Wang, Ruining; Lang, Xiu-Feng; Tian, Yu; Guo, Lin


    Olivine-structured LiMnPO(4) with 3D foldaway-lantern-like hierarchical structures have been prepared via a one-step, template-free, solvothermal approach in ethylene glycol. The foldaway-lantern-like LiMnPO(4) microstructures are composed of numerous nanoplates with thickness of about 20 nm. A series of electron microscopy characterization results indicate that the obtained primary LiMnPO(4) nanoplates are single crystalline in nature, growing along the [010] direction in the (100) plane. Time-dependent morphology evolution suggests that ethylene glycol plays dual roles in oriented growth and self-assembly of such unique structures. After carbon coating, the as-prepared LiMnPO(4) cathode demonstrated a flat potential at 4.1 V versus Li/Li(+) with a specific capacity close to 130 mA h g(-1) at 0.1 C, along with excellent cycling stability.

  11. SEMICONDUCTOR DEVICES Density-controllable nonvolatile memory devices having metal nanocrystals through chemical synthesis and assembled by spin-coating technique (United States)

    Guangli, Wang; Yubin, Chen; Yi, Shi; Lin, Pu; Lijia, Pan; Rong, Zhang; Youdou, Zheng


    A novel two-step method is employed, for the first time, to fabricate nonvolatile memory devices that have metal nanocrystals. First, size-averaged Au nanocrystals are synthesized chemically; second, they are assembled into memory devices by a spin-coating technique at room temperature. This attractive approach makes it possible to tailor the diameter and control the density of nanocrystals individually. In addition, processes at room temperature prevent Au diffusion, which is a main concern for the application of metal nanocrystal-based memory. The experimental results, both the morphology characterization and the electrical measurements, reveal that there is an optimum density of nanocrystal monolayer to balance between long data retention and a large hysteresis memory window. At the same time, density-controllable devices could also feed the preferential emphasis on either memory window or retention time. All these facts confirm the advantages and novelty of our two-step method.

  12. Microwave-assisted synthesis of self-assembled BiO{sub 1.84}H{sub 0.08} hierarchical nanostructures as a new photocatalyst

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Pei; Hou, Dongfang; Shi, Hongyu; Chen, Chaoji; Huang, Yunhui; Hu, Xianluo, E-mail:


    Highlights: • Hierarchical BiO{sub 1.84}H{sub 0.08} nanostructures were prepared by microwave irradiation. • The microwave-hydrothermal method is rapid, one-pot, and low-temperature. • The morphology of the BiO{sub 1.84}H{sub 0.08} nanostructures can be tailored. • The photocatalytic properties were studied upon UV–visible irradiation. • Self-assembled flower-like BiO{sub 1.84}H{sub 0.08} shows enhanced photocatalytic activity. - Abstract: Self-assembled hierarchical nanostructures of BiO{sub 1.84}H{sub 0.08} spheres have been successfully synthesized by a rapid, one-pot, low-temperature hydrothermal route under microwave irradiation. The as-prepared hierarchically nanostructured BiO{sub 1.84}H{sub 0.08} possesses a high surface area of 113 m{sup 2} g{sup −1}. By adjusting the reaction temperature and reactant concentration, the morphology of the as-formed BiO{sub 1.84}H{sub 0.08} nanostructures can be tailored. As a new photocatalyst, the flower-like spherical BiO{sub 1.84}H{sub 0.08} exhibits enhanced photocatalytic activity for degradation of Rhodamine B (RhB) under UV–visible light. The synthetic procedure is simple, efficient, and scalable for mass production. Therefore, the as-formed BiO{sub 1.84}H{sub 0.08} hierarchical nanostructures may offer great potential applications for decomposition of organic contaminants.

  13. Biologically Inspired Self-assembling Synthesis of Bone-like Nano-hydroxyapatite/PLGA- (PEG-ASP)n Composite: A New Biomimetic Bone Tissue Engineering Scaffold Material

    Institute of Scientific and Technical Information of China (English)


    A new biomimetic bone tissue engineering scaffold material, nano-HA/ PLGA-( PEG- ASP )n composite, was synthesized by a biologically inspired self assembling approach. A novel biodegradable PLGA( PEG-ASP ) n copolymer with pendant amine functional groups and enhanced hydrophilicity was synthesized by bulk ring-opening copolymerization by DL-lactide( DLLA ) and glycolide( GA ) with Aspartic acid ( ASP )-Polyethylene glycol( PEG ) alt-prepolymer. A Three-dimensional, porous scaffold of the PLGA-( PEG-ASP )n copolymer was fabricated by a solvent casting, particulate leaching process. The scaffold was then incubated in modified simulated body fluid ( mSBF ) . Growth of HA nanocrystals on the inner pore surfaces of the porous scaffold is confirmed by calcium ion binding analyses, SEM, mass increase measurements and quantification of phosphate content within scaffolds . SEM analysis demonstrated the nucleation and growth of a continuous bonelike, low crystalline carbonated HA nanocrystals on the inner pore surfawes of the PLGA-( PEG-ASP)n scaffolds. The amount of calcium binding, total mass and the mass of pbosphate on experimental PLGA-( PEG- ASP )n scaffolds at different incubation times in mSBF was significantly greater than that of control PLGA scaffolds . This nano-HA/ PLGA- ( PEG-ASP )n composite shows some features of natural bone both in main composition and hierarchical microstructure. The ASPPEG alt-prepolymer modified PLGA copolymer provide a controllable high surface density and distribution of anionic functional groups which would enhauce nucleation and growth of bonelike mineral following exposure to mSBF. This biomimetic treatment provides a simple method for surface funetionalization and subsequent mineral nucleation and self-assembling on biodegradable polymer scaffolds for tissue engineering.

  14. Synthesis and self-assembly of amphiphilic poly(acrylicacid)-poly(ɛ-caprolactone)-poly(acrylicacid) block copolymer as novel carrier for 7-ethyl-10-hydroxy camptothecin. (United States)

    Djurdjic, Beti; Dimchevska, Simona; Geskovski, Nikola; Petrusevska, Marija; Gancheva, Valerya; Georgiev, Georgi; Petrov, Petar; Goracinova, Katerina


    The process of molecular self-assembly plays a crucial role in formulation of polymeric nanoparticulated drug delivery carriers as it creates the possibility for enhanced drug encapsulation and carrier surface engineering. This study aimed to develop a novel self-assembled polymeric micelles for targeted delivery in tumor cells in order to overcome not only various drawbacks of 7-ethyl-10-hydroxy camptothecin (SN-38) but also various reported limitations of other drug delivery systems, especially low drug loading and premature release. Custom synthesized amphiphilic triblock copolymer poly(acrylic acid)-poly(ɛ-caprolactone)-poly(acrylic acid) (PAA(13)-PCL(35)-PAA(13)) was used to prepare kinetically stable micelles by nanoprecipitation and modified nanoprecipitation procedure. Core-shell micelles with diameter of 120-140 nm, negative zeta potential and satisfactory drug loading were produced. The prepared formulations were stable in pH range of 3-12 and in media with NaCl concentration <1 mol/l. Screening mixed level factorial 3 × 2(2) design identified that the process temperature as well as the type of organic solvent has influence upon the efficacy of encapsulation, particle size, dissolution rate and burst release. Fourier transform infrared and differential scanning calorimetry analyses confirmed the entrapment of the active substance into the micelles. The kinetic analysis of dissolution studies revealed that the main mechanism of drug release from the prepared formulations is Fickian diffusion. Growth inhibition studies as well as DNA fragmentation assay performed on SW-480 cell lines clearly demonstrated increased growth inhibition effect and presence of fragmented DNA in cells treated with loaded micelles compared to SN-38 solution. Altogether, these results point out to potential biomedical and clinical application of PAA-PCL-PAA systems in the future.

  15. Metal-organic and supramolecular networks driven by 5-chloronicotinic acid: Hydrothermal self-assembly synthesis, structural diversity, luminescent and magnetic properties

    Energy Technology Data Exchange (ETDEWEB)

    Gao, Zhu-Qing, E-mail: [School of Chemical and Biological Engineering, Taiyuan University of Science and Technology, Taiyuan 030021 (China); Li, Hong-Jin [School of Chemical and Biological Engineering, Taiyuan University of Science and Technology, Taiyuan 030021 (China); Gu, Jin-Zhong, E-mail: [College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000 (China); Zhang, Qing-Hua [School of Chemical and Biological Engineering, Taiyuan University of Science and Technology, Taiyuan 030021 (China); Kirillov, Alexander M. [Centro de Química Estrutural, Complexo I, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049–001 Lisbon (Portugal)


    Four new crystalline solids, namely [Co{sub 2}(µ{sub 2}-5-Clnic){sub 2}(µ{sub 3}-5-Clnic){sub 2}(µ{sub 2}-H{sub 2}O)]{sub n} (1), [Co(5-Clnic){sub 2}(H{sub 2}O){sub 4}]·2(5-ClnicH) (2), [Pb(µ{sub 2}-5-Clnic){sub 2}(phen)]{sub n} (3), and [Cd(5-Clnic){sub 2}(phen){sub 2}]·3H{sub 2}O (4) were generated by hydrothermal self-assembly methods from the corresponding metal(II) chlorides, 5-chloronicotinic acid (5-ClnicH) as a principal building block, and 1,10-phenanthroline (phen) as an ancillary ligand (optional). All the products 1–4 were characterized by IR spectroscopy, elemental analysis, thermogravimetric (TGA), powder X-ray diffraction (PXRD) and single-crystal X-ray diffraction. Their structures range from an intricate 3D metal-organic network 1 with the 3,6T7 topology to a ladder-like 1D coordination polymer 3 with the 2C1 topology, whereas compounds 2 and 4 are the discrete 0D monomers. The structures of 2 and 4 are further extended (0D→2D or 0D→3D) by hydrogen bonds, generating supramolecular networks with the 3,8L18 and ins topologies, respectively. Synthetic aspects, structural features, thermal stability, magnetic (for 1) and luminescent (for 3 and 4) properties were also investigated and discussed. - Graphical abstract: A new series of crystalline solids was self-assembled and fully characterized; their structural, topological, luminescent and magnetic features were investigated. Display Omitted.

  16. Heme as a danger molecule in pathogen recognition. (United States)

    Wegiel, Barbara; Hauser, Carl J; Otterbein, Leo E


    Appropriate control of redox mechanisms are critical for and effective innate immune response, which employs multiple cell types, receptors and molecules that recognize danger signals when they reach the host. Recognition of pathogen-associated pattern molecules (PAMPs) is a fundamental host survival mechanism for efficient elimination of invading pathogens and resolution of the infection and inflammation. In addition to PAMPs, eukaryotic cells contain a plethora of intracellular molecules that are normally secured within the confines of the plasma membrane, but if liberated and encountered in the extracellular milieu can provoke rapid cell activation. These are known as Alarmins or Danger-Associated Molecular Patterns (DAMPs) and can be released actively by cells or passively as a result of sterile cellular injury after trauma, ischemia, or toxin-induced cell rupture. Both PAMPs and DAMPs are recognized by a series of cognate receptors that increase the generation of free radicals and activate specific signaling pathways that result in regulation of a variety of stress response, redox sensitive genes. Multiple mediators released, as cells die include, but are not limited to ATP, hydrogen peroxide, heme, formyl peptides, DNA or mitochondria provide the second signal to amplify immune responses. In this review, we will focus on how sterile and infective stimuli activate the stress response gene heme oxygenase-1 (Hmox1, HO-1), a master gene critical to an appropriate host response that is now recognized as one with enormous therapeutic potential. HO-1 gene expression is regulated in large part by redox-sensitive proteins including but not limited to nrf2. Both PAMPs and DAMPs increase the activation of nrf2 and HO-1. Heme is a powerful pro-oxidant and as such should be qualified as a DAMP. With its degradation by HO-1a molecule of carbon monoxide (CO) is generated that in turn serves as a bioactive signaling molecule. PAMPs such as bacterial endotoxin activate HO-1

  17. Assembly of 4-, 6- and 8-connected Cd(II) pseudo-polymorphic coordination polymers: Synthesis, solvent-dependent structural variation and properties (United States)

    Li, Zhao-Hao; Xue, Li-Ping; Miao, Shao-Bin; Zhao, Bang-Tun


    The reaction of Cd(NO3)2·4H2O, 2,5-thiophenedicarboxylic acid (H2tdc) and 1,2-bis(imidazol-1‧-yl)methane (bimm) by modulating solvent systems yielded three highly connected pseudo-polymorphic coordination polymers based on different dinuclear [Cd2(CO2)2] subunits bridged by carboxylate groups. Single crystal structural analyses reveal structural variation from 4-connected 2D sql layer, 6-connected 2-fold interpenetrated 3D pcu to 8-connected 3D bcu-type network in compounds 1-3. The structural dissimilarity in the structures dependent on the coordination environments of Cd(II) ions and linking modes of mixed ligand influenced by different solvent systems during the synthesis process. Moreover, thermogravimetric and photoluminescence behaviors of 1-3 were also investigated for the first time, and all the complexes emit blue luminescence in the solid state.

  18. Self-assembled monolayers of Aβ peptides on Au electrodes: an artificial platform for probing the reactivity of redox active metals and cofactors relevant to Alzheimer's disease. (United States)

    Pramanik, Debajyoti; Sengupta, Kushal; Mukherjee, Soumya; Dey, Somdatta Ghosh; Dey, Abhishek


    The water-soluble hydrophilic part of human Aβ peptide has been extended to include a C-terminal cysteine residue. Utilizing the thiol functionality of this cysteine residue, self-assembled monolayers (SAM) of these peptides are formed on Au electrodes. Atomic force microscopy imaging confirms formation of small Aβ aggregates on the surface of the electrode. These aggregates bind redox active metals like Cu and cofactors like heme, both of which are proposed to generate toxic partially reduced oxygen species (PROS) and play a vital role in Alzheimer's disease. The spectroscopic and electrochemical properties of these Cu and heme bound Aβ SAM are similar to those reported for the soluble Cu and heme bound Aβ peptide. Experiments performed on these Aβ-SAM electrodes clearly demonstrate that (1) heme bound Aβ is kinetically more competent in reducing O(2) than Cu bound Aβ, (2) under physiological conditions the reduced Cu site produces twice as much PROS (measured in situ) than the reduced heme site, and (3) chelators like clioquinol remove Cu from these aggregates, while drugs like methylene blue inhibit O(2) reactivity of the heme cofactor. This artificial construct provides a very easy platform for investigating potential drugs affecting aggregation of human Aβ peptides and PROS generation by its complexes with redox active metals and cofactors.

  19. Chemical synthesis and assembly of quasi-two-dimensional metal chalcogenides graphene analogues%准二维金属硫属化合物类石墨烯结构的化学合成与组装

    Institute of Scientific and Technical Information of China (English)

    冯冯; 冯骏; 吴长征; 谢毅


    Inorganic graphene analogues are regarded as a typical quasi-two-dimensional nanostructure system. Graphene analogues entail more structural parameters that could be controlled compared with the pure-carbon graphene, which leads to the well controlled energy-band type and gap width, showing the promising signs in constructing electronic device in the field of energy conversion and energy storage. In this regard, metal chalcogenides (MCs) have drawn extensive attention as an important graphene analogue material system due to its unique quasi-two dimensional crystallographic structure. Herein, we reviewed the synthesis and assembly methodologies for graphene analogues of MCs, as well as their functional applications of their assembled nanostructures. We put forward the strategies that utilizing chemical methods to weaken interlayer force and embodying the highly anisotropic characteristics in the internal structural lattices to successfully realize the fabrication of graphene analogues. Finally, we also summarized the assembly strategy of graphene analogues and their promising applications in the area of energy storage and intelligent responsiveness nanodevices.%无机类石墨烯属于准二维纳米结构体系,因其具有比纯碳石墨烯本身更多的调控参数,比如带隙类型及其带隙宽度可调节等,在电子器件构筑和能量转化存储等领域有着重要的科学意义和广阔的应用前景.近年来,具有准二维特征的金属硫属化合物(metal chalcogenides,MCs)作为类石墨烯结构的重要材料体系受到了广泛关注.本文概述了近年来金属硫属化合物类石墨烯结构的化学制备方法及其可能的组装应用,提出了通过系列方法影响层间作用力及利用晶体各向异性等材料设计与合成策略来实现类石墨烯的化学合成,并展望了类石墨烯的组装结构在能量存储与智能传感领域的应用前景.

  20. A simple method for the determination of reduction potentials in heme proteins. (United States)

    Efimov, Igor; Parkin, Gary; Millett, Elizabeth S; Glenday, Jennifer; Chan, Cheuk K; Weedon, Holly; Randhawa, Harpreet; Basran, Jaswir; Raven, Emma L


    We describe a simple method for the determination of heme protein reduction potentials. We use the method to determine the reduction potentials for the PAS-A domains of the regulatory heme proteins human NPAS2 (Em=-115 mV ± 2 mV, pH 7.0) and human CLOCK (Em=-111 mV ± 2 mV, pH 7.0). We suggest that the method can be easily and routinely applied to the determination of reduction potentials across the family of heme proteins.

  1. Heme oxygenase 1 transgenic mice as a model to study neuroprotection. (United States)

    Maines, Mahin D


    Bile pigments and CO are formed in the course of heme degradation by the isozymes and are biologically active moieties. In the course of heme degradation the chelated iron is also released. Heme and iron are prooxidants, whereas bile pigments are antioxidants. In addition, CO functions as a signal molecule and HO-2 may serve as an intracellular "sink" for NO. In the balance, the published data suggest that the HO system functions in cellular defense mechanisms. The methods described in this chapter can be used to assess the tissue/cell toxicity of chemicals in general, and as pertains to the defense activity of HO-1, specifically.

  2. Self assembly of interlocked architectures

    CERN Document Server

    Schergna, S


    An area of great interest is the synthesis and characterisation of molecules possessing moving parts, with the goal that they can act as 'molecular machine' carrying out tasks that molecules with fixed conventional architectures cannot do. Rotaxanes and catenanes (mechanically interlocked architectures) represent one approach toward achieving these aims as their component wheels and / or threads are connected together but can still move, in certain, controlled directions. This thesis focused on the study of structural rigidity and the preorganisation of thread binding sites as factors of major influence on template efficiency in the synthesis of hydrogen bond assembled supramolecular structures (rotaxanes and catenanes). Chapter One gives a brief outline of the common synthetic approaches to interlocked architectures (catenanes and rotaxanes) that are now being developed to address the problems outlined above. Chapter Two and Chapter Three concerns the synthesis of novel amide-based rotaxanes containing vario...

  3. Heme Oxygenase-1 Promotes Delayed Wound Healing in Diabetic Rats. (United States)

    Chen, Qing-Ying; Wang, Guo-Guang; Li, Wei; Jiang, Yu-Xin; Lu, Xiao-Hua; Zhou, Ping-Ping


    Diabetic ulcers are one of the most serious and costly chronic complications for diabetic patients. Hyperglycemia-induced oxidative stress may play an important role in diabetes and its complications. The aim of the study was to explore the effect of heme oxygenase-1 on wound closure in diabetic rats. Diabetic wound model was prepared by making an incision with full thickness in STZ-induced diabetic rats. Wounds from diabetic rats were treated with 10% hemin ointment for 21 days. Increase of HO-1 protein expression enhanced anti-inflammation and antioxidant in diabetic rats. Furthermore, HO-1 increased the levels of VEGF and ICAM-1 and expressions of CBS and CSE protein. In summary, HO-1 promoted the wound closure by augmenting anti-inflammation, antioxidant, and angiogenesis in diabetic rats.

  4. Heme Oxygenase-1 Promotes Delayed Wound Healing in Diabetic Rats

    Directory of Open Access Journals (Sweden)

    Qing-Ying Chen


    Full Text Available Diabetic ulcers are one of the most serious and costly chronic complications for diabetic patients. Hyperglycemia-induced oxidative stress may play an important role in diabetes and its complications. The aim of the study was to explore the effect of heme oxygenase-1 on wound closure in diabetic rats. Diabetic wound model was prepared by making an incision with full thickness in STZ-induced diabetic rats. Wounds from diabetic rats were treated with 10% hemin ointment for 21 days. Increase of HO-1 protein expression enhanced anti-inflammation and antioxidant in diabetic rats. Furthermore, HO-1 increased the levels of VEGF and ICAM-1 and expressions of CBS and CSE protein. In summary, HO-1 promoted the wound closure by augmenting anti-inflammation, antioxidant, and angiogenesis in diabetic rats.

  5. Design and synthesis of collagen mimetic peptide derivatives for studying triple helix assembly and collagen mimetic peptide-collagen binding interaction (United States)

    Mo, Xiao


    Collagen is the principal tensile clement of the extra-cellular matrix in mammals and is the basic scaffold for cells and tissues. Collagen molecules are comprised of homo-trimeric helices (e.g. collagen type II and type III), ABB type hetero-trimeric helices (e.g. collagen type I, type IV, and type V), or ABC type hetero-trimeric helices (e.g. type V). Mimicry of collagen structures can help elucidate collagen triple helical conformation and provide insights into making novel collagen-like biomaterials. Our group previously reported a new physical collagen modification method, which was based on non-covalent interaction between collagen mimetic peptide (CMP: -(Pro-Hyp-Gly) x-) and natural collagen. We hypothesized that CMP binds to collagen through a process involving both strand invasion and triple helix assembly. The aim of this dissertation is to study structural formation and stability of collagen triple helix, and to investigate CMP-collagen binding interactions using two types of CMP derivatives: covalently templated CMP trimer and CMP-nanoparticle conjugates. We demonstrated that covalently templated ABB type CMP hetero-trimers could be prepared by a versatile synthetic strategy involving both solid phase and solution peptide coupling. Our thermal melting studies showed that the templated CMP hetero-trimers formed collagen-like triple helices and their folding kinetics correlated with the amino acid compositions of the individual CMP strands. We also studied the thermal melting behavior and folding kinetics of a templated hetero-trimer complex comprised of CMP and a peptide derived from collagen. This synthetic strategy can be readily extended to synthesize other ABB type hetero-trimers to investigate their local melting behavior and biological activity. We also prepared colloidally stable CMP functionalized gold nanoparticles (Au-CMPs) as a TEM marker for investigating the CMP-collagen interaction. Au-CMP showed preferential binding to collagen fiber's gap

  6. Role of heme oxygenase 1 in TNF/TNF receptor-mediated apoptosis after hepatic ischemia/reperfusion in rats. (United States)

    Kim, Seok-Joo; Eum, Hyun-Ae; Billiar, Timothy R; Lee, Sun-Mee


    Hepatocellular apoptosis commonly occurs in ischemia/reperfusion (I/R) injury. The binding of tumor necrosis factor (TNF) to TNF receptor 1 (TNFR1) leads to the formation of a death-inducing signaling complex (DISC), which subsequently initiates a caspase cascade resulting in apoptosis. Heme oxygenase 1 (HO-1) confers cytoprotection against cell death in I/R injury and inhibits stress-induced apoptotic pathways in vitro. This study investigated the role of HO-1 in modulating TNF/TNFR1-mediated cell death pathways in hepatic I/R injury. Rats were pretreated with hemin, an HO-1 inducer, and zinc protoporphyrin (ZnPP), an HO-1 inhibitor, before undergoing hepatic I/R. Heme oxygenase 1 activity increased after reperfusion. Ischemia/reperfusion-induced hepatocellular apoptosis was attenuated by hemin, as determined by the caspase-3 and -8 activity assays and TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling). Zinc protoporphyrin eliminated the cytoprotective effect of hemin. Hepatic TNFR1 protein expression was unchanged among the experimental groups, whereas mitochondrial TNFR1 protein increased after I/R. Ischemia/reperfusion increased the quantity of DISC components, including TRADD (TNFR1-associated death domain), FADD (Fas-associated death domain), and caspase-8, as well as the assembly of DISCs within the liver. In the mitochondrial fraction, TNFR1-associated caspase-8 was increased after I/R. These increases were attenuated by hemin; zinc protoporphyrin eliminated this effect. Our findings suggest that the cytoprotective effects of HO-1 are mediated by suppression of TNF/TNFR1-mediated apoptotic signaling, specifically by modulating apoptotic DISC formation and mitochondrial TNFR1 translocation during hepatic I/R.

  7. Assembling consumption

    DEFF Research Database (Denmark)

    Assembling Consumption marks a definitive step in the institutionalisation of qualitative business research. By gathering leading scholars and educators who study markets, marketing and consumption through the lenses of philosophy, sociology and anthropology, this book clarifies and applies...... the investigative tools offered by assemblage theory, actor-network theory and non-representational theory. Clear theoretical explanation and methodological innovation, alongside empirical applications of these emerging frameworks will offer readers new and refreshing perspectives on consumer culture and market...... societies. This is an essential reading for both seasoned scholars and advanced students of markets, economies and social forms of consumption....

  8. Scalable synthesis of hierarchical hollow Li4Ti5O12 microspheres assembled by zigzag-like nanosheets for high rate lithium-ion batteries (United States)

    Zhu, Kunxu; Gao, Hanyang; Hu, Guoxin; Liu, Mengjing; Wang, Haochen


    Electrochemical performance, abundance and cost are three crucial criteria to comprehensively evaluate the feasibility of Li4Ti5O12 as an electrode material for lithium-ion batteries (LIBs). Herein, hierarchical hollow Li4Ti5O12 microspheres (HLTOMs) assembled by zigzag-like nanosheets are synthesized by hydrothermal treatment of scalable lithium peroxotitanate complex solution using low-cost commercial H2TiO3 particles as titanium sources, followed by a calcination treatment. Precursor solution concentration, Li/Ti ratio, hydrothermal temperature and duration are found correlative and should be optimized to obtain pure Li4Ti5O12 products. A high yield of HLTOMs up to 120 g L-1 was achieved. Due to the unique morphology, the HLTOMs deliver an outstanding rate capability of 139, 125 and 108 mA h g-1 at 10, 20 and 30 C, respectively, and exhibit 94% capacity retention after 1000 cycles at 30C indicating excellent stability. These values are much superior to those of commercial Li4Ti5O12 particles (CLTOPs), showing HLTOMs are promising anode materials for LIBs.

  9. Synthesis, properties and surface self-assembly of a pentanuclear cluster based on the new π-conjugated TTF-triazole ligand (United States)

    Cui, Long; Geng, Yan-Fang; Leong, Chanel F.; Ma, Qian; D'Alessandro, Deanna M.; Deng, Ke; Zeng, Qing-Dao; Zuo, Jing-Lin


    The new π-extended redox-active ligand with both TTF and triazole units, 6-(4,5-bis(propylthio)-1,3-dithiol-2-ylidene)-1H-[1,3]dithiolo[4‧,5‧:4,5]benzo [1,2-d] [1-3]triazole, has been successfully prepared. Based on the versatile ligand and Cu(tta)2 precursors (tta- = 4,4,4-trifluoro-1-(thiophen-2-yl)butane-1,3-dione), a TTF-based pentanuclear CuII cluster (Cu5(tta)4(TTFN3)6) is synthesized and structurally characterized. Their absorption and electrochemical properties are investigated. Antiferromagnetic couplings are operative between metal ion centers bridged by triazoles in the complex. The self-assembled structure of the cluster complex on a highly oriented pyrolytic graphite (HOPG) surface was observed using scanning tunneling microscopy and density functional theory (DFT) calculations have been performed to provide insight into the formation mechanism. The introduction of the redox-active TTF unit into the cluster complexes with interesting magnetic properties renders them promising candidates for new multifunctional materials.

  10. Synthesis, molecular docking and biological evaluation of 1-phenylsulphonyl-2-(1-methylindol-3-yl)-benzimidazole derivatives as novel potential tubulin assembling inhibitors. (United States)

    Wang, Yan-Ting; Cai, Xun-Chao; Shi, Tian-Qi; Zhang, Ya-Liang; Wang, Zhong-Chang; Liu, Chang-Hong; Zhu, Hai-Liang


    A series of new 1-phenylsulphonyl-2-(1-methylindol-3-yl)-benzimidazole derivatives were designed, synthesized and evaluated as potential inhibitors of tubulin polymerization and anthropic cancer cell lines. Among them, compound 33 displayed the most potent tubulin polymerization inhibitory activity in vitro (IC50  = 1.41 μM) and strong antiproliferative activities against A549, Hela, HepG2 and MCF-7 cell lines in vitro with GI50 value of 1.6, 2.7, 2.9 and 4.3 μM, respectively, comparable with the positive control colchicine (GI50 value of 4.1, 7.2, 9.5 and 14.5 μM, respectively) and CA-4 (GI50 value of 2.2, 4.3, 6.4 and 11.4 μM, respectively). Simultaneously, we evaluated that compound 33 could effectively induce apoptosis of A549 associated with G2/M phase cell cycle arrest. Immunofluorescence microscopy also clearly indicated compound 33 a potent antimicrotubule agent. Docking simulation showed that compound 33 could bind tightly with the colchicine-binding site and act as a tubulin inhibitor. Three-dimensional-QSAR model was also built to provide more pharmacophore understanding that could be used to design new agents with more potent tubulin assembling inhibitory activity in the future.

  11. Metal-organic and supramolecular networks driven by 5-chloronicotinic acid: Hydrothermal self-assembly synthesis, structural diversity, luminescent and magnetic properties (United States)

    Gao, Zhu-Qing; Li, Hong-Jin; Gu, Jin-Zhong; Zhang, Qing-Hua; Kirillov, Alexander M.


    Four new crystalline solids, namely [Co2(μ2-5-Clnic)2(μ3-5-Clnic)2(μ2-H2O)]n (1), [Co(5-Clnic)2(H2O)4]·2(5-ClnicH) (2), [Pb(μ2-5-Clnic)2(phen)]n (3), and [Cd(5-C