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Sample records for bisphosphate carboxylase small

  1. Soybean ribulose bisphosphate carboxylase small subunit: Mechanisms and determinants of RNA turnover. Annual progress report

    Energy Technology Data Exchange (ETDEWEB)

    Meagher, R.B. [Georgia Univ., Athens, GA (United States). Dept. of Genetics

    1993-12-31

    An in vitro degradation system has been developed from petunia and soybean polysomes in order to investigate the mechanisms and determinants controlling RNA turnover in higher plants. This system faithfully degrades soybean ribulose-1,5-bisphosphate carboxylase small subunit (rbcS) mRNA into the same products observed in total RNA preparations. In previous years it was shown that the most stable products represent a nested constellation of fragments, which are shortened from their 3{prime} ends, and have intact 5{prime} ends. Exogenous rbcS RNA tagged with novel 5{prime} sequence 15 or 56 bp long were synthesized in vitro as Sp6 and T7 runoff transcripts, respectively. When added to the system they were degraded faithfully into constellation of products which were 15 or 56 bp longer than the endogenous products, respectively. Detailed kinetics on the appearance of these exogenous products confirmed degradation proceeds in an overall 3{prime} to 5{prime} direction but suggested that there are multiple pathways through which the RNA may be degraded. To further demonstrate a precursor product relationships, in vitro synthesized transcripts truncated at their 3{prime} ends were shown to degrade into the expected smaller fragments previously mapped in the 5{prime} portion of the rbcS RNA.

  2. A Novel Approach to Functional Analysis of the Ribulose Bisphosphate Carboxylase Small Subunit Gene by Agrobacterium-Mediated Gene Silencing

    Institute of Scientific and Technical Information of China (English)

    Xiao-Fu Zhou; Peng-Da Ma; Ren-Hou Wang; Bo Liu; Xing-Zhi Wang

    2006-01-01

    A novel approach to virus-induced post-transcriptional gene silencing for studying the function of the ribulose bisphosphate carboxylase small subunlt (rbcS) gene was established and optimized using potato virus X vector and Nicotiana benthamiana as experimental material. The analysis of silencing phenomena,transcriptional level, protein expression, and pigment measurement showed that the expression of the rbcS endogenous gene was inactivated by the expression of a 500-bp homologous cDNA fragment carried in the virus vector.

  3. Differential transcription and message stability of two genes encoding soybean ribulose 1,5-bisphosphate carboxylase small subunit

    International Nuclear Information System (INIS)

    The expression of two closely related soybean ribulose bisphosphate carboxylase small subunit (Rubisco ss) genes, SRS1 and SRS4, has been compared. These genes account for approximately 2-4% of the total transcription in light grown leaves, SRS4 being twice as transcriptionally active as SRS1. The transcription of these genes is reduced more than 30 fold after a pulse of far-red light or extended periods of darkness. When etiolated seedlings are shifted to the light the transcription of both genes increases 30-50 fold. Despite this 30-fold range in transcriptional expression the steady state mRNA levels in light and dark grown tissue differ by less than 8 fold. This suggests that the mRNAs are less stable in light grown tissue. 38 refs., 5 figs

  4. Covalent dimerization of ribulose bisphosphate carboxylase subunits by UV radiation

    International Nuclear Information System (INIS)

    The effect of UV radiation (UV-A, UV-B and UV-C) on ribulose bisphosphate carboxylase from a variety of plant species was examined. The exposition of plant leaves or the pure enzyme to UV radiation produced a UV-dependent accumulation of a 65 kDa polypeptide (P65). Different approaches were utilized to elucidate the origin and structure of P65: electrophoretic and fluorographic analyses of 35S-labelled ribulose biphosphate carboxylase exposed to UV radiation and immunological experiments using antibodies specific for P65, for the large and small subunits of ribulose biphosphate carboxylase and for high-molecular-mass aggregates of the enzyme. These studies revealed that P65 is a dimer, formed by the covalent, non-disulphide linkage of one small subunit with one large subunit of ribulose biphosphate carboxylase. For short periods of time (<1 h), the amount of P65 formed increased with the duration of the exposure to the UV radiation and with the energy of the radiation applied. Prolonged exposure to UV radiation (1-6 h) resulted in the formation of high-molecular-mass aggregates of ribulose biphosphate carboxylase. Formation of P65 was shown to depend on the native state of the protein, was stimulated by inhibitors of enzyme activity, and was inhibited by activators of enzyme activity. A UV-independent accumulation of P65 was also achieved by the in vitro incubation of plant crude extracts. However, the UV-dependent and the UV-independent formation of P65 seemed to occur by distinct molecular mechanisms. The UV-dependent accumulation of P65 was immunologically detected in all species examined, including Lemna minor, Arum italicum, Brassica oleracea, Triticum aestivum, Zea mays, Pisum sativum and Phaseolus vulgaris, suggesting that it may constitute a universal response to UV radiation, common to all photosynthetic tissues. (Author)

  5. Covalent dimerization of ribulose bisphosphate carboxylase subunits by UV radiation

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira, R.M.B. [Universidade Tecnica, Lisbon (Portugal). Inst. Superior de Agronomia]|[Universidade Nova de Lisboa, Oeiras (Portugal). Instituto de Tecnologia Quimica e Biologica; Franco, E.; Teixeira, A.R.N. [Universidade Tecnica, Lisbon (Portugal). Inst. Superior de Agronomia

    1996-08-15

    The effect of UV radiation (UV-A, UV-B and UV-C) on ribulose bisphosphate carboxylase from a variety of plant species was examined. The exposition of plant leaves or the pure enzyme to UV radiation produced a UV-dependent accumulation of a 65 kDa polypeptide (P65). Different approaches were utilized to elucidate the origin and structure of P65: electrophoretic and fluorographic analyses of {sup 35}S-labelled ribulose biphosphate carboxylase exposed to UV radiation and immunological experiments using antibodies specific for P65, for the large and small subunits of ribulose biphosphate carboxylase and for high-molecular-mass aggregates of the enzyme. These studies revealed that P65 is a dimer, formed by the covalent, non-disulphide linkage of one small subunit with one large subunit of ribulose biphosphate carboxylase. For short periods of time (<1 h), the amount of P65 formed increased with the duration of the exposure to the UV radiation and with the energy of the radiation applied. Prolonged exposure to UV radiation (1-6 h) resulted in the formation of high-molecular-mass aggregates of ribulose biphosphate carboxylase. Formation of P65 was shown to depend on the native state of the protein, was stimulated by inhibitors of enzyme activity, and was inhibited by activators of enzyme activity. A UV-independent accumulation of P65 was also achieved by the in vitro incubation of plant crude extracts. However, the UV-dependent and the UV-independent formation of P65 seemed to occur by distinct molecular mechanisms. The UV-dependent accumulation of P65 was immunologically detected in all species examined, including Lemna minor, Arum italicum, Brassica oleracea, Triticum aestivum, Zea mays, Pisum sativum and Phaseolus vulgaris, suggesting that it may constitute a universal response to UV radiation, common to all photosynthetic tissues. (Author).

  6. Phytochrome control of gene expression in radish seedlings. II Far-red light mediated appearance of the ribulose 1,5-bisphosphate carboxylase and the mRNA for its small subunit

    International Nuclear Information System (INIS)

    The effect of far-red light on the appearance of the radish ribulose 1,5-bisphosphate carboxylase, its subunits and the mRNA for the small subunit has been studied. The immunological analysis of the accumulation of holoenzyme and small subunit showed that there was no pool of free small subunit either in the etiolated seedlings or in the far-red light illuminated seedlings. The precursor to the small subunit has been identified by immunoprecipitation of the in vitro translation products directed by poly(A)-containing RNA of radish cotyledons. The irradiation of radish cotyledons with far-red light led to the apparent increase of the level of the translatable mRNA for the small subunit. (author)

  7. Photocontrol of the expression of genes encoding chlorophyll a/b binding proteins and small subunit of ribulose-1,5-bisphosphate carboxylase in etiolated seedlings of Lycopersicon esculentum (L. ) and Nicotiana tabacum (L. )

    Energy Technology Data Exchange (ETDEWEB)

    Wehmeyer, B. (Univ. of Pennsylvania, Philadelphia (USA) Albert-Ludwigs-Universitaet, Freiburg (West Germany)); Cashmore, A.R. (Univ. of Pennsylvania, Philadelphia (USA)); Schaefer, E. (Albert-Ludwigs-Universitaet, Freiburg (West Germany))

    1990-07-01

    Phytochrome and the blue ultraviolet-A photoreceptor control light-induced expression of genes encoding the chlorophyll a/b binding protein of photosystem II and photosystem I and the genes for the small subunit of the ribulose-1,5-bisphosphate carboxylase in etiolated seedlings of Lycopersicon esculentum (tomato) and Nicotiana tabacum (tobacco). A high irradiance response also controls the induction of these genes. Genes encoding photosystem II- and I-associated chlorophyll a/b binding proteins both exhibit a transient rapid increase in expression in response to light pulse or to continuous irradiation. In contrast, genes encoding the small subunit exhibit a continuous increase in expression in response to light. These distinct expression characteristics are shown to reflect differences at the level of transcription.

  8. Crystallization and structure of a recombinant ribulose-1,5-bisphosphate carboxylase

    Science.gov (United States)

    Schneider, Gunter; Lindqvist, Ylva; Brändén, Carl-Ivar; Lorimer, George

    1988-07-01

    Ribulose-1,5-bisphosphate carboxylase/oxygenase is the key enzyme in photosynthetic carbon dioxide fixation and photorespiration. The dimeric carboxylase from the photosynthetic bacterium Rhodospirillum rubrum has been cloned and expressed in E. coli. The recombinant enzyme has been crystallized in a number of different crystal forms. The three-dimensional structure of the enzyme has been determined by X-ray crystallographic methods to 2.9Åresolution.

  9. Isolation of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase from Leaves

    Science.gov (United States)

    Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a multi-functional enzyme that catalyzes the fixation of CO2 and O2 in photosynthesis and photorespiration, respectively. As the rate-limiting step in photosynthesis, improving the catalytic properties of Rubisco has long been viewed as a...

  10. Isolation of ribulose-1,5-bisphosphate carboxylase/oxygenase from leaves.

    Science.gov (United States)

    Carmo-Silva, A Elizabete; Barta, Csengele; Salvucci, Michael E

    2011-01-01

    Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a multifunctional enzyme that catalyzes the fixation of CO2 and O2 in photosynthesis and photorespiration, respectively. As the rate-limiting step in photosynthesis, improving the catalytic properties of Rubisco has long been viewed as a viable strategy for increasing plant productivity. Advances in biotechnology have made this goal more attainable by making it possible to modify Rubisco in planta. To properly evaluate the properties of Rubisco, it is necessary to isolate the enzyme in pure form. This chapter describes procedures for rapid and efficient purification of Rubisco from leaves of several species. PMID:20960141

  11. Isolation, identification, and synthesis of 2-carboxyarabinitol 1-phosphate, a diurnal regulator of ribulase-bisphosphate carboxylase activity

    International Nuclear Information System (INIS)

    The diurnal change in activity of ribulose 1,5-bisphosphate (Rbu-1,5-P2) carboxylase [3-phospho-D-glycerate carboxy-lyase (dimerizing); EC 4.1.1.39] of leaves of Phaseolus vulgaris is regulated (in part) by mechanisms that control the level of an endogenous inhibitor that binds tightly to the activated (carbamoylated) form of Rbu-1,5-P2 carboxylase. This inhibitor was extracted from leaves and copurified with the Rbu-1,5-P2 carboxylase of the leaves. Further purification by ion-exchange chromatography, adsorption to purified Rbu-1,5-P2 carboxylase, barium precipitation, and HPLC separation yielded a phosphorylated compound that was a strong inhibitor of Rbu-1,5-P2 carboxylase. The compound was analyzed by GC/MS, 13C NMR, and 1H NMR and shown to be 2-carboxyarabinitol 1-phosphate [(2-C-phosphohydroxymethyl)-D-ribonic acid]. The structure of the isolated compound differs from the Rbu-1,5-P2 carboxylase transition-state analogue 2-carboxyarabinitol 1,5-bisphosphate only by the lack of the C-5 phosphate group. This difference results in a higher binding constant for the monophosphate compared with the bisphosphate. The less tightly bound compound acts in a light-dependent, reversible regulation of Rbu-1,5-P2 carboxylase activity in vivo

  12. Dark/light modulation of ribulose bisphosphate carboxylase activity in plants from different photosynthetic categories

    Energy Technology Data Exchange (ETDEWEB)

    Vu, J.C.V.; Allen, L.H. Jr.; Bowes, G.

    1984-11-01

    Ribulose bisphosphate carboxylase/oxygenase (RuBPCase) from several plants had substantially greater activity in extracts from light-exposed leaves than dark leaves, even when the extracts were incubated in vitro with saturating HCO/sub 3//sup -/ and Mg/sup 2 +/ concentrations. This occurred in Glycine max, Lycopersicon esculentum, Nicotiana tabacum, Panicum bisulcatum, and P. hylaeicum (C/sub 3/); P. maximum (C/sub 4/ phosphoenolpyruvate carboxykinase); P. milioides (C/sub 3//C/sub 4/); and Bromelia pinguin and Ananas comosus (Crassulacean acid metabolism). Little or no difference between light and dark leaf extracts of RuBPCase was observed in Triticum aestivum (C/sub 3/); P. miliaceum (C/sub 4/ NAD malic enzyme); Zea mays and Sorghum bicolor (C/sub 4/ NADP malic enzyme); Moricandia arvensis (C/sub 3//C/sub 4/); and Hydrilla verticillata (submersed aquatic macrophyte). It is concluded that, in many plants, especially Crassulacean acid metabolism and C/sub 3/ species, a large fraction of ribulose-1,5-bisphosphate carboxylase/oxygenase in the dark is in an inactivatable state that cannot respond to CO/sub 2/ and Mg/sup 2 +/ activation, but which can be converted to an activatable state upon exposure of the leaf to light. 16 references, 2 tables.

  13. Dark/Light Modulation of Ribulose Bisphosphate Carboxylase Activity in Plants from Different Photosynthetic Categories 1

    Science.gov (United States)

    Vu, J. Cu V.; Allen, Leon H.; Bowes, George

    1984-01-01

    Ribulose bisphosphate carboxylase/oxygenase (RuBPCase) from several plants had substantially greater activity in extracts from lightexposed leaves than dark leaves, even when the extracts were incubated in vitro with saturating HCO3− and Mg2+ concentrations. This occurred in Glycine max, Lycopersicon esculentum, Nicotiana tabacum, Panicum bisulcatum, and P. hylaeicum (C3); P. maximum (C4 phosphoenolpyruvate carboxykinase); P. milioides (C3/C4); and Bromelia pinguin and Ananas comosus (Crassulacean acid metabolism). Little or no difference between light and dark leaf extracts of RuBPCase was observed in Triticum aestivum (C3); P. miliaceum (C4 NAD malic enzyme); Zea mays and Sorghum bicolor (C4 NADP malic enzyme); Moricandia arvensis (C3/C4); and Hydrilla verticillata (submersed aquatic macrophyte). It is concluded that, in many plants, especially Crassulacean acid metabolism and C3 species, a large fraction of ribulose-1,5-bisphosphate carboxylase/oxygenase in the dark is in an inactivatable state that cannot respond to CO2 and Mg2+ activation, but which can be converted to an activatable state upon exposure of the leaf to light. PMID:16663937

  14. Dark/Light modulation of ribulose bisphosphate carboxylase activity in plants from different photosynthetic categories.

    Science.gov (United States)

    Vu, J C; Allen, L H; Bowes, G

    1984-11-01

    Ribulose bisphosphate carboxylase/oxygenase (RuBPCase) from several plants had substantially greater activity in extracts from lightexposed leaves than dark leaves, even when the extracts were incubated in vitro with saturating HCO(3) (-) and Mg(2+) concentrations. This occurred in Glycine max, Lycopersicon esculentum, Nicotiana tabacum, Panicum bisulcatum, and P. hylaeicum (C(3)); P. maximum (C(4) phosphoenolpyruvate carboxykinase); P. milioides (C(3)/C(4)); and Bromelia pinguin and Ananas comosus (Crassulacean acid metabolism). Little or no difference between light and dark leaf extracts of RuBPCase was observed in Triticum aestivum (C(3)); P. miliaceum (C(4) NAD malic enzyme); Zea mays and Sorghum bicolor (C(4) NADP malic enzyme); Moricandia arvensis (C(3)/C(4)); and Hydrilla verticillata (submersed aquatic macrophyte). It is concluded that, in many plants, especially Crassulacean acid metabolism and C(3) species, a large fraction of ribulose-1,5-bisphosphate carboxylase/oxygenase in the dark is in an inactivatable state that cannot respond to CO(2) and Mg(2+) activation, but which can be converted to an activatable state upon exposure of the leaf to light. PMID:16663937

  15. Ribulose-1,5-bisphosphate Carboxylase/Oxygenase content, assimilatory charge, and mesophyll conductance in leaves

    Science.gov (United States)

    Eichelmann; Laisk

    1999-01-01

    The content of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) (Et; EC 4.1.1.39) measured in different-aged leaves of sunflower (Helianthus annuus) and other plants grown under different light intensities, varied from 2 to 75 &mgr;mol active sites m-2. Mesophyll conductance (&mgr;) was measured under 1.5% O2, as well as postillumination CO2 uptake (assimilatory charge, a gas-exchange measure of the ribulose-1,5-bisphosphate pool). The dependence of &mgr; on Et saturated at Et = 30 &mgr;mol active sites m-2 and &mgr; = 11 mm s-1 in high-light-grown leaves. In low-light-grown leaves the dependence tended toward saturation at similar Et but reached a &mgr; of only 6 to 8 mm s-1. &mgr; was proportional to the assimilatory charge, with the proportionality constant (specific carboxylation efficiency) between 0.04 and 0.075 &mgr;M-1 s-1. Our data show that the saturation of the relationship between Et and &mgr; is caused by three limiting components: (a) the physical diffusion resistance (a minor limitation), (b) less than full activation of Rubisco (related to Rubisco activase and the slower diffusibility of Rubisco at high protein concentrations in the stroma), and (c) chloroplast metabolites, especially 3-phosphoglyceric acid and free inorganic phosphate, which control the reaction kinetics of ribulose-1,5-bisphosphate carboxylation by competitive binding to active sites. PMID:9880359

  16. Effect of low temperature stress on the transcription of ribulose-1, 5-bisphosphate carboxylase in rice seedlings

    International Nuclear Information System (INIS)

    The seeds of ri e (oryza sativa. cv. IR 8) were grown in the growth chambers having 14 hours day length and temperature 25 deg. C) under same photo period to observe low temperature effect. After 10 days the fourth leaf of each plant was excised and immediately killed in liquid nitrogen and then used for the extraction of total RNA by GTC method. This RNA was used to estimate the amount of mRNAs responsible for the transcription of lage subunit (LSU) and small subunit (|SSU) of the Ribulose-1, 5-bisphosphate carboxylase (RuPBC) by northern blot and dot blot hybridization using cDNA probes labeled with /sup 32/P. It was observed that amount of mRNAs (LSU and SSU mRNAs) or RuBPC decreased under low temperature stress indicating that transcription of RuBPC was affected. (author)

  17. Immunochemical localization of ribulose-1,5-bisphosphate carboxylase in the symbiont-containing gills of Solemya velum (Bivalvia : Mollusca)

    NARCIS (Netherlands)

    Cavanaugh, Colleen M.; Abbott, Marilyn S.; Veenhuis, Marten

    1988-01-01

    The distribution of the Calvin cycle enzyme ribulose-1,5-bisphosphate carboxylase (RbuP2Case; EC 4.1.1.39) was examined by using two immunological methods in tissues of Solemya velum, an Atlantic coast bivalve containing putative chemoautotrophic symbionts. Antibodies elicited by the purified large

  18. Identification of Interactions between Abscisic Acid and Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase.

    Directory of Open Access Journals (Sweden)

    Marek M Galka

    Full Text Available Abscisic acid ((+-ABA is a phytohormone involved in the modulation of developmental processes and stress responses in plants. A chemical proteomics approach using an ABA mimetic probe was combined with in vitro assays, isothermal titration calorimetry (ITC, x-ray crystallography and in silico modelling to identify putative (+-ABA binding-proteins in crude extracts of Arabidopsis thaliana. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco was identified as a putative ABA-binding protein. Radiolabelled-binding assays yielded a Kd of 47 nM for (+-ABA binding to spinach Rubisco, which was validated by ITC, and found to be similar to reported and experimentally derived values for the native ribulose-1,5-bisphosphate (RuBP substrate. Functionally, (+-ABA caused only weak inhibition of Rubisco catalytic activity (Ki of 2.1 mM, but more potent inhibition of Rubisco activation (Ki of ~ 130 μM. Comparative structural analysis of Rubisco in the presence of (+-ABA with RuBP in the active site revealed only a putative low occupancy (+-ABA binding site on the surface of the large subunit at a location distal from the active site. However, subtle distortions in electron density in the binding pocket and in silico docking support the possibility of a higher affinity (+-ABA binding site in the RuBP binding pocket. Overall we conclude that (+-ABA interacts with Rubisco. While the low occupancy (+-ABA binding site and weak non-competitive inhibition of catalysis may not be relevant, the high affinity site may allow ABA to act as a negative effector of Rubisco activation.

  19. Functional metagenomic selection of ribulose 1, 5-bisphosphate carboxylase/oxygenase from uncultivated bacteria.

    Science.gov (United States)

    Varaljay, Vanessa A; Satagopan, Sriram; North, Justin A; Witte, Brian; Dourado, Manuella N; Anantharaman, Karthik; Arbing, Mark A; McCann, Shelley Hoeft; Oremland, Ronald S; Banfield, Jillian F; Wrighton, Kelly C; Tabita, F Robert

    2016-04-01

    Ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) is a critical yet severely inefficient enzyme that catalyses the fixation of virtually all of the carbon found on Earth. Here, we report a functional metagenomic selection that recovers physiologically active RubisCO molecules directly from uncultivated and largely unknown members of natural microbial communities. Selection is based on CO2 -dependent growth in a host strain capable of expressing environmental deoxyribonucleic acid (DNA), precluding the need for pure cultures or screening of recombinant clones for enzymatic activity. Seventeen functional RubisCO-encoded sequences were selected using DNA extracted from soil and river autotrophic enrichments, a photosynthetic biofilm and a subsurface groundwater aquifer. Notably, three related form II RubisCOs were recovered which share high sequence similarity with metagenomic scaffolds from uncultivated members of the Gallionellaceae family. One of the Gallionellaceae RubisCOs was purified and shown to possess CO2 /O2 specificity typical of form II enzymes. X-ray crystallography determined that this enzyme is a hexamer, only the second form II multimer ever solved and the first RubisCO structure obtained from an uncultivated bacterium. Functional metagenomic selection leverages natural biological diversity and billions of years of evolution inherent in environmental communities, providing a new window into the discovery of CO2 -fixing enzymes not previously characterized. PMID:26617072

  20. Ribulose-1,5-bisphosphate carboxylase/oxygenase gene expression and diversity of Lake Erie planktonic microorganisms.

    OpenAIRE

    Xu, H.H.; Tabita, F R

    1996-01-01

    Carbon dioxide fixation is carried out primarily through the Calvin-Benson-Bassham reductive pentose phosphate cycle, in which ribulose-1, 5-bisphosphate carboxylase/oxygenase (RubisCO) is the key enzyme. The primary structure of the large subunit of form I RubisCO is well conserved; however, four distinct types, A, B, C, and D, may be distinguished, with types A and B and types C and D more closely related to one another. To better understand the environmental regulation of RubisCO in Lake E...

  1. Cloning and characterization of the gene product of the form II ribulose-1,5-bisphosphate carboxylase gene of Rhodopseudomonas sphaeroides.

    OpenAIRE

    Muller, E D; Chory, J; Kaplan, S

    1985-01-01

    We report the cloning and characterization of the gene product of the gene for the form II ribulose bisphosphate carboxylase from Rhodopseudomonas sphaeroides. We present evidence that the form II enzyme is encoded by a single gene in R. sphaeroides; however, this gene does hybridize to a second chromosomal locus.

  2. Status of the substrate binding sites of ribulose bisphosphate carboxylase as determined with 2-C-carboxyarabinitol 1,5-bisphosphate

    International Nuclear Information System (INIS)

    The properties of the tight and specific binding of 2-C-carboxy-D-arabinitol 1,5-bisphosphate (CABP), which occurs only to reaction sites of ribulose 1,5-bisphosphate carboxylase (Rubisco) that are activated by CO2 and Mg2+, were studied. With fully active purified spinach (Spinacia oleracea) Rubisco the rate of tight binding of [14C]CABP fit a multiple exponential rate equation with half of the sites binding with a rate constant of 40 per minute and the second half of the sites binding at 3.2 per minute. This suggests that after CABP binds to one site of a dimer of Rubisco large subunits, binding to the second site is considerably slower, indicating negative cooperativity as previously reported. The rate of CABP binding to partially activated Rubisco was complete within 2 to 5 minutes, with slower binding to inactive sites as they formed the carbamate and bound Mg2+. Addition of [14C]CABP and EDTA stopped binding of Mg2+ and allowed tight binding of the radiolabel only to sites which were CO2/Mg2+-activated at that moment. The rate of CO2 fixation was proportional to the CO2/Mg2+-activated sites. During light-dependent CO2 fixation with isolated spinach chloroplasts, the amount of carbamylation was proportional to Rubisco activity either initially upon lysis of the plastids or following total activation with Mg2+ and CO2. Lysis of chloroplasts in media with [14C]CABP plus EDTA estimated those carbamylated sites having Mg2+. The loss of Rubisco activation during illumination was partially due to the lack of Mg2+ to stabilize the carbamylated sites

  3. INFLUENCE OF NACL SALINITY ON THE ACTIVITY OF RIBULOSE 1, 5-BISPHOSPHATE CARBOXYLASE IN THREE LEAFY VEGETABLES

    Directory of Open Access Journals (Sweden)

    Anjali Ratnakar

    2015-06-01

    Full Text Available Salinity is one of the major factors which have an adverse effect on crop growth. Stress caused due to excessive accumulation of salts results in various physiological and biochemical changes in plants. Due to their nutritional value, leafy vegetables constitute an important part of our daily diet. In the present investigation, effect of NaCl salinity on the activity of ribulose 1,5–bisphosphate carboxylase in three leafy vegetables was studied. Seeds of leafy vegetables were sown in earthen pots and were subjected to different levels of saline water (NaCl treatment. Control plants were irrigated with tap water. Treatments started after the seedling emergence and continued till the plants were 45 day old. Mature leaves of these plants were harvested and used for the studies. The activity of ribulose 1,5-bisphosphate in all the three leafy vegetables was observed to decrease gradually with increase in the concentration of NaCl in the growth medium.

  4. Effects of Light and Elevated Atmospheric CO(2) on the Ribulose Bisphosphate Carboxylase Activity and Ribulose Bisphosphate Level of Soybean Leaves.

    Science.gov (United States)

    Vu, C V; Allen, L H; Bowes, G

    1983-11-01

    Soybean (Glycine max L. Merr. cv Bragg) was grown throughout its life cycle at 330, 450, and 800 microliters CO(2) per liter in outdoor controlled-environment chambers under solar irradiance. Leaf ribulose-1,5-bisphosphate carboxylase (RuBPCase) activities and ribulose-1,5-bisphosphate (RuBP) levels were measured at selected times after planting. Growth under the high CO(2) levels reduced the extractable RuBPCase activity by up to 22%, but increased the daytime RuBP levels by up to 20%.Diurnal measurements of RuBPCase (expressed in micromoles CO(2) per milligram chlorophyll per hour) showed that the enzyme values were low (230) when sampled before sunrise, even when activated in vitro with saturating HCO(3) (-) and Mg(2+), but increased to 590 during the day as the solar quantum irradiance (photosynthetically active radiation or PAR, in micromoles per square meter per second) rose to 600. The nonactivated RuBPCase values, which averaged 20% lower than the corresponding HCO(3) (-) and Mg(2+)-activated values, increased in a similar manner with increasing solar PAR. The per cent RuBPCase activation (the ratio of nonactivated to maximum-activated values) increased from 40% before dawn to 80% during the day. Leaf RuBP levels (expressed in nanomoles per milligram chlorophyll) were close to zero before sunrise but increased to a maximum of 220 as the solar PAR rose beyond 1200. In a chamber kept dark throughout the morning, leaf RuBPCase activities and RuBP levels remained at the predawn values. Upon removal of the cover at noon, the HCO(3) (-) and Mg(2+)-activated RuBPCase values and the RuBP levels rose to 465 and 122, respectively, after only 5 minutes of leaf exposure to solar PAR at 1500.These results indicate that, in soybean leaves, light may exert a regulatory effect on extractable RuBPCase in addition to the well-established activation by CO(2) and Mg(2+). PMID:16663291

  5. Effect of CO{sub 2} concentration on carbonic anhydrase and ribulose-1,5-bisphosphate carboxylase/oxygenase expression in pea

    Energy Technology Data Exchange (ETDEWEB)

    Majeau, N.; Coleman, J.R. [Univ. of Toronto, Ontario (Canada)

    1996-10-01

    The effect of external CO{sub 2} concentration on the expression of carbonic anhydrase (CA) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) was examined in pea (Pisum sativum cv Little Marvel) leaves. Enzyme activities and their transcript levels were reduced in plants grown at 1000 {mu}L/L CO{sub 2} compared with plants grown in ambient air. Growth at 160 {mu}L/L CO{sub 2} also appeared to reduce steady-state transcript levels for the rbcS, the gene encoding the small subunit of Rubisco, and for ca, the gene encoding CA; however, rbcS transcripts were reduced to a greater extent at this concentration. Rubisco activity was slightly lower in plants grown at 160 {mu}L/L CO{sub 2}, and CA activity was significantly higher than that observed in air-grown plants. Transfer of plants from 1000 {mu}L/L to air levels of CO{sub 2} resulted in a rapid increase in both ca and rbcS transcript abundance in fully expanded leaves, followed by an increase in enzyme activity. Plants transferred from air to high-CO{sub 2} concentrations appeared to modulate transcript abundance and enzyme activity less quickly. Foliar carbohydrate levels were also examined in plants grown continuously at high and ambient CO{sub 2}, and following changes in growth conditions that rapidly altered ca and rbcS transcript abundance and enzyme activities. 39 refs., 2 figs., 3 tabs.

  6. Ribulose Bisphosphate Carboxylase Activity in Anther-Derived Plants of Saintpaulia ionantha Wendl. Shag.

    Science.gov (United States)

    Bhaskaran, S; Smith, R H; Finer, J J

    1983-11-01

    Plants obtained from anther culture of the African violet, Saintpaulia ionantha Wendl. ;Shag' and vegetatively cloned copies of the parent anther donor plant were examined for their ploidy and ribulose-1,5-biphosphate carboxylase (RuBPcase) activity. The cloned parent plants were all diploid and did not vary much in their nuclear DNA, chlorophyll, and RuBPcase activity. Some of the anther-derived plants were similar to the parent plants while others were not. Different levels of ploidy were observed among the androgenetic plants. RuBPcase activities higher than that of the parent plants were found in some anther-derived plants. However, there was no direct correlation between ploidy and RuBPcase activity. Expression of nuclear genes from a single parent in the anther-derived plants and it's diploidization or plastid changes during early stages of microsporogenesis or androgenesis are suggested as possible reasons for the variations observed among them. This could be a useful technique to obtain physiological variants which could be agronomically desirable. PMID:16663273

  7. Decline of activity and quantity of ribulose bisphosphate carboxylase/oxygenase and net photosynthesis in ozone-treated potato foliage

    Energy Technology Data Exchange (ETDEWEB)

    Dann, M.S.; Pell, E.J. (Pennsylvania State Univ., University Park (USA))

    1989-09-01

    The effect of ozone (O{sub 3}) on ribulose bisphosphate carboxylase/oxygenase (Rubisco) activity and quantity and net photosynthesis in greenhouse-grown Solanum tuberosum L. cv Norland foliage was studied in relation to oxidant-induced premature senescence. Plants, 26 days old, were exposed to 0.06 to 0.08 microliters per liter O{sub 3} from 1,000 to 1,600 hours for 4 days in a controlled environment chamber. On day 5, plants were exposed to a 6-hour simulated inversion in which O{sub 3} peaked at 0.12 microliters per liter. Net photosynthesis declined in response to O{sub 3} but recovered to near control levels 3 days after the exposure ended. Rubisco activity and quantity in control potato foliage increased and then decreased during the 12-day interval of the study. In some experiments foliage studied was physiologically mature and Rubisco activity had peaked when O{sub 3} exposure commenced. In those cases, O{sub 3} accelerated the decline in Rubisco activity. When less mature foliage was treated with O{sub 3}, the leaves never achieved the maximal level of Rubisco activity observed in control foliage and also exhibited more rapid decline in initial and total activity. Percent activation of Rubisco (initial/total activity) was not affected significantly by treatment. Quantity of Rubisco decreased in concert with activity. The reduction in the quantity of Rubisco, an important foliage storage protein, could contribute to premature senescence associated with toxicity of this air pollutant.

  8. Simple determination of the CO2/O2 specificity of Ribulose-1,5-bisphosphate carboxylase/oxygenase by the specific radioactivity of [14C] glycerate 3-phosphate

    International Nuclear Information System (INIS)

    A new method is presented for measurement of the CO2/O2 specificity factor of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). The [14C]3-phosphoglycerate (PGA) from the Rubisco carboxylase reaction and its dilution by the Rubisco oxygenase reaction was monitored by directly measuring the specific radioactivity of PGA. 14CO2 fixation with Rubisco occurred under two reaction conditions: carboxylase with oxygenase with 40 micromolar CO2 in O2-saturated water and carboxylase only with 160 micromolar CO2 under N2. Detection of the specific radioactivity used the amount of PGA as obtained from the peak area, which was determined by pulsed amperometry following separation by high-performance anion exchange chromatography and the radioactive counts of the [14C]PGA in the same peak. The specificity factor of Rubisco from spinach (Spinacia oleracea L.) (93 ± 4), from the green alga Chlamydomonas reinhardtii (66 ± 1), and from the photosynthetic bacterium Rhodospirillum rubrum (13) were comparable with the published values measured by different methods

  9. Dithiothreitol decreases the thermal stability and unfolding cooperativity of ribulose-1, 5-bisphosphate carboxylase/oxygenase

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Plant rubisco consists of eight large subunits (55 kD) encoded by chloroplast gene and eight small subunits (15 kD) encoded by nuclear gene. There are abundant cysteine residues that do not form disulfide bonds in native rubisco. Differential scanning calorimetry has been used to study some plant rubisco and suggested an irreversible two-state denaturation due to the high cooperativity in subunits. By comparing the data from circular dichroism, fluorescence, differential scanning calorimetry, SDS electrophoresis, and activity assays in the absence or presence of DTT, we suggest that the formation of disulfide bonds in subunits during the early thermal unfolding may increase the thermal stability and the thermal unfolding cooperativity of rubisco.

  10. Characterization of ribulose-1, 5-bisphosphate carboxylase/oxygenase and transcriptional analysis of its related genes in Saccharina japonica (Laminariales, Phaeophyta)

    Science.gov (United States)

    Shao, Zhanru; Liu, Fuli; Li, Qiuying; Yao, Jianting; Duan, Delin

    2014-03-01

    Saccharina japonica is a common macroalga in sublittoral communities of cold seawater environments, and consequently may have highly efficient ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) activity for carbon assimilation. In our study, we cloned the full-length Rubisco gene from S. japonica ( SJ-rbc). It contained an open reading frame for a large subunit gene ( SJ — rbcL) of 1 467 bp, a small subunit gene ( SJ-rbcS) of 420 bp, and a SJ-rbcL/S intergenic spacer of 269 bp. The deduced peptides of SJ-rbcL and SJ-rbcS were 488 and 139 amino acids with theoretical molecular weights and isoelectric points of 53.97 kDa, 5.81 and 15.84 kDa, 4.71, respectively. After induction with 1 mmol/L isopropyl- β-D-thiogalactopyranoside for 5 h and purification by Ni2+ affinity chromatography, electrophoresis and western blot detection demonstrated successful expression of the 55 kDa SJ-rbcL protein. Real-time quantitative PCR showed that the mRNA levels of SJ-rbcL in gametophytes increased when transferred into normal growth conditions and exhibited diurnal variations: increased expression during the day but suppressed expression at night. This observation implied that Rubisco played a role in normal gametophytic growth and development. In juvenile sporophytes, mRNA levels of SJ-rbcL, carbonic anhydrase, Calvin-Benson-Bassham cycle-related enzyme, and chloroplast light-harvesting protein were remarkably increased under continuous light irradiance. Similarly, expression of these genes was up-regulated under blue light irradiance at 350 μmol/(m2·s). Our results indicate that long-term white light and short-term blue light irradiance enhances juvenile sporophytic growth by synergistic effects of various photosynthetic elements.

  11. Mild water stress effects on carbon-reduction-cycle intermediates, ribulose bisphosphate carboxylase activity, and spatial homogeneity of photosynthesis in intact leaves

    International Nuclear Information System (INIS)

    We have examined the effect of mild water stress on photosynthetic chloroplast reactions of intact Phaseolus vulgaris leaves by measuring two parameters of ribulose bisphosphate (RuBP) carboxylase activity and the pool sizes of RuBP, 3-phosphoglycerate (PGA), triose phosphates, hexose monophosphates, and ATP. We also tested for patchy stomatal closure by feeding 14CO2. The kcat of RuBP carboxylase (moles CO2 fixed per mole enzyme per second) which could be measured after incubating the enzyme with CO2 and Mg2+ was unchanged by water stress. The ratio of activity before and after incubation with CO2 and Mg2+ (the carbamylation state) was slightly reduced by severe stress but not by mild stress. Likewise, the concentration of RuBP was slightly reduced by severe stress but not by mild stress. The concentration of PGA was markedly reduced by both mild and severe water stress. The concentration of triose phosphates did not decline as much as PGA. We found that photosynthesis in water stressed leaves occurred in patches. The patchiness of photosynthesis during water stress may lead to an underestimation of the effect of stomatal closure. We conclude that reductions in whole leaf photosynthesis caused by mild water stress are primarily the result of stomatal closure and that there is no indication of damage to chloroplast reactions

  12. Partitioning of Nitrogen among Ribulose-1,5-bisphosphate Carboxylase/Oxygenase, Phosphoenolpyruvate Carboxylase, and Pyruvate Orthophosphate Dikinase as Related to Biomass Productivity in Maize Seedlings.

    Science.gov (United States)

    Sugiyama, T; Mizuno, M; Hayashi, M

    1984-07-01

    Maize (Zea mays L. cv Golden Cross Bantam T51) seedlings were grown under full sunlight or 50% sunlight in a temperature-controlled glasshouse at the temperatures of near optimum (30/25 degrees C) and suboptimum (17/13 degrees C) with seven levels of nitrate-N (0.4 to 12 millimolars). The contents of phosphoenolpyruvate carboxylase (PEPC), pyruvate orthophosphate dikinase (PPD), and ribulose-1,5-P(2) carboxylase/oxygenase (RuBisCO) were immunochemically determined for each treatment with rabbit antibodies raised against the respective maize leaf proteins (anti-PEPC and anti-PPD) or spinach leaf protein (anti-RuBisCO). The content of each enzymic protein increased with increasing N and raised under reduced temperature. The positive effect of light intensity on their contents was evident only at near optimal temperature. The relative increase in PEPC and PPD content with increasing N was significantly greater than that of RuBisCO irrespective of growth conditions. These enzymic proteins comprised about 8, 6, and 35% of total soluble protein, respectively, at near optimal growth condition. In contrast to significant increase in the proportion of soluble protein allocated to PEPC and PPD seen under certain conditions, the proportion allocated to RuBisCO decreased reciprocally with an increased biomass yield by N supply.These results indicated that the levels of PEPC and PPD parallel to maize biomass more tightly than that of RuBisCO at least under near optimal growth condition. PMID:16663684

  13. Determining Photosynthetic Parameters from Leaf CO2 Exchange and Chlorophyll Fluorescence (Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Specificity Factor, Dark Respiration in the Light, Excitation Distribution between Photosystems, Alternative Electron Transport Rate, and Mesophyll Diffusion Resistance.

    Science.gov (United States)

    Laisk, A.; Loreto, F.

    1996-03-01

    Using simultaneous measurements of leaf gas exchange and chlorophyll fluorescence, we determined the excitation partitioning to photosystem II (PSII), the CO2/O2 specificity of ribulose-1,5-bisphosphate carboxylase/oxygenase, the dark respiration in the light, and the alternative electron transport rate to acceptors other than bisphosphoglycerate, and the transport resistance for CO2 in the mesophyll cells for individual leaves of herbaceous and tree species. The specificity of ribulose-1,5-bisphosphate carboxylase/oxygenase for CO2 was determined from the slope of the O2 dependence of the CO2 compensation point between 1.5 and 21% O2. Its value, on the basis of dissolved CO2 and O2 concentrations at 25.5[deg]C, varied between 86 and 89. Dark respiration in the light, estimated from the difference between the CO2 compensation point and the CO2 photocompensation point, was about 20 to 50% of the respiration rate in the dark. The excitation distribution to PSII was estimated from the extrapolation of the dependence of the PSII quantum yield on F/Fm to F = 0, where F is steady-state and Fm is pulse-satuarated fluorescence, and varied between 0.45 and 0.6. The alternative electron transport rate was found as the difference between the electron transport rates calculated from fluorescence and from gas exchange, and at low CO2 concentrations and 10 to 21% O2, it was 25 to 30% of the maximum electron transport. The calculated mesophyll diffusion resistance accounted for about 20 to 30% of the total mesophyll resistance, which also includes carboxylation resistance. Whole-leaf photosynthesis is limited by gas phase, mesophyll diffusion, and carboxylation resistances in nearly the same proportion in both herbaceous species and trees. PMID:12226229

  14. The nature and alternate rates of the ribulose 1,5-bisphosphate (RuBP) carboxylase/oxygenase (Rubisco) oxygenation intermediate

    Energy Technology Data Exchange (ETDEWEB)

    Harpel, M.R.; Chen, Yuh-Ru; Hartman, F.C.

    1995-12-31

    Mutant ribulose 1,5-bisphosphate (RuBP) were employed to investigate the partitioning of carbon flow between photosynthesis or photorespiration. Previous functional and structural studies implicate active site Lys329 and Glu48 or R. rubrum RuBp in promoting addition of CO2 to the RuBP-enediol. Two novel O2-dependent side products generated by the K329A and E49Q mutants provided insight into RuBP oxygenase intermediate and roles of Lys329 and Glu48 in oxygenation.

  15. Influence of oxygen on the level of ribulose-1.5-bisphosphate carboxylase/oxygenase (Rubisco): only a substrate or also an inhibitor?

    International Nuclear Information System (INIS)

    Influence of oxygen upon the carboxylation of ribulose-1,5-bisphosphate (RuBP) in wheat (Triticum aestivum) leaves was studied at the steady state photosynthesis and at sharp transfers to a higher or to a lower 02 concentration. In leaves exhibiting a 26% Warburg effect at the steady state photosynthesis in the saturating light, no inhibition of carboxylation by oxygen was observed when the rate was measured by means of labelling during a 0.2-s exposure. The radioactivity of the extracts and of the residue was measured by means of a scintillation counter. The RuBP content of leaves was estimated by the post illumination tracer uptake at the exposure to 10 000 ppm 14CO2. Linearity of the measured carboxylation rate to the C02 concentration over the range from 0 to 100% showed that the uptake represented a bimolecular reaction. The rate estimated by the method applied was a characteristic of the primary binding of C02, a partial reaction of carboxylation, not of the total turnover of the catalytic site. Independence of this partial reaction of O2 indicated that oxygen did not inhibit carboxylation, neither did it cause any significant reduction in the pool of the immediate C02 acceptor, the Rubisco-RuBP complex, despite its role as an alternative substrate. 3 figs., 2 tabs.; 13 refs

  16. Effects of manganese-excess on CO2 assimilation, ribulose-1,5-bisphosphate carboxylase/oxygenase, carbohydrates and photosynthetic electron transport of leaves, and antioxidant systems of leaves and roots in Citrus grandis seedlings

    Directory of Open Access Journals (Sweden)

    Tang Ning

    2010-03-01

    Full Text Available Abstract Background Very little is known about the effects of manganese (Mn-excess on citrus photosynthesis and antioxidant systems. Seedlings of sour pummelo (Citrus grandis were irrigated for 17 weeks with nutrient solution containing 2 μM (control or 500 μM (excess MnSO4. The objective of this study were to understand the mechanisms by which Mn-excess leads to a decrease in CO2 assimilation and to test the hypothesis that Mn-induced changes in antioxidant systems differ between roots and leaves. Results Mn-excess decreased CO2 assimilation and stomatal conductance, increased intercellular CO2 concentration, but did not affect chlorophyll (Chl level. Both initial and total ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco activity in Mn-excess leaves decreased to a lesser extent than CO2 assimilation. Contents of glucose, fructose, starch and total nonstructural carbohydrates did not differ between Mn-excess leaves and controls, while sucrose content was higher in the former. Chl a fluorescence (OJIP transients from Mn-excess leaves showed increased O-step and decreased P-step, accompanied by positive L- and K-bands. Mn-excess decreased maximum quantum yield of primary photochemistry (Fv/Fm and total performance index (PItot,abs, but increased relative variable fluorescence at I-steps (VI and energy dissipation. On a protein basis, Mn-excess leaves displayed higher activities of monodehydroascorbate reductase (MDAR, glutathione reductase (GR, superoxide dismutase (SOD, catalase (CAT and guaiacol peroxidase (GPX and contents of antioxidants, similar ascorbate peroxidase (APX activities and lower dehydroascorbate reductase (DHAR activities; while Mn-excess roots had similar or lower activities of antioxidant enzymes and contents of antioxidants. Mn-excess did not affect malondialdehyde (MDA content of roots and leaves. Conclusions Mn-excess impaired the whole photosynthetic electron transport chain from the donor side of photosystem II

  17. Pyruvate carboxylase is critical for non-small-cell lung cancer proliferation.

    Science.gov (United States)

    Sellers, Katherine; Fox, Matthew P; Bousamra, Michael; Slone, Stephen P; Higashi, Richard M; Miller, Donald M; Wang, Yali; Yan, Jun; Yuneva, Mariia O; Deshpande, Rahul; Lane, Andrew N; Fan, Teresa W-M

    2015-02-01

    Anabolic biosynthesis requires precursors supplied by the Krebs cycle, which in turn requires anaplerosis to replenish precursor intermediates. The major anaplerotic sources are pyruvate and glutamine, which require the activity of pyruvate carboxylase (PC) and glutaminase 1 (GLS1), respectively. Due to their rapid proliferation, cancer cells have increased anabolic and energy demands; however, different cancer cell types exhibit differential requirements for PC- and GLS-mediated pathways for anaplerosis and cell proliferation. Here, we infused patients with early-stage non-small-cell lung cancer (NSCLC) with uniformly 13C-labeled glucose before tissue resection and determined that the cancerous tissues in these patients had enhanced PC activity. Freshly resected paired lung tissue slices cultured in 13C6-glucose or 13C5,15N2-glutamine tracers confirmed selective activation of PC over GLS in NSCLC. Compared with noncancerous tissues, PC expression was greatly enhanced in cancerous tissues, whereas GLS1 expression showed no trend. Moreover, immunohistochemical analysis of paired lung tissues showed PC overexpression in cancer cells rather than in stromal cells of tumor tissues. PC knockdown induced multinucleation, decreased cell proliferation and colony formation in human NSCLC cells, and reduced tumor growth in a mouse xenograft model. Growth inhibition was accompanied by perturbed Krebs cycle activity, inhibition of lipid and nucleotide biosynthesis, and altered glutathione homeostasis. These findings indicate that PC-mediated anaplerosis in early-stage NSCLC is required for tumor survival and proliferation. PMID:25607840

  18. Serine 363 of a Hydrophobic Region of Archaeal Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase from Archaeoglobus fulgidus and Thermococcus kodakaraensis Affects CO2/O2 Substrate Specificity and Oxygen Sensitivity.

    Directory of Open Access Journals (Sweden)

    Nathan E Kreel

    Full Text Available Archaeal ribulose 1, 5-bisphospate carboxylase/oxygenase (RubisCO is differentiated from other RubisCO enzymes and is classified as a form III enzyme, as opposed to the form I and form II RubisCOs typical of chemoautotrophic bacteria and prokaryotic and eukaryotic phototrophs. The form III enzyme from archaea is particularly interesting as several of these proteins exhibit unusual and reversible sensitivity to molecular oxygen, including the enzyme from Archaeoglobus fulgidus. Previous studies with A. fulgidus RbcL2 had shown the importance of Met-295 in oxygen sensitivity and pointed towards the potential significance of another residue (Ser-363 found in a hydrophobic pocket that is conserved in all RubisCO proteins. In the current study, further structure/function studies have been performed focusing on Ser-363 of A. fulgidus RbcL2; various changes in this and other residues of the hydrophobic pocket point to and definitively establish the importance of Ser-363 with respect to interactions with oxygen. In addition, previous findings had indicated discrepant CO2/O2 specificity determinations of the Thermococcus kodakaraensis RubisCO, a close homolog of A. fulgidus RbcL2. It is shown here that the T. kodakaraensis enzyme exhibits a similar substrate specificity as the A. fulgidus enzyme and is also oxygen sensitive, with equivalent residues involved in oxygen interactions.

  19. Mechanism and Significance of Post-Translational Modifications in the Large (LS) and Small (SS) Subunits of Ribulose-1,5 Bisphosphate Carboxylase/Oxygenase

    Energy Technology Data Exchange (ETDEWEB)

    Houtz, Robert, L.

    2012-11-09

    This project focused on a molecular and biochemical characterization of the protein methyltransferases responsible for methylation of the LS and SS in Rubisco, and the associated functional consequences accompanying these modifications. Our results provided some of the most informative structural and mechanistic understandings of SET domain protein methyltransferases. These results also positioned us to provide the first unambiguous assignment of the kinetic reaction mechanism for SET-domain protein methyltransferases, and to design and engineer an alternative substrate for Rubisco LSMT, enabling substrate specificity and functional significance studies. We demonstrated that the minimal substrate recognized by Rubisco LSMT is free lysine as well as monomethyllysine, an observation corroborated both by structural analyses as well as enzymatic activity and subsequent product distribution analyses. Ternary complexes between Rubisco LSMT and free lysine compared to complexes with monomethyllysine demonstrated that the structural basis for multiple methyl group additions is a consequence of hydrogen-bond driven spatial shifts in the amino group of Lys-14, which maintains the direct in-line geometry necessary for SN2 nucleophilic attack. The structural observations are also consistent with the previous proposal that the multiplicity of methyl group additions takes place through a processive mechanism, with successive methyl group additions to an enzyme protein complex which does not disassociate prior to the formation of trimethyllysine. This mechanism has important implications, since the regulation of gene expression by SET domain histone methyltransferases is not only dependent on site-specific lysine methylation, but also the degree of methylation. We examined the kinetic reaction mechanism for three different types of SET domain protein methyltransferases, each under conditions supporting mono-, di-, or trimethyllysine formation corroborated by product analyses. Additionally, the tight initial binding of Rubisco LSMT to Rubisco also allowed us to design a novel immobilized complex between Rubisco and Rubisco LSMT, which allowed for an unambiguous demonstration of the requirement for trimethyllysine formation prior to disassociation of the Rubisco LSMT:Rubisco complex, and therefore proof of the processive mechanism for methyl group transfer. These kinetic studies also demonstrated that an important factor has been overlooked in all kinetic analyses of SET domain protein methyltransferases reported to date. This factor is the influence of the low turnover number for SET domain protein methyltransferases and how, relative to the time-frame of kinetic enzyme assays, this can generate changes in kinetic profiles shifting reciprocal plot patterns from random/ordered bi-bi to the real kinetic reaction mechanism plots of ping-pong. Although the ternary complexes of Rubisco LSMT with S-Adenosylhomocysteine and lysine and monomethyllysine were informative in regard to reaction mechanism, they were not helpful in identifying the mechanism used by Rubisco LSMT for determining substrate specificity. We were unsuccessful at obtaining ternary complexes of Rubisco LSMT with bound synthetic polypeptide substrates, as has been reported for several histone methyltransferases. However, we were able to model a polypeptide sequence corresponding to the N-terminal region of the LS of Rubisco into the apparent substrate binding cleft in Rubisco LSMT. Knowledge of the determinants of polypeptide substrate specificity are important for identifying possible alternate substrates, as well as the possibility of generating more desirable substrates amenable to site-directed mutagenesis experiments unlike Rubisco. We determined that Rubisco LSMT is capable of methylating synthetic polypeptide mimics of the N-terminal region of the LS, both free as well as conjugated to keyhole limpet hemacyanin, but with considerable less efficiency than intact holoenzyme.

  20. Protein (Cyanobacteria): 246918 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available 5-bisphosphate carboxylase small subunit Moorea producens 3L MKTLPKERRYETLSYLPPLSDAQIMRQVEYILAEGYIPAIEFNESSEPEIYYWTLWKLPLFKATSPKDVLAEVDECRSEYRDCYIRVVGFDNVKQCQVLSFIIHKPNEGVSRSRW ...

  1. Molecular evolution of urea amidolyase and urea carboxylase in fungi

    Directory of Open Access Journals (Sweden)

    Harris Steven D

    2011-03-01

    Full Text Available Abstract Background Urea amidolyase breaks down urea into ammonia and carbon dioxide in a two-step process, while another enzyme, urease, does this in a one step-process. Urea amidolyase has been found only in some fungal species among eukaryotes. It contains two major domains: the amidase and urea carboxylase domains. A shorter form of urea amidolyase is known as urea carboxylase and has no amidase domain. Eukaryotic urea carboxylase has been found only in several fungal species and green algae. In order to elucidate the evolutionary origin of urea amidolyase and urea carboxylase, we studied the distribution of urea amidolyase, urea carboxylase, as well as other proteins including urease, across kingdoms. Results Among the 64 fungal species we examined, only those in two Ascomycota classes (Sordariomycetes and Saccharomycetes had the urea amidolyase sequences. Urea carboxylase was found in many but not all of the species in the phylum Basidiomycota and in the subphylum Pezizomycotina (phylum Ascomycota. It was completely absent from the class Saccharomycetes (phylum Ascomycota; subphylum Saccharomycotina. Four Sordariomycetes species we examined had both the urea carboxylase and the urea amidolyase sequences. Phylogenetic analysis showed that these two enzymes appeared to have gone through independent evolution since their bacterial origin. The amidase domain and the urea carboxylase domain sequences from fungal urea amidolyases clustered strongly together with the amidase and urea carboxylase sequences, respectively, from a small number of beta- and gammaproteobacteria. On the other hand, fungal urea carboxylase proteins clustered together with another copy of urea carboxylases distributed broadly among bacteria. The urease proteins were found in all the fungal species examined except for those of the subphylum Saccharomycotina. Conclusions We conclude that the urea amidolyase genes currently found only in fungi are the results of a horizontal

  2. Fructose-2,6-bisphosphate

    DEFF Research Database (Denmark)

    Nielsen, Tom Hamborg; Rung, Jesper Henrik; Villadsen, Dorthe

    2004-01-01

    Fructose-2,6-bisphosphate (Fru-2,6-P2) regulates key reactions of the primary carbohydrate metabolism in all eukaryotes. In plants, Fru-2,6-P2 coordinates the photosynthetic carbon flux into sucrose and starch biosynthesis. The use of transgenic plants has allowed the regulatory models to be tested...

  3. Demonstration of a functional requirement for the carbamate nitrogen of ribulosebisphosphate carboxylase/oxygenase by chemical rescue

    International Nuclear Information System (INIS)

    Ribulosebisphosphate carboxylase/oxygenase is reversibly activated by the reaction of Co2 with a specific lysyl residue to form a carbamate that coordinates an essential Mg2+ cation. Surprisingly, the Lys191→Cys mutant protein, in the presence of Co2 and Mg2+ exhibits tight binding of the reaction intermediate analogue 2-carboxyarabinitol bisphosphate a property normally equated with effective coordination of the Mg2+ by the carbamate. Catalytic ineptness of the Cys191 mutant protein, despite its ability to coordinate Mg2+ properly, might be due to the absence of the carbamate nitrogen. To investigate this possibility, the authors have evaluated the ability of exogenous amines to restore catalytic activity to the mutant protein. Significantly, the Cys191 protein manifests ribulose bisphosphate dependent fixation of 14CO2 when incubated with aminomethanewsulfonate but not ethanesulfonate. This novel activity reflects a Km value for ribulose bisphosphate which is not markedly perturbed relative to wild-type enzyme, a Km for Mg2+ which is in fact decreased 10-fold, and rate saturation with respect to aminomethanesulfonate. Chromatographic and spectrophotometric analyses reveal the product of CO2 fixation to be D-3-phosphoglycerate while turnover of [1-3H]ribulose bisphosphate into [3H]phosphoglycolate confirms oxygenase activity. The authors conclude that aminomethanesulfonate restored ribulosebisphosphate carboxylase/oxygenase activities to the Cys191 mutant protein by providing a nitrogenous function which satisfies a catalytic demand normally met by the carbamate nitrogen of Lys191

  4. AcEST: DK947508 [AcEST

    Lifescience Database Archive (English)

    Full Text Available e bisphosphate carboxylase small c... 111 2e-23 tr|A6MZQ8|A6MZQ8_ORYSI Ribulose bisphosphate carboxylase sma...ll c... 111 2e-23 tr|A3CGJ9|A3CGJ9_ORYSJ Ribulose bisphosphate carboxylase small c... 111 2e-23 tr|A2ZJQ0|A2

  5. Active site histidine in spinach ribulosebisphosphate carboxylase/oxygenase modified by diethyl pyrocarbonate

    International Nuclear Information System (INIS)

    [3H] Diethyl pyrocarbonate was synthesized from [3H] ethanol prepared by the reduction of acetaldehyde by NaB3H4. Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) from spinach was inactivated with this reagent at pH 7.0 the presence of 20 mM Mg2+, and tryptic peptides that contained modified histidine residues were isolated by reverse-phase high-performance liquid chromatography. Labeling of the enzyme was conducted in the presence and absence of the competitive inhibitor sedoheptulose 1,7-bisphosphate. The amount of one peptide that was heavily labeled in the absence of this compound was reduced 10-fold in its presence. The labeled residue was histidine-298. This result, in combination with earlier experiments, suggests that His-298 in spinach RuBisCO is located in the active site domain and is essential to enzyme activity. This region of the primary structure is strongly conserved in seven other ribulosebisphosphate carboxylases from divergent sources

  6. Chemical Genetics of Acetyl-CoA Carboxylases

    Directory of Open Access Journals (Sweden)

    Xuyu Zu

    2013-01-01

    Full Text Available Chemical genetic studies on acetyl-CoA carboxylases (ACCs, rate-limiting enzymes in long chain fatty acid biosynthesis, have greatly advanced the understanding of their biochemistry and molecular biology and promoted the use of ACCs as targets for herbicides in agriculture and for development of drugs for diabetes, obesity and cancers. In mammals, ACCs have both biotin carboxylase (BC and carboxyltransferase (CT activity, catalyzing carboxylation of acetyl-CoA to malonyl-CoA. Several classes of small chemicals modulate ACC activity, including cellular metabolites, natural compounds, and chemically synthesized products. This article reviews chemical genetic studies of ACCs and the use of ACCs for targeted therapy of cancers.

  7. Comparative modeling and molecular dynamics suggest high carboxylase activity of the Cyanobium sp. CACIAM14 RbcL protein.

    Science.gov (United States)

    Siqueira, Andrei Santos; Lima, Alex Ranieri Jerônimo; Dall'Agnol, Leonardo Teixeira; de Azevedo, Juliana Simão Nina; da Silva Gonçalves Vianez, João Lídio; Gonçalves, Evonnildo Costa

    2016-03-01

    Rubisco catalyzes the first step reaction in the carbon fixation pathway, bonding atmospheric CO2/O2 to ribulose 1,5-bisphosphate; it is therefore considered one of the most important enzymes in the biosphere. Genetic modifications to increase the carboxylase activity of rubisco are a subject of great interest to agronomy and biotechnology, since this could increase the productivity of biomass in plants, algae and cyanobacteria and give better yields in crops and biofuel production. Thus, the aim of this study was to characterize in silico the catalytic domain of the rubisco large subunit (rbcL gene) of Cyanobium sp. CACIAM14, and identify target sites to improve enzyme affinity for ribulose 1,5-bisphosphate. A three-dimensional model was built using MODELLER 9.14, molecular dynamics was used to generate a 100 ns trajectory by AMBER12, and the binding free energy was calculated using MM-PBSA, MM-GBSA and SIE methods with alanine scanning. The model obtained showed characteristics of form-I rubisco, with 15 beta sheets and 19 alpha helices, and maintained the highly conserved catalytic site encompassing residues Lys175, Lys177, Lys201, Asp203, and Glu204. The binding free energy of the enzyme-substrate complexation of Cyanobium sp. CACIAM14 showed values around -10 kcal mol(-1) using the SIE method. The most important residues for the interaction with ribulose 1,5-bisphosphate were Arg295 followed by Lys334. The generated model was successfully validated, remaining stable during the whole simulation, and demonstrated characteristics of enzymes with high carboxylase activity. The binding analysis revealed candidates for directed mutagenesis sites to improve rubisco's affinity. PMID:26936271

  8. Design of small molecule inhibitors of acetyl-CoA carboxylase 1 and 2 showing reduction of hepatic malonyl-CoA levels in vivo in obese Zucker rats.

    Science.gov (United States)

    Bengtsson, Christoffer; Blaho, Stefan; Saitton, David Blomberg; Brickmann, Kay; Broddefalk, Johan; Davidsson, Ojvind; Drmota, Tomas; Folmer, Rutger; Hallberg, Kenth; Hallén, Stefan; Hovland, Ragnar; Isin, Emre; Johannesson, Petra; Kull, Bengt; Larsson, Lars-Olof; Löfgren, Lars; Nilsson, Kristina E; Noeske, Tobias; Oakes, Nick; Plowright, Alleyn T; Schnecke, Volker; Ståhlberg, Pernilla; Sörme, Pernilla; Wan, Hong; Wellner, Eric; Oster, Linda

    2011-05-15

    Inhibition of acetyl-CoA carboxylases has the potential for modulating long chain fatty acid biosynthesis and mitochondrial fatty acid oxidation. Hybridization of weak inhibitors of ACC2 provided a novel, moderately potent but lipophilic series. Optimization led to compounds 33 and 37, which exhibit potent inhibition of human ACC2, 10-fold selectivity over inhibition of human ACC1, good physical and in vitro ADME properties and good bioavailability. X-ray crystallography has shown this series binding in the CT-domain of ACC2 and revealed two key hydrogen bonding interactions. Both 33 and 37 lower levels of hepatic malonyl-CoA in vivo in obese Zucker rats. PMID:21515056

  9. Fructose 2,6-bisphosphate in rat erythrocytes. Inhibition of fructose 2,6-bisphosphate synthesis and measurement by glycerate 2,3-bisphosphate.

    OpenAIRE

    Sobrino, F; Rider, Mark; Gualberto, A; Hue, Louis

    1987-01-01

    The concentration of fructose 2,6-bisphosphate found in freshly isolated erythrocytes was below the limit of detection (20 pmol/ml of packed cells). However, it increased to about 250 pmol/ml of cells when erythrocytes were incubated with glucose at pH 6.9, but not at pH 7.4 or 8.2. This could be explained by variations in the content of glycerate 2,3-bisphosphate, which was found to inhibit 6-phosphofructo-2-kinase, the enzyme responsible for fructose 2,6-bisphosphate synthesis. Glycerate 2,...

  10. Structure of fructose bisphosphate aldolase from Bartonella henselae bound to fructose 1,6-bisphosphate

    International Nuclear Information System (INIS)

    While other aldolases crystallize readily in the apo form, diffraction-quality crystals of B. henselae aldolase could only be obtained in the presence of the native substrate. The quaternary structure is tetrameric, as is typical of aldolases. Fructose bisphosphate aldolase (FBPA) enzymes have been found in a broad range of eukaryotic and prokaryotic organisms. FBPA catalyses the cleavage of fructose 1,6-bisphosphate into glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. The SSGCID has reported several FBPA structures from pathogenic sources, including the bacterium Brucella melitensis and the protozoan Babesia bovis. Bioinformatic analysis of the Bartonella henselae genome revealed an FBPA homolog. The B. henselae FBPA enzyme was recombinantly expressed and purified for X-ray crystallographic studies. The purified enzyme crystallized in the apo form but failed to diffract; however, well diffracting crystals could be obtained by cocrystallization in the presence of the native substrate fructose 1,6-bisphosphate. A data set to 2.35 Å resolution was collected from a single crystal at 100 K. The crystal belonged to the orthorhombic space group P212121, with unit-cell parameters a = 72.39, b = 127.71, c = 157.63 Å. The structure was refined to a final free R factor of 22.2%. The structure shares the typical barrel tertiary structure and tetrameric quaternary structure reported for previous FBPA structures and exhibits the same Schiff base in the active site

  11. Electron spin resonance studies of ribulosebisphosphate carboxylase: identification of activator cation ligands.

    Science.gov (United States)

    Miziorko, H M; Sealy, R C

    1984-01-31

    Ribulosebisphosphate carboxylase (RuBP carboxylase)forms a stable model complex containing stoichiometric amounts of enzyme sites, activator C0(2), divalent activator cation, and the transition-state analogue carboxyarabinitol bisphosphate (CABP). Incorporation of Mn(2+) in the model complex permits investigation of the environment of the activator cation by electron spin resonance (ESR)techniques. Measurements at 9 GHz on the Mn(2+)-containing complex prepared by using dimeric Rhodospirillum rubrum enzyme produce a spectrum which indicates that the cation is bound in an anisotropic environment. Measurements at 9 GHz on the spinach enzyme model complex produce a spectrum in which several of the fine structure transitions are obvious. In contrast, the spectrum produced from Mn(2+) bound to R. rubrum enzyme exhibits an intense powder pattern for the central fine structure transition; the other four fine structure transitions produce powder patterns that are in homogeneously broadened and therefore are not as apparent.Low-temperature measurements at high field (35 GHz) result in substantially simplified spectra. The spectrum of Mn(2+) bound to the R. rubrum enzyme shows less fine structure than the spectrum of Mn(2+) bound in the octameric spinach enzyme complex, where substantial hyperfine splitting is resolved in three of the five fine structure transitions. Measurements at 35 GHz on Mn (2+) bound in the dimeric R. rubrum enzyme complex produce spectra in which only the central fine structure transition produces a prominent signal. However, these samples are characterized by several narrow spectral features which permit investigation of the identity of Mn(2+)ligands by 170 perturbation techniques. Preparation of the R. rubrum RuBP carboxylase model complex in (17)O-enriched water results in a sample which exhibits an obviously broadened 35-GHz Mn(2+) spectrum in comparison to unenriched samples. Removal of H(2)(17)O by gel filtration abolished the spectral

  12. Molecular evolution of urea amidolyase and urea carboxylase in fungi

    OpenAIRE

    Harris Steven D; Nickerson Kenneth W; Strope Pooja K; Moriyama Etsuko N

    2011-01-01

    Abstract Background Urea amidolyase breaks down urea into ammonia and carbon dioxide in a two-step process, while another enzyme, urease, does this in a one step-process. Urea amidolyase has been found only in some fungal species among eukaryotes. It contains two major domains: the amidase and urea carboxylase domains. A shorter form of urea amidolyase is known as urea carboxylase and has no amidase domain. Eukaryotic urea carboxylase has been found only in several fungal species and green al...

  13. Structure of Toxoplasma gondii fructose-1,6-bisphosphate aldolase

    Energy Technology Data Exchange (ETDEWEB)

    Boucher, Lauren E.; Bosch, Jürgen, E-mail: jbosch@jhu.edu [Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205 (United States); Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205 (United States)

    2014-08-29

    The structure of T. gondii fructose-1,6-bisphosphate aldolase, a glycolytic enzyme and structural component of the invasion machinery, was determined to a resolution of 2.0 Å. The apicomplexan parasite Toxoplasma gondii must invade host cells to continue its lifecycle. It invades different cell types using an actomyosin motor that is connected to extracellular adhesins via the bridging protein fructose-1,6-@@bisphosphate aldolase. During invasion, aldolase serves in the role of a structural bridging protein, as opposed to its normal enzymatic role in the glycolysis pathway. Crystal structures of the homologous Plasmodium falciparum fructose-1,6-bisphosphate aldolase have been described previously. Here, T. gondii fructose-1,6-bisphosphate aldolase has been crystallized in space group P22{sub 1}2{sub 1}, with the biologically relevant tetramer in the asymmetric unit, and the structure has been determined via molecular replacement to a resolution of 2.0 Å. An analysis of the quality of the model and of the differences between the four chains in the asymmetric unit and a comparison between the T. gondii and P. falciparum aldolase structures is presented.

  14. Structure of Toxoplasma gondii fructose-1,6-bisphosphate aldolase

    International Nuclear Information System (INIS)

    The structure of T. gondii fructose-1,6-bisphosphate aldolase, a glycolytic enzyme and structural component of the invasion machinery, was determined to a resolution of 2.0 Å. The apicomplexan parasite Toxoplasma gondii must invade host cells to continue its lifecycle. It invades different cell types using an actomyosin motor that is connected to extracellular adhesins via the bridging protein fructose-1,6-@@bisphosphate aldolase. During invasion, aldolase serves in the role of a structural bridging protein, as opposed to its normal enzymatic role in the glycolysis pathway. Crystal structures of the homologous Plasmodium falciparum fructose-1,6-bisphosphate aldolase have been described previously. Here, T. gondii fructose-1,6-bisphosphate aldolase has been crystallized in space group P22121, with the biologically relevant tetramer in the asymmetric unit, and the structure has been determined via molecular replacement to a resolution of 2.0 Å. An analysis of the quality of the model and of the differences between the four chains in the asymmetric unit and a comparison between the T. gondii and P. falciparum aldolase structures is presented

  15. The genes encoding the biotin carboxyl carrier protein and biotin carboxylase subunits of Bacillus subtilis acetyl coenzyme A carboxylase, the first enzyme of fatty acid synthesis.

    OpenAIRE

    P. Marini(GANIL); Li, S J; Gardiol, D; Cronan, J E; De Mendoza, D

    1995-01-01

    The genes encoding two subunits of acetyl coenzyme A carboxylase, biotin carboxyl carrier protein, and biotin carboxylase have been cloned from Bacillus subtilis. DNA sequencing and RNA blot hybridization studies indicated that the B. subtilis accB homolog which encodes biotin carboxyl carrier protein, is part of an operon that includes accC, the gene encoding the biotin carboxylase subunit of acetyl coenzyme A carboxylase.

  16. Interaction Between the Biotin Carboxyl Carrier Domain and the Biotin Carboxylase Domain in Pyruvate Carboxylase from Rhizobium etli†

    OpenAIRE

    Lietzan, Adam D.; Menefee, Ann L.; Zeczycki, Tonya N.; Kumar, Sudhanshu; Attwood, Paul V.; Wallace, John C.; Cleland, W. Wallace; Maurice, Martin St.

    2011-01-01

    Pyruvate carboxylase (PC) catalyzes the ATP-dependent carboxylation of pyruvate to oxaloacetate, an important anaplerotic reaction in mammalian tissues. To effect catalysis, the tethered biotin of PC must gain access to active sites in both the biotin carboxylase domain and the carboxyl transferase domain. Previous studies have demonstrated that a mutation of threonine 882 to alanine in PC from Rhizobium etli renders the carboxyl transferase domain inactive and favors the positioning of bioti...

  17. Structural analysis, plastid localization, and expression of the biotin carboxylase subunit of acetyl-coenzyme A carboxylase from tobacco.

    Science.gov (United States)

    Shorrosh, B S; Roesler, K R; Shintani, D; van de Loo, F J; Ohlrogge, J B

    1995-06-01

    Acetyl-coenzyme A carboxylase (ACCase, EC 6.4.1.2) catalyzes the synthesis of malonyl-coenzyme A, which is utilized in the plastid for de novo fatty acid synthesis and outside the plastid for a variety of reactions, including the synthesis of very long chain fatty acids and flavonoids. Recent evidence for both multifunctional and multisubunit ACCase isozymes in dicot plants has been obtained. We describe here the isolation of a tobacco (Nicotiana tabacum L. cv bright yellow 2 [NT1]) cDNA clone (E3) that encodes a 58.4-kD protein that shares 80% sequence similarity and 65% identity with the Anabaena biotin carboxylase subunit of ACCase. Similar to other biotin carboxylase subunits of acetyl-CoA carboxylase, the E3-encoded protein contains a putative ATP-binding motif but lacks a biotin-binding site (methionine-lysine-methionine or methionine-lysine-leucine). The deduced protein sequence contains a putative transit peptide whose function was confirmed by its ability to direct in vitro chloroplast uptake. The subcellular localization of this biotin carboxylase has also been confirmed to be plastidial by western blot analysis of pea (Pisum sativum), alfalfa (Medicago sativa L.), and castor (Ricinus communis L.) plastid preparations. Northern blot analysis indicates that the plastid biotin carboxylase transcripts are expressed at severalfold higher levels in castor seeds than in leaves. PMID:7610168

  18. Transcriptional activation of phosphoenolpyruvate carboxylase by phosphorus deficiency in tobacco.

    Science.gov (United States)

    Toyota, Kentaro; Koizumi, Nozomu; Sato, Fumihiko

    2003-03-01

    Phosphoenolpyruvate carboxylase (PEPC), which catalyses the carboxylation of phosphoenolpyruvate using HCO(3)(-) to generate oxaloacetic acid, is an important enzyme in the primary metabolism of plants. Although the PEPC genes (ppc) comprise only a small gene family, the function of each gene is not clear, except for roles in C(4) photosynthesis and CAM. Three PEPC genes (Nsppc1-3) from the C(3) plant Nicotiana sylvestris were used to investigate their roles and regulation in a C(3) plant, and their regulation by phosphorus depletion in particular. First, the induction of PEPC by phosphorus depletion was confirmed. Next, Nsppc1 was determined to be mainly responsive to phosphorus deficiency at the transcriptional level. Further studies using transgenic tobacco harbouring a chimeric gene consisting of the 2.0 kb promoter region of Nsppc1 and the beta-glucuronidase (GUS) reporter showed that PEPC is transcriptionally induced. It was also found that sucrose had a synergistic effect on the induction of PEPC by phosphorus deficiency. A series of transgenic tobacco containing 5'-deletion mutants of Nsppc1 promoter::GUS fusion revealed that the -539 to -442 bp Nsppc1 promoter region, relative to the translation start site, was necessary for the response to phosphorus deficiency. Gain-of-function analysis using a construct containing three tandem repeats of the -539 to -442 bp region confirmed that this region was sufficient to induce the phosphorus-deficiency response in tobacco. PMID:12598567

  19. The source and characteristics of chemiluminescence associated with the oxygenase reaction catalyzed by Mn(2+)-ribulosebisphosphate carboxylase.

    Science.gov (United States)

    Lilley, R M; Riesen, H; Andrews, T J

    1993-07-01

    We confirm the observation of Mogel and McFadden (Mogel, S.N., and McFadden, B. A. (1990) Biochemistry 29, 8333-8337) that ribulosebisphosphate carboxylase/oxygenase (rubisco) exhibits chemiluminescence while catalyzing its oxygenase reaction in the presence of Mn2+. However, our results with the spinach and Rhodospirillum rubrum enzymes differ markedly in the following respects. 1) Chemiluminescence intensity was directly proportional to enzyme concentration and behaved as if representing the rate of oxygenase catalysis. 2) The wavelength spectrum peaked at about 770 nm and extended beyond 810 nm. This seems inconsistent with chemiluminescence generated by simultaneous decay of pairs of singlet O2 molecules. It is consistent with manganese(II) luminescence and we discuss its possible sources. The time course of chemiluminescence (resolution, 0.25 s) was distinctively different for spinach and R. rubrum enzymes during the initial 5 s of catalysis, with the bacterial enzyme exhibiting a pronounced initial "burst." Chemiluminescence by the spinach enzyme responded to substrate concentrations in a manner consistent with known oxygenase properties, exhibiting Michaelis-Menten kinetics with ribulose-1,5-bisphosphate (Km 400 nM). Chemiluminescence required carbamylated enzyme with Mn2+ bound at the active site (activation energy, -57.1 KJ.mol-1). As an indicator of oxygenase activity, chemiluminescence represents an improvement over oxygen electrode measurements in response time and sensitivity by factors of at least 100. PMID:8314755

  20. Regulation of phosphoenolpyruvate carboxylase in PVYNTN-infected tobacco plants

    Czech Academy of Sciences Publication Activity Database

    Müller, Karel; Doubnerová, V.; Synková, Helena; Čeřovská, Noemi; Ryšlavá, H.

    2009-01-01

    Roč. 390, č. 3 (2009), s. 245-251. ISSN 1431-6730 R&D Projects: GA ČR GA206/03/0310; GA MŠk 1M0505 Institutional research plan: CEZ:AV0Z50380511 Keywords : biotic stress * Nicotiana tabacum * phosphoenolpyruvate carboxylase (PEPC) Subject RIV: ED - Physiology Impact factor: 2.732, year: 2009

  1. Structure of fructose bisphosphate aldolase from Encephalitozoon cuniculi

    International Nuclear Information System (INIS)

    The eukaryotic parasite E. cuniculi expresses a fructose bisphosphate aldolase that crystallizes readily in the presence of the partial substrate analog phosphate. This aldolase–phosphate structure and that of the sugar-bound Schiff base are reported. E. cuniculi aldolase displays a dimeric structure rather than the expected tetrameric quaternary structure. Fructose bisphosphate aldolose (FBPA) enzymes have been found in a broad range of eukaryotic and prokaryotic organisms. FBPA catalyses the cleavage of fructose 1,6-bisphosphate into glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. The SSGCID has reported several FBPA structures from pathogenic sources. Bioinformatic analysis of the genome of the eukaryotic microsporidian parasite Encephalitozoon cuniculi revealed an FBPA homolog. The structures of this enzyme in the presence of the native substrate FBP and also with the partial substrate analog phosphate are reported. The purified enzyme crystallized in 90 mM Bis-Tris propane pH 6.5, 18% PEG 3350, 18 mM NaKHPO4, 10 mM urea for the phosphate-bound form and 100 mM Bis-Tris propane pH 6.5, 20% PEG 3350, 20 mM fructose 1,6-bisphosphate for the FBP-bound form. In both cases protein was present at 25 mg ml−1 and the sitting-drop vapour-diffusion method was used. For the FBP-bound form, a data set to 2.37 Å resolution was collected from a single crystal at 100 K. The crystal belonged to the orthorhombic space group C2221, with unit-cell parameters a = 121.46, b = 135.82, c = 61.54 Å. The structure was refined to a final free R factor of 20.8%. For the phosphate-bound form, a data set was collected to 2.00 Å resolution. The space group was also C2221 and the unit-cell parameters were a = 121.96, b = 137.61, c = 62.23 Å. The structure shares the typical barrel tertiary structure reported for previous FBPA structures and exhibits the same Schiff base in the active site. The quaternary structure is dimeric. This work provides a direct experimental result

  2. UniProt search blastx result: AK288054 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288054 J075152E14 P29684|RBS_HEVBR Ribulose bisphosphate carboxylase small chain, chloroplast ... (RuBisCO small subunit) - Hevea brasiliensis (Para rubber ... tree) 7.00E-62 ...

  3. UniProt search blastx result: AK287599 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287599 J065050G24 P29684|RBS_HEVBR Ribulose bisphosphate carboxylase small chain, chloroplast ... (RuBisCO small subunit) - Hevea brasiliensis (Para rubber ... tree) 1.00E-61 ...

  4. UniProt search blastx result: AK288517 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288517 J090043B03 P29684|RBS_HEVBR Ribulose bisphosphate carboxylase small chain, chloroplast ... (RuBisCO small subunit) - Hevea brasiliensis (Para rubber ... tree) 1.00E-55 ...

  5. Phosphatidylinositol(4,5)bisphosphate and phosphatidylinositol(4)phosphate in plant tissues. [Pisum sativum

    Energy Technology Data Exchange (ETDEWEB)

    Irvine, R.F.; Letcher, A.J.; Lander, D.J. (Institute of Animal Physiology and Genetics Research, Cambridge (England)); Dawson, A.P. (Univ. of East Anglia, Norwich (England)); Musgrave, A. (Univ. of Amsterdam (Netherlands)); Drobak, B.K. (John Innes Institute, Norwich (England))

    1989-03-01

    Pea (Pisum sativum) leaf discs or swimming suspensions of Chlamydomonas eugametos were radiolabeled with ({sup 3}H)myo-inositol or ({sup 32}P)Pi and the lipids were extracted, deacylated, and their glycerol moieties removed. The resulting inositol trisphosphate and bisphosphate fractions were examined by periodate degradation, reduction and dephosphorylation, or by incubation with human red cell membranes. Their likely structures were identified as D-myo-inositol(1,4,5)trisphosphate and D-myo-inositol(1,4,)-bisphosphate. It is concluded that plants contain phosphatidylinositol(4)phosphate and phosphatidylinositol(4,5)bisphosphate; no other polyphosphoinositides were detected.

  6. Mechanisms of Inhibition of Rhizobium etli Pyruvate Carboxylase by l-Aspartate

    OpenAIRE

    Sirithanakorn, Chaiyos; Adina-Zada, Abdussalam; Wallace, John C.; Jitrapakdee, Sarawut; Attwood, Paul V.

    2014-01-01

    l-Aspartate is a regulatory feedback inhibitor of the biotin-dependent enzyme pyruvate carboxylase in response to increased levels of tricarboxylic acid cycle intermediates. Detailed studies of l-aspartate inhibition of pyruvate carboxylase have been mainly confined to eukaryotic microbial enzymes, and aspects of its mode of action remain unclear. Here we examine its inhibition of the bacterial enzyme Rhizobium etli pyruvate carboxylase. Kinetic studies demonstrated that l-aspartate binds to ...

  7. Long-term effect of elevated CO2 on spatial differentiation of ribulose-1,5-bisphosphate carboxylase/oxygenase activity in Norway spruce canopy

    Czech Academy of Sciences Publication Activity Database

    Hrstka, M.; Urban, Otmar; Marek, Michal V.

    2005-01-01

    Roč. 43, č. 2 (2005), s. 211-216. ISSN 0300-3604 R&D Projects: GA MŠk(CZ) LN00A141; GA MŽP(CZ) SM/640/18/03; GA MŠk(CZ) OC E21.001 Institutional research plan: CEZ:AV0Z6087904 Keywords : down-regulation * photosynthesis * sun and shade needles * vertical profile Subject RIV: ED - Physiology Impact factor: 0.810, year: 2005

  8. Impact of ozone on the activity and quantity of ribulose bisphosphate carboxylase/oxygenase in potato foliage and its relation to premature senescence

    Energy Technology Data Exchange (ETDEWEB)

    Dann, M.S.

    1988-01-01

    Plants, 26 days old, were exposed to O{sub 3} for five days in a controlled environment chamber. Initial and total activities and quantity of enzyme declined in both O{sub 3}-treated and control plants. Ozone accelerated the decline and produced a significantly greater decrease in activity and quantity by the fifth day of O{sub 3} exposure. Percent activation of the enzyme did not change. Enzyme activity and quantity of O{sub 3}-treated plants remained below control levels throughout a seven-day post-exposure observation period. Ozone may accelerate the decline in rubisco by increasing enzyme acceptability to proteases. To test this hypothesis, rubisco was purified from Norland potato foliage and the enzyme exposed to O{sub 3} or oxygen (O{sub 2}) in vitro. Following oxidant treatment, the extract was treated with exogenous protease. Oxidant treatment reduced the quantity of rubisco an average of 15%. Addition of protease further reduced the quantity by another 30% after O{sub 3} but no O{sub 2} treatment.

  9. Changes in mRNA levels and polypeptide subunits of ribulose 1,5-bisphosphate carboxylase in response to supplementary ultraviolet-B radiation

    International Nuclear Information System (INIS)

    Pea plants (Pisum sativum L., cv. Greenfeast) were grown for 17d (150 μmol photons m−2 s−1; 12h light/12 h dark) and then exposed to moderate levels of supplementary ultraviolet-B radiation (UV-B: 280–320 nm) during the light cycle. The total soluble leaf protein, maximum Rubisco activity, polypeptide and mRNA transcript levels for Rubisco subunits were then determined in the mature third leaf pair from the base of the plants. Total soluble protein per unit leaf area showed little change after 1 d but declined by 33% during 3d of UV-B exposure. However, there was no change on a unit chlorophyll basis. Total RNA per unit area declined by 15% and 37% after 1 or 3d of UV-B treatment, respectively. Maximum Rubisco activity declined by 38% after 1 d and 71% after 3d of UV-B exposure. Rubisco polypeptide subunits showed some decrease (∼16%) after 1d exposure, but declined by 56% over 3d. The decrease in Rubisco is probably the major reason for the reduction in soluble protein. In contrast to the relatively slow decline in total soluble protein and Rubisco, the level of the mRNA transcripts for both rbc L and rbc S showed a dramatic decrease within hours of UV-B exposure. The mRNA transcripts for rbc S were reduced to >20% of control values after 4h of UV-B exposure, while the rbc L transcripts were reduced by 60% after 8h. Further exposure to UV-B reduced the mRNA transcripts to either trace or undetectable levels. The decrease in rbc S mRNA levels with the UV-B exposure can be partially ameliorated by higher photosynthetically active irradiance during the period of UV-B exposure. Plants that were exposed to supplementary UV-B radiation for short periods (4h or 8h) and returned to control conditions, showed no recovery after 24h. However, after a further 2d, the rbc L and rbc S mRNA transcripts had recovered to ca. 60% of the control values, showing that the effect upon the mRNA transcripts is a reversible response. (author)

  10. Control of liver 6-phosphofructokinase by fructose 2,6-bisphosphate and other effectors.

    OpenAIRE

    Van Schaftingen, Emile; Jett, M F; Hue, Louis; Hers, H G

    1981-01-01

    Rat liver 6-phosphofructokinase (ATP-D-fructose-6-phosphate 1-phosphotransferase, EC 2.7.1.11) was partially purified free of interfering enzymes by a rapid procedure. Fructose 2,6-bisphosphate, at micromolar concentrations, greatly stimulated the enzyme by increasing its affinity for fructose 6-phosphate and relieving the inhibition by ATP. Its action was synergistic with that of AMP. As a stimulator of liver phosphofructokinase, fructose 2,6-bisphosphate was approximately 1000- and 2500-fol...

  11. Regulation of fructose 2,6-bisphosphate concentration in white adipose tissue.

    OpenAIRE

    Rider, Mark; Hue, Louis

    1985-01-01

    Injection of insulin to fed rats diminished the concentration of fructose 2,6-bisphosphate in white adipose tissue. Incubation of epididymal fat-pads or adipocytes with insulin stimulated lactate release and sugar detritiation and also decreased fructose 2,6-bisphosphate concentration. Such a decrease was, however, not observed in fat-pads from starved or alloxan-diabetic rats. Incubation of adipocytes from fed rats with various concentrations of glucose or fructose led to a dose-dependent ri...

  12. UniProt search blastx result: AK288054 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288054 J075152E14 P16134|RBS1_ACEAT Ribulose bisphosphate carboxylase small chain 1, chloropla ... ll subunit 1) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 1.00E-31 ...

  13. UniProt search blastx result: AK288054 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288054 J075152E14 P16137|RBS4_ACEAT Ribulose bisphosphate carboxylase small chain 4, chloropla ... ll subunit 4) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 3.00E-31 ...

  14. UniProt search blastx result: AK287599 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287599 J065050G24 P16138|RBS5_ACEAT Ribulose bisphosphate carboxylase small chain 5, chloropla ... ll subunit 5) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 6.00E-32 ...

  15. UniProt search blastx result: AK288517 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288517 J090043B03 P16137|RBS4_ACEAT Ribulose bisphosphate carboxylase small chain 4, chloropla ... ll subunit 4) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 9.00E-31 ...

  16. UniProt search blastx result: AK288054 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288054 J075152E14 P16138|RBS5_ACEAT Ribulose bisphosphate carboxylase small chain 5, chloropla ... ll subunit 5) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 4.00E-32 ...

  17. UniProt search blastx result: AK288517 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288517 J090043B03 P16136|RBS3_ACEAT Ribulose bisphosphate carboxylase small chain 3, chloropla ... ll subunit 3) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 1.00E-31 ...

  18. UniProt search blastx result: AK288517 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288517 J090043B03 P16138|RBS5_ACEAT Ribulose bisphosphate carboxylase small chain 5, chloropla ... ll subunit 5) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 7.00E-31 ...

  19. UniProt search blastx result: AK288517 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288517 J090043B03 P16134|RBS1_ACEAT Ribulose bisphosphate carboxylase small chain 1, chloropla ... ll subunit 1) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 2.00E-31 ...

  20. UniProt search blastx result: AK288517 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288517 J090043B03 P16135|RBS2_ACEAT Ribulose bisphosphate carboxylase small chain 2, chloropla ... 2) (Fragment) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 2.00E-31 ...

  1. UniProt search blastx result: AK287599 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287599 J065050G24 P16137|RBS4_ACEAT Ribulose bisphosphate carboxylase small chain 4, chloropla ... ll subunit 4) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 5.00E-31 ...

  2. UniProt search blastx result: AK288054 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288054 J075152E14 P16136|RBS3_ACEAT Ribulose bisphosphate carboxylase small chain 3, chloropla ... ll subunit 3) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 8.00E-32 ...

  3. UniProt search blastx result: AK287599 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287599 J065050G24 P16134|RBS1_ACEAT Ribulose bisphosphate carboxylase small chain 1, chloropla ... ll subunit 1) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 2.00E-31 ...

  4. UniProt search blastx result: AK287599 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287599 J065050G24 P16136|RBS3_ACEAT Ribulose bisphosphate carboxylase small chain 3, chloropla ... ll subunit 3) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 1.00E-31 ...

  5. UniProt search blastx result: AK288054 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288054 J075152E14 P16135|RBS2_ACEAT Ribulose bisphosphate carboxylase small chain 2, chloropla ... 2) (Fragment) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 1.00E-31 ...

  6. UniProt search blastx result: AK287599 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287599 J065050G24 P16135|RBS2_ACEAT Ribulose bisphosphate carboxylase small chain 2, chloropla ... 2) (Fragment) - Acetabularia acetabulum (Mermaid's wine ... glass) (Acetabularia mediterranea) 2.00E-31 ...

  7. The dynamic organization of fungal acetyl-CoA carboxylase

    Science.gov (United States)

    Hunkeler, Moritz; Stuttfeld, Edward; Hagmann, Anna; Imseng, Stefan; Maier, Timm

    2016-04-01

    Acetyl-CoA carboxylases (ACCs) catalyse the committed step in fatty-acid biosynthesis: the ATP-dependent carboxylation of acetyl-CoA to malonyl-CoA. They are important regulatory hubs for metabolic control and relevant drug targets for the treatment of the metabolic syndrome and cancer. Eukaryotic ACCs are single-chain multienzymes characterized by a large, non-catalytic central domain (CD), whose role in ACC regulation remains poorly characterized. Here we report the crystal structure of the yeast ACC CD, revealing a unique four-domain organization. A regulatory loop, which is phosphorylated at the key functional phosphorylation site of fungal ACC, wedges into a crevice between two domains of CD. Combining the yeast CD structure with intermediate and low-resolution data of larger fragments up to intact ACCs provides a comprehensive characterization of the dynamic fungal ACC architecture. In contrast to related carboxylases, large-scale conformational changes are required for substrate turnover, and are mediated by the CD under phosphorylation control.

  8. Studies on biotin dependent carboxylases and the properties of carboxybiotin

    International Nuclear Information System (INIS)

    Biotin dependent carboxyl-transfer reactions have been studied using biotin and pyruvate carboxylases. The pH profile for the Mg2+ and MgATP dependent carboxylation of biotin by bicarbonate shows that an enzymic base with a pK of 6.5 must be unprotonated for catalysis to occur. The pH profiles for the carboxyl-transfer reaction of pyruvate carboxylase have been obtained by studying the decarboxylation of oxalacetate stimulated by the presence of oxamate. Similarly, 13C and 2H isotope effects have been measured for the decarboxylation of oxalacetate by both enzymic and nonenzymic means. From these studies the authors can conclude that carboxyl-transfer between biotin and oxalacetate is at least partially rate-limiting and is not concerted with proton-transfer. The lack of any apparent enzymic acid-base catalyst (the V/K profile for oxalacetate is pH independent) suggests that proton transfer may occur directly between biotin and the carbanion formed when oxalacetate is decarboxylated. The pH profile for the nonenzymatic decarboxylation of carboxybiotin shows a plateau below pH 4 (k = 0.012 min-1 at 20C), and a lower plateau above pH 8 (k = 0.005 min-1 at 250C). A proton inventory at low pH is linear, while at high pH it is curved. These data suggest that two different mechanisms operate at high and low pH

  9. Fructose 1,6-bisphosphate aldolase activity is essential for synthesis of alginate from glucose by Pseudomonas aeruginosa.

    OpenAIRE

    Banerjee, P C; Vanags, R I; Chakrabarty, A M; Maitra, P. K.

    1985-01-01

    We have isolated a mutant of Pseudomonas aeruginosa deficient in fructose 1,6-bisphosphate aldolase activity. This mutant, similar to the mutants deficient in any of the Entner-Doudoroff pathway enzymes, does not allow appreciable alginate formation from glucose and gluconate, but allows alginate synthesis from mannitol and fructose. This suggests that glucose and gluconate must be converted to fructose 1,6-bisphosphate via the Entner-Doudoroff pathway enzymes and fructose 1,6-bisphosphate al...

  10. Influence of nitrogen enrichment on size-fractionated in vitro carboxylase activities of phytoplankton from Thau Lagoon (Coastal Mediterranean Lagoon, France)

    OpenAIRE

    Fouilland, Eric; Descolas Gros, Chantal; Collos, Yves; Vaquer A, André; Souchu, Philippe; Gasc, Anne; Bibent, Bertrand; Pons, Virginie

    2002-01-01

    The influence of dissolved inorganic and organic nitrogen (DIN and DON) enrichments on pools of enzymes responsible for CO2 fixation by the Calvin-Benson (Rubisco) and beta-carboxylation pathways (beta-carboxylases) were studied in a natural plankton assemblage. The plankton community from a coastal Mediterranean lagoon were incubated in situ for 24 h with initially ammonium, nitrate and DON (taurine) enrichments and compared to a control without any enrichment. An increase of small picophyto...

  11. Increased expression of fatty acid synthase and acetyl-CoA carboxylase in the prefrontal cortex and cerebellum in the valproic acid model of autism

    Science.gov (United States)

    Chen, Jianling; Wu, Wei; Fu, Yingmei; Yu, Shunying; Cui, Donghong; Zhao, Min; Du, Yasong; Li, Jijun; Li, Xiaohong

    2016-01-01

    The primary aim of the present study was to investigate alterations in enzymes associated with fatty acid synthesis, namely fatty acid synthase (FASN) and acetyl-CoA carboxylase (ACC), in the prefrontal cortex and cerebellum of the valproic acid (VPA)-induced animal model of autism. In this model, pregnant rats were given a single intraperitoneal injection of VPA, and prefrontal cortex and cerebellum samples from their pups were analyzed. The results of western blotting and reverse transcription-quantitative polymerase chain reaction analyses demonstrated that the protein and mRNA expression levels of FASN, ACC and phospho-ACC (pACC) were increased in the prefrontal cortex and cerebellum of the VPA model of autism. Furthermore, in the prefrontal cortex and cerebellum of the VPA model of autism, AMPK expression is increased, whereas PI3K and Akt expression are unchanged. This suggests that disorder of the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/Akt/FASN and/or adenosine 5′-monophosphate-activated protein kinase (AMPK)/ACC pathway may be involved in the pathogenesis of autism. It is hypothesized that fatty acid synthesis participates in autism through PI3K/Akt/FASN and AMPK/ACC pathways. PMID:27602061

  12. Biotin Responsive Multiple Carboxylase Deficiency Presenting as Diabetic Ketoacidosis.

    Directory of Open Access Journals (Sweden)

    Jia-Woei Hou

    2004-02-01

    Full Text Available Multiple carboxylase deficiency (MCD is a rare inherited metabolic disease of biotindependency due to deficiency of holocarboxylase synthetase (HCS or biotinidase deficiency.A 30-month-old female patient who presented with the initial features of diabeticketoacidosis (severe metabolic acidosis, ketosis, and hyperglycemia, lactic acidemia, moderatehyperammonemia, and generalized organic aciduria is described. Associated symptomsand signs included erythematous skin rashes, alopecia and developmental delay. The patientresponded dramatically to treatment with biotin (10 mg/day showing normalization of clinicalsymptoms and most biochemical abnormalities. Based on the urine organic profile bygas chromatography/ mass spectrometry (GC/MS, the diagnosis of MCD was made. A plasmatandem mass study confirmed this diagnosis. The biotinase activity in serum was normal,indicating that this was a rare case of late-onset HCS deficiency.

  13. Decreased renal vitamin K-dependent γ-glutamyl carboxylase activity in calcium oxalate calculi patients

    Institute of Scientific and Technical Information of China (English)

    陈俊汇; 刘继红; 章咏裳; 叶章群; 王少刚

    2003-01-01

    Objective To study the activity of vitamin K-dependent γ-glutamyl carboxylase in patients with calcium oxalate (CaOx) urolithiasis compared with healthy individuals and to assess its relationship to the renal calcium oxalate urolithiasis. Methods Renal parenchymas were harvested from urolithic patients and renal tumor patients undergoing nephrectomy. The renal carboxylase activity was evaluated as the radioactivity of [14C] labeled sodium bicarbonate in carboxylic reactions in vitro using β-liquid scintillation counting. Results Significantly reduced activity of renal vitamin K-dependent γ-glutamyl carboxylase was observed in the urolithic group as compared with normal controls (P<0.01). Conclusion It suggests that the reduced carboxylase activity observed in the urolithic patients may play an important role in the course of renal calcium oxalate urolithiasis.

  14. Effect of Pyruvate Carboxylase Overexpression on the Physiology of Corynebacterium glutamicum

    OpenAIRE

    Koffas, Mattheos A. G.; Jung, Gyoo Yeol; Aon, Juan C.; Stephanopoulos, Gregory

    2002-01-01

    Pyruvate carboxylase was recently sequenced in Corynebacterium glutamicum and shown to play an important role of anaplerosis in the central carbon metabolism and amino acid synthesis of these bacteria. In this study we investigate the effect of the overexpression of the gene for pyruvate carboxylase (pyc) on the physiology of C. glutamicum ATCC 21253 and ATCC 21799 grown on defined media with two different carbon sources, glucose and lactate. In general, the physiological effects of pyc overe...

  15. Modification of Rhizopus lactate dehydrogenase for improved resistance to fructose 1,6-bisphosphate

    Science.gov (United States)

    Rhizopus oryzae is frequently used for fermentative production of lactic acid. We determined that one of the key enzymes, lactate dehydrogenase (LDH), involved in synthesis of lactic acid by R. oryzae was significantly inhibited by fructose 1,6-bisphosphate (FBP) at physiological concentrations. Thi...

  16. Pyruvate carboxylase deficiency: An underestimated cause of lactic acidosis

    Directory of Open Access Journals (Sweden)

    F. Habarou

    2015-03-01

    Full Text Available Pyruvate carboxylase (PC is a biotin-containing mitochondrial enzyme that catalyzes the conversion of pyruvate to oxaloacetate, thereby being involved in gluconeogenesis and in energy production through replenishment of the tricarboxylic acid (TCA cycle with oxaloacetate. PC deficiency is a very rare metabolic disorder. We report on a new patient affected by the moderate form (the American type A. Diagnosis was nearly fortuitous, resulting from the revision of an initial diagnosis of mitochondrial complex IV (C IV defect. The patient presented with severe lactic acidosis and pronounced ketonuria, associated with lethargy at age 23 months. Intellectual disability was noted at this time. Amino acids in plasma and organic acids in urine did not show patterns of interest for the diagnostic work-up. In skin fibroblasts PC showed no detectable activity whereas biotinidase activity was normal. We had previously reported another patient with the severe form of PC deficiency and we show that she also had secondary C IV deficiency in fibroblasts. Different anaplerotic treatments in vivo and in vitro were tested using fibroblasts of both patients with 2 different types of PC deficiency, type A (patient 1 and type B (patient 2. Neither clinical nor biological effects in vivo and in vitro were observed using citrate, aspartate, oxoglutarate and bezafibrate. In conclusion, this case report suggests that the moderate form of PC deficiency may be underdiagnosed and illustrates the challenges raised by energetic disorders in terms of diagnostic work-up and therapeutical strategy even in a moderate form.

  17. Genes encoding biotin carboxylase subunit of acetyl-CoA carboxylase from Brassica napus and parental species: cloning, expression patterns, and evolution

    Science.gov (United States)

    Comparative genomics is a useful tool to investigate gene and genome evolution. Biotin carboxylase (BC), an important subunit of heteromeric ACCase that is a rate-limiting enzyme in fatty acid biosynthesis in dicots, catalyzes ATP, biotin-carboxyl-carrier protein and CO2 to form carboxybiotin-carbo...

  18. Characterization of three protein components required for functional reconstitution of the epoxide carboxylase multienzyme complex from Xanthobacter strain Py2.

    OpenAIRE

    Allen, J. R.; Ensign, S A

    1997-01-01

    Epoxide carboxylase from Xanthobacter strain Py2 catalyzes the reductant- and NAD+-dependent carboxylation of aliphatic epoxides to beta-keto acids. Epoxide carboxylase from Xanthobacter strain Py2 has been resolved from cell extracts by anion-exchange chromatography into three protein components, designated I, II, and III, that are obligately required for functional reconstitution of epoxide carboxylase activity. Component II has been purified to homogeneity on the basis of its ability to co...

  19. Fructose 2,6-bisphosphate (Fru-2,6-P2) binds at the active site of rabbit liver fructose-1,6-bisphosphate

    International Nuclear Information System (INIS)

    Fru-2,6-P2 and AMP are potent inhibitors of fructose-1,6-bisphosphatase (FBPase). AMP inhibits by binding to an allosteric site. While Fru-2,6-P2 and fructose 1,6-bisphosphate (Fru-1,6-P2) binding is mutually exclusive, a controversy exists as to whether Fru-2,6-P2 inhibits by binding to the active site or a regulatory site. To address this question, the aromatic region of the 1H NMR spectrum of FBPase was examined in the absence and presence of Fru-1,6-P2, Fru-2,6-P2, and AMP. All of the spectral perturbations produced by Fru-1,6-P2 binding were also seen with Fru-2,6-P2: (1) the C2-H of His 1 (pKa=7.1) shifted downfield 0.03 ppm (pH 5.3); (2) the C2-H of His 2 shifted downfield 0.03 ppm (pH 5.3) and broadened; (3) several resonances in the phe/tyr region of the spectrum decreased in intensity. In addition, both fructose bisphosphates produced a change in the exchange rate of AMP from past/intermediate in the binary enzyme AMP complex to slow exchange in the ternary complex. AMP produced similar changes in the phe/tyr region of the spectrum. Unlike the fructose bisphosphates, AMP binding produced: (1) broadening of the C2-H of His 1; (2) no effect on His 2 and; (3) an enhancement of the intensity of a resonance at 7.25 ppm (pH 5.3). The results are consistent with an active site binding mode of inhibition for Fru-2,6-P2. AMP binding to a regulatory site was apparent from the different results obtained with this ligand

  20. Increased expression of pyruvate carboxylase and biotin protein ligase increases lysine production in a biotin prototrophic Corynebacterium glutamicum strain

    DEFF Research Database (Denmark)

    Wang, Zhihao; Moslehi-Jenabian, Soloomeh; Solem, Christian;

    2015-01-01

    -CoA (or pimeloyl-Acyl Carrier Protein [ACP]) formation. Pyruvate carboxylase (pycA), a biotin-dependent enzyme needed for lysine biosynthesis and biotin ligase (birA), which is responsible for attaching biotin to pyruvate carboxylase, were overexpressed by replacing the native promoters with the strong...

  1. Carbon-13 and deuterium isotope effects on the catalytic reactions of biotin carboxylase

    International Nuclear Information System (INIS)

    13C and 2H kinetic isotope effects have been used to investigate the mechanism of enzymic biotin carboxylation. /sup D/(V/K) is 0.50 in 80% D2O at pD 8.0 for the forward reaction and 0.57 at pD 8.5 for the phosphorylation of ADP by carbamoyl phosphate. These values approach the theoretical maximum limit for a reaction in which a proton is transferred from a sulfhydryl to a nitrogen or oxygen base. Therefore, it appears that this portion of the reaction is at or near equilibrium. 13(V/K) at pH 8 is 1.007; the small magnitude of this number suggests that the reaction is almost fully committed by the time the carbon-sensitive steps are reached. There does not appear to be a reverse commitment to the reaction under the conditions in which 13(V/K) was determined. A large forward commitment is consistent with the failure to observe positional isotope exchange from the βγ-bridge position to the β-nonbridge position in [18O4]ATP or washout of 18O from the γ-nonbridge positions. Transfer of 18O from bicarbonate to inorganic phosphate in the forward reaction was clearly observed, however. These observations suggest that biotin carboxylase exists in two distinct forms which differ in the protonation states of the two active-site bases, one of which is a sulfhydryl. Only when the sulfhydryl is ionized and the second base protonated can catalysis take place. Carboxylation of biotin is postulated to occur via a pathway in which carboxyphosphate is formed by nucleophilic attack of bicarbonate on ATP. Decarboxylation of carboxyphosphate in the active site generates CO2, which serves to carboxylate the isourea tautomer of biotin that is generated by the removal of the proton on N1' by the ionized sulfhydryl

  2. Characterization of phosphoenolpyruvate carboxylase from mature maize seeds: Properties of phosphorylated and dephosphorylated forms

    Czech Academy of Sciences Publication Activity Database

    Černý, M.; Doubnerová, V.; Müller, Karel; Ryšlavá, H.

    2010-01-01

    Roč. 92, č. 10 (2010), s. 1362-1370. ISSN 0300-9084 R&D Projects: GA MŠk 1M0505 Institutional research plan: CEZ:AV0Z50380511 Keywords : Phosphoenolpyruvate carboxylase * Phosphorylation * Seed Subject RIV: EI - Biotechnology ; Bionics Impact factor: 3.787, year: 2010

  3. AKAP150 mediates TRPV1 sensitivity to phosphatidylinositol-4, 5-bisphosphate

    OpenAIRE

    Jeske, Nathaniel A.; Por, Elaine D.; Belugin, Sergei; Chaudhury, Sraboni; Berg, Kelly A; Akopian, Armen N.; Henry, Michael A.; Gomez, Ruben

    2011-01-01

    A-kinase anchoring protein 150 (AKAP150) is a scaffolding protein that controls protein kinase A- and C-mediated phosphorylation of the transient receptor potential family V type 1 (TRPV1), dictating receptor response to nociceptive stimuli. The phospholipid phosphatidylinositol-4,5-bisphosphate (PIP2) anchors AKAP150 to the plasma membrane in naïve conditions, and also affects TRPV1 activity. In the present study, we sought to determine whether the effects of PIP2 on TRPV1 are mediated throu...

  4. Diffusion Coefficient of Fluorescent Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane of Cells

    OpenAIRE

    Golebiewska, Urszula; Nyako, Marian; Woturski, William; Zaitseva, Irina; McLaughlin, Stuart

    2008-01-01

    Phosphatidylinositol 4,5-bisphosphate (PIP2) controls a surprisingly large number of processes in cells. Thus, many investigators have suggested that there might be different pools of PIP2 on the inner leaflet of the plasma membrane. If a significant fraction of PIP2 is bound electrostatically to unstructured clusters of basic residues on membrane proteins, the PIP2 diffusion constant, D, should be reduced. We microinjected micelles of Bodipy TMR-PIP2 into cells, and we measured D on the inne...

  5. Synthesis and Biological Activity of Phospholipase C-Resistant Analogues of Phosphatidylinositol 4, 5-bisphosphate

    OpenAIRE

    Zhang, Honglu; Xu, Yong; Zhang, Zheng; Liman, Emily R.; Prestwich, Glenn D.

    2006-01-01

    The membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) is an important regulator in cell physiology. Hydrolysis of PtdIns(4,5)P2 by phospholipase C (PLC) releases two second messengers, Ins(1,4,5)P3 and diacylglycerol. To dissect the effects of PtdIns(4,5)P2 from those resulting from PLC-generated signals, a metabolically-stabilized analogue of PtdIns(4,5)P2 was required. Two analogues were designed in which the scissile O-P bond was replaced with a C-P bond that coul...

  6. Rational Design Synthesis and Evaluation of New Selective Inhibitors of Microbial Class II (Zinc Dependent) Fructose Bis-phosphate Aldolases

    Energy Technology Data Exchange (ETDEWEB)

    R Daher; M Coincon; M Fonvielle; P Gest; M Guerin; M Jackson; J Sygusch; M Therisod

    2011-12-31

    We report the synthesis and biochemical evaluation of several selective inhibitors of class II (zinc dependent) fructose bis-phosphate aldolases (Fba). The products were designed as transition-state analogues of the catalyzed reaction, structurally related to the substrate fructose bis-phosphate (or sedoheptulose bis-phosphate) and based on an N-substituted hydroxamic acid, as a chelator of the zinc ion present in active site. The compounds synthesized were tested on class II Fbas from various pathogenic microorganisms and, by comparison, on a mammalian class I Fba. The best inhibitor shows Ki against class II Fbas from various pathogens in the nM range, with very high selectivity (up to 105). Structural analyses of inhibitors in complex with aldolases rationalize and corroborate the enzymatic kinetics results. These inhibitors represent lead compounds for the preparation of new synthetic antibiotics, notably for tuberculosis prophylaxis.

  7. Structure of a Class I Tagatose-1,6-bisphosphate Aldolase - Investigation into an Apparent Loss of Stereospecificity

    Energy Technology Data Exchange (ETDEWEB)

    LowKam, C.; Liotard, B; Sygusch, J

    2010-01-01

    Tagatose-1,6-bisphosphate aldolase from Streptococcus pyogenes is a class I aldolase that exhibits a remarkable lack of chiral discrimination with respect to the configuration of hydroxyl groups at both C3 and C4 positions. The enzyme catalyzes the reversible cleavage of four diastereoisomers (fructose 1,6-bisphosphate (FBP), psicose 1,6-bisphosphate, sorbose 1,6-bisphosphate, and tagatose 1,6-bisphosphate) to dihydroxyacetone phosphate (DHAP) and d-glyceraldehyde 3-phosphate with high catalytic efficiency. To investigate its enzymatic mechanism, high resolution crystal structures were determined of both native enzyme and native enzyme in complex with dihydroxyacetone-P. The electron density map revealed a ({alpha}/{beta}){sub 8} fold in each dimeric subunit. Flash-cooled crystals of native enzyme soaked with dihydroxyacetone phosphate trapped a covalent intermediate with carbanionic character at Lys{sup 205}, different from the enamine mesomer bound in stereospecific class I FBP aldolase. Structural analysis indicates extensive active site conservation with respect to class I FBP aldolases, including conserved conformational responses to DHAP binding and conserved stereospecific proton transfer at the DHAP C3 carbon mediated by a proximal water molecule. Exchange reactions with tritiated water and tritium-labeled DHAP at C3 hydrogen were carried out in both solution and crystalline state to assess stereochemical control at C3. The kinetic studies show labeling at both pro-R and pro-S C3 positions of DHAP yet detritiation only at the C3 pro-S-labeled position. Detritiation of the C3 pro-R label was not detected and is consistent with preferential cis-trans isomerism about the C2-C3 bond in the carbanion as the mechanism responsible for C3 epimerization in tagatose-1,6-bisphosphate aldolase.

  8. Reaction of phosphoenolpyruvate carboxylase with (Z)-3-bromophosphoenolpyruvate and (Z)-3-fluorophosphoenolpyruvate

    International Nuclear Information System (INIS)

    (Z)-3-Bromophosphoenolpyruvate inactivates phosphoenolpyruvate carboxylase from maize in the presence of HCO3- and either Mg2+ or Mn2+. The inactivation rate follows saturation kinetics. Inactivation is slower in the presence of phospholactate or epoxymaleate, both of which are inhibitors of the enzyme, or dithiothreitol. Inactivation is completely prevented by the presence of lactate dehydrogenase and NADH, and 3-bromolactate is formed during this treatment. If the reaction is conducted by using HC18O3-, the inorganic phosphate produced contains 18O. This and other evidence indicate that phosphoenolpyruvate carboxylase catalyzes conversion of bromophosphoenolpyruvate into bromopyruvate by way of the usual carboxyphosphate-enolate intermediate, and bromopyruvate is the species responsible for enzyme inactivation. (Z)-3-fluorophosphoenolpyruvate is transformed by the enzyme into a 6:1 mixture of 3-fluoropyruvate and 3-fluorooxalacetate, presumably by the same mechanism. The enzyme is not inactivated during this treatment

  9. Reaction of phosphoenolpyruvate carboxylase with (Z)-3-bromophosphoenolpyruvate and (Z)-3-fluorophosphoenolpyruvate

    Energy Technology Data Exchange (ETDEWEB)

    Diaz, E.; O' Laughlin, J.T.; O' Leary, M.H.

    1988-02-23

    (Z)-3-Bromophosphoenolpyruvate inactivates phosphoenolpyruvate carboxylase from maize in the presence of HCO/sub 3//sup -/ and either Mg/sup 2 +/ or Mn/sup 2 +/. The inactivation rate follows saturation kinetics. Inactivation is slower in the presence of phospholactate or epoxymaleate, both of which are inhibitors of the enzyme, or dithiothreitol. Inactivation is completely prevented by the presence of lactate dehydrogenase and NADH, and 3-bromolactate is formed during this treatment. If the reaction is conducted by using HC/sup 18/O/sub 3//sup -/, the inorganic phosphate produced contains /sup 18/O. This and other evidence indicate that phosphoenolpyruvate carboxylase catalyzes conversion of bromophosphoenolpyruvate into bromopyruvate by way of the usual carboxyphosphate-enolate intermediate, and bromopyruvate is the species responsible for enzyme inactivation. (Z)-3-fluorophosphoenolpyruvate is transformed by the enzyme into a 6:1 mixture of 3-fluoropyruvate and 3-fluorooxalacetate, presumably by the same mechanism. The enzyme is not inactivated during this treatment.

  10. Inhibition of acetyl-CoA carboxylase by cystamine may mediate the hypotriglyceridemic activity of pantethine.

    Science.gov (United States)

    McCarty, M F

    2001-03-01

    Pantethine is a versatile and well-tolerated hypolipidemic agent whose efficacy in this regard appears to be mediated by its catabolic product cystamine, a nucleophile which avidly attacks disulfide groups. An overview of pantethine research suggests that the hypotriglyceridemic activity of pantethine reflects cystamine-mediated inhibition of the hepatic acetyl-CoA carboxylase, which can be expected to activate hepatic fatty acid oxidation. Inhibition of HMG-CoA reductase as well as a more distal enzyme in the cholesterol synthetic pathway may account for pantethine's hypocholesterolemic effects. If pantethine does indeed effectively inhibit hepatic acetyl-CoA carboxylase, it may have adjuvant utility in the hepatothermic therapy of obesity. As a safe and effective compound of natural origin, pantethine merits broader use in the management of hyperlipidemias. PMID:11359352

  11. Vitamin K-dependent carboxylase: possible role of the substrate "propeptide" as an intracellular recognition site.

    Science.gov (United States)

    Suttie, J W; Hoskins, J A; Engelke, J; Hopfgartner, A; Ehrlich, H; Bang, N U; Belagaje, R M; Schoner, B; Long, G L

    1987-01-01

    The liver microsomal vitamin K-dependent carboxylase catalyzes the posttranslational conversion of specific glutamate residues to gamma-carboxyglutamate residues in a limited number of proteins. A number of these proteins have been shown to contain a homologous basic amino acid-rich "propeptide" between the leader sequence and the amino terminus of the mature protein. Plasmids encoding protein C, a vitamin K-dependent protein, containing or lacking a propeptide region were constructed and the protein was expressed in Escherichia coli. The protein products were assayed as substrates in an in vitro vitamin K-dependent carboxylase system. Only proteins containing a propeptide region were substrates for the enzyme. These data support the hypothesis that this sequence of the primary gene product is an important recognition site for this processing enzyme. PMID:3543932

  12. Expression, purification and crystallization of an archaeal-type phosphoenolpyruvate carboxylase

    International Nuclear Information System (INIS)

    The expression, purification, crystallization and preliminary diffraction analysis of an archaeal-type phosphoenolpyruvate carboxylase are described. Complete highly redundant X-ray data have been measured from a crystal diffracting to 3.13 Å resolution. An archaeal-type phosphoenolpyruvate carboxylase (PepcA) from Clostridium perfringens has been expressed in Escherichia coli in a soluble form with an amino-terminal His tag. The recombinant protein is enzymatically active and two crystal forms have been obtained. Complete diffraction data extending to 3.13 Å resolution have been measured from a crystal soaked in KAu(CN)2, using radiation at a wavelength just above the Au LIII edge. The asymmetric unit contains two tetramers of PepcA

  13. Crystal Structures of Human and Staphylococcus aureus Pyruvate Carboxylase and Molecular Insights into the Carboxyltransfer Reaction

    Energy Technology Data Exchange (ETDEWEB)

    Xiang,S.; Tong, L.

    2008-01-01

    Pyruvate carboxylase (PC) catalyzes the biotin-dependent production of oxaloacetate and has important roles in gluconeogenesis, lipogenesis, insulin secretion and other cellular processes. PC contains the biotin carboxylase (BC), carboxyltransferase (CT) and biotin-carboxyl carrier protein (BCCP) domains. We report here the crystal structures at 2.8-Angstroms resolution of full-length PC from Staphylococcus aureus and the C-terminal region (missing only the BC domain) of human PC. A conserved tetrameric association is observed for both enzymes, and our structural and mutagenesis studies reveal a previously uncharacterized domain, the PC tetramerization (PT) domain, which is important for oligomerization. A BCCP domain is located in the active site of the CT domain, providing the first molecular insights into how biotin participates in the carboxyltransfer reaction. There are dramatic differences in domain positions in the monomer and the organization of the tetramer between these enzymes and the PC from Rhizobium etli.

  14. Crystal structure of the 500-kDa yeast acetyl-CoA carboxylase holoenzyme dimer.

    Science.gov (United States)

    Wei, Jia; Tong, Liang

    2015-10-29

    Acetyl-CoA carboxylase (ACC) has crucial roles in fatty acid metabolism and is an attractive target for drug discovery against diabetes, cancer and other diseases. Saccharomyces cerevisiae ACC (ScACC) is crucial for the production of very-long-chain fatty acids and the maintenance of the nuclear envelope. ACC contains biotin carboxylase (BC) and carboxyltransferase (CT) activities, and its biotin is linked covalently to the biotin carboxyl carrier protein (BCCP). Most eukaryotic ACCs are 250-kilodalton (kDa), multi-domain enzymes and function as homodimers and higher oligomers. They contain a unique, 80-kDa central region that shares no homology with other proteins. Although the structures of the BC, CT and BCCP domains and other biotin-dependent carboxylase holoenzymes are known, there is currently no structural information on the ACC holoenzyme. Here we report the crystal structure of the full-length, 500-kDa holoenzyme dimer of ScACC. The structure is remarkably different from that of the other biotin-dependent carboxylases. The central region contains five domains and is important for positioning the BC and CT domains for catalysis. The structure unexpectedly reveals a dimer of the BC domain and extensive conformational differences compared to the structure of the BC domain alone, which is a monomer. These structural changes reveal why the BC domain alone is catalytically inactive and define the molecular mechanism for the inhibition of eukaryotic ACC by the natural product soraphen A and by phosphorylation of a Ser residue just before the BC domain core in mammalian ACC. The BC and CT active sites are separated by 80 Å, and the entire BCCP domain must translocate during catalysis. PMID:26458104

  15. Vitamin K-dependent carboxylase: possible role of the substrate "propeptide" as an intracellular recognition site.

    OpenAIRE

    Suttie, J W; Hoskins, J A; Engelke, J; Hopfgartner, A; Ehrlich, H.; Bang, N U; Belagaje, R M; Schoner, B; Long, G L

    1987-01-01

    The liver microsomal vitamin K-dependent carboxylase catalyzes the posttranslational conversion of specific glutamate residues to gamma-carboxyglutamate residues in a limited number of proteins. A number of these proteins have been shown to contain a homologous basic amino acid-rich "propeptide" between the leader sequence and the amino terminus of the mature protein. Plasmids encoding protein C, a vitamin K-dependent protein, containing or lacking a propeptide region were constructed and the...

  16. Heterogeneity of holocarboxylase synthetase in patients with biotin-responsive multiple carboxylase deficiency.

    OpenAIRE

    Burri, B J; Sweetman, L.; Nyhan, W L

    1985-01-01

    Holocarboxylase synthetase activity has been determined in fibroblasts of seven patients with the neonatal form of biotin-responsive multiple carboxylase deficiency. The normal Km for biotin was 15 +/- 3 nmol/l, while in the patients the values ranged from 48 to 1,062 nmol/l. The mean maximum velocity was 27% of normal. Differences among the values obtained for the Km for biotin and the heat stability of holocarboxylase synthetase suggested that the patients studied represented at least four ...

  17. Deregulation of Feedback Inhibition of Phosphoenolpyruvate Carboxylase for Improved Lysine Production in Corynebacterium glutamicum

    OpenAIRE

    Chen, Zhen; Bommareddy, Rajesh Reddy; Frank, Doinita; Rappert, Sugima; Zeng, An-Ping

    2014-01-01

    Allosteric regulation of phosphoenolpyruvate carboxylase (PEPC) controls the metabolic flux distribution of anaplerotic pathways. In this study, the feedback inhibition of Corynebacterium glutamicum PEPC was rationally deregulated, and its effect on metabolic flux redistribution was evaluated. Based on rational protein design, six PEPC mutants were designed, and all of them showed significantly reduced sensitivity toward aspartate and malate inhibition. Introducing one of the point mutations ...

  18. Effect of Vitamin K-dependent Protein Precursor Propeptide, Vitamin K Hydroquinone, and Glutamate Substrate Binding on the Structure and Function of γ-Glutamyl Carboxylase*

    OpenAIRE

    Higgins-Gruber, Shannon L.; Mutucumarana, Vasantha P.; Lin, Pen-Jen; Jorgenson, James W.; Stafford, Darrel W.; Straight, David L.

    2010-01-01

    The γ-glutamyl carboxylase utilizes four substrates to catalyze carboxylation of certain glutamic acid residues in vitamin K-dependent proteins. How the enzyme brings the substrates together to promote catalysis is an important question in understanding the structure and function of this enzyme. The propeptide is the primary binding site of the vitamin K-dependent proteins to carboxylase. It is also an effector of carboxylase activity. We tested the hypothesis that binding of substrates cause...

  19. Calcium Promotes the Formation of Syntaxin 1 Mesoscale Domains through Phosphatidylinositol 4,5-Bisphosphate.

    Science.gov (United States)

    Milovanovic, Dragomir; Platen, Mitja; Junius, Meike; Diederichsen, Ulf; Schaap, Iwan A T; Honigmann, Alf; Jahn, Reinhard; van den Bogaart, Geert

    2016-04-01

    Phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) is a minor component of total plasma membrane lipids, but it has a substantial role in the regulation of many cellular functions, including exo- and endocytosis. Recently, it was shown that PI(4,5)P2and syntaxin 1, a SNARE protein that catalyzes regulated exocytosis, form domains in the plasma membrane that constitute recognition sites for vesicle docking. Also, calcium was shown to promote syntaxin 1 clustering in the plasma membrane, but the molecular mechanism was unknown. Here, using a combination of superresolution stimulated emission depletion microscopy, FRET, and atomic force microscopy, we show that Ca(2+)acts as a charge bridge that specifically and reversibly connects multiple syntaxin 1/PI(4,5)P2complexes into larger mesoscale domains. This transient reorganization of the plasma membrane by physiological Ca(2+)concentrations is likely to be important for Ca(2+)-regulated secretion. PMID:26884341

  20. Photochemical labeling of bovine pancreatic ribonuclease A with 8-azidoadenosine 3',5'-bisphosphate

    International Nuclear Information System (INIS)

    A simple method has been developed for the preparation of 5'-32P-labeled 8-azidoadenosine 3',5'-bisphosphate (p8N3Ap) for use in photoaffinity labeling studies. Irradiation of a complex between p8N3Ap and bovine pancreatic ribonuclease A (RNase A) with light of 300-350 nm led to the covalent attachment of the nucleotide to the enzyme. RNase A could also be labeled in the dark with prephotolyzed p8N3Ap. In either case, the nucleotide reacted with the same tryptic peptide, encompassing amino acids 67-85 of the protein. The site of labeling was determined to be either Thr-78 or Thr-82, both of which are close to or at the pyrimidine binding site of the enzyme. This result is consistent with recent nuclear magnetic resonance and X-ray studies which indicate that 8-substituted adenine nucleotides interact with the pyrimidine binding site of RNase A

  1. Structure of a rabbit muscle fructose-1, 6-bisphosphate aldolase A dimer variant

    Energy Technology Data Exchange (ETDEWEB)

    Sherawat, Manashi [Department of Physiology and Biophysics, Boston University School of Medicine, 715 Albany Street, Boston, MA 02118-2394 (United States); Tolan, Dean R., E-mail: tolan@bu.edu [Department of Biology, Boston University, 5 Cummington Street, Boston, MA 02215 (United States); Allen, Karen N., E-mail: tolan@bu.edu [Department of Physiology and Biophysics, Boston University School of Medicine, 715 Albany Street, Boston, MA 02118-2394 (United States)

    2008-05-01

    The X-ray crystallographic structure of a dimer variant of fructose-1, 6-bisphosphate aldolase demonstrates a stable oligomer that mirrors half of the native tetramer. The presence of product demonstrates that this is an active form. Fructose-1, 6-bisphosphate aldolase (aldolase) is an essential enzyme in glycolysis and gluconeogenesis. In addition to this primary function, aldolase is also known to bind to a variety of other proteins, a property that may allow it to perform ‘moonlighting’ roles in the cell. Although monomeric and dimeric aldolases possess full catalytic activity, the enzyme occurs as an unusually stable tetramer, suggesting a possible link between the oligomeric state and these noncatalytic cellular roles. Here, the first high-resolution X-ray crystal structure of rabbit muscle D128V aldolase, a dimeric form of aldolase mimicking the clinically important D128G mutation in humans associated with hemolytic anemia, is presented. The structure of the dimer was determined to 1.7 Å resolution with the product DHAP bound in the active site. The turnover of substrate to produce the product ligand demonstrates the retention of catalytic activity by the dimeric aldolase. The D128V mutation causes aldolase to lose intermolecular contacts with the neighboring subunit at one of the two interfaces of the tetramer. The tertiary structure of the dimer does not significantly differ from the structure of half of the tetramer. Analytical ultracentrifugation confirms the occurrence of the enzyme as a dimer in solution. The highly stable structure of aldolase with an independent active site is consistent with a model in which aldolase has evolved as a multimeric scaffold to perform other noncatalytic functions.

  2. Structure of a rabbit muscle fructose-1, 6-bisphosphate aldolase A dimer variant

    International Nuclear Information System (INIS)

    The X-ray crystallographic structure of a dimer variant of fructose-1, 6-bisphosphate aldolase demonstrates a stable oligomer that mirrors half of the native tetramer. The presence of product demonstrates that this is an active form. Fructose-1, 6-bisphosphate aldolase (aldolase) is an essential enzyme in glycolysis and gluconeogenesis. In addition to this primary function, aldolase is also known to bind to a variety of other proteins, a property that may allow it to perform ‘moonlighting’ roles in the cell. Although monomeric and dimeric aldolases possess full catalytic activity, the enzyme occurs as an unusually stable tetramer, suggesting a possible link between the oligomeric state and these noncatalytic cellular roles. Here, the first high-resolution X-ray crystal structure of rabbit muscle D128V aldolase, a dimeric form of aldolase mimicking the clinically important D128G mutation in humans associated with hemolytic anemia, is presented. The structure of the dimer was determined to 1.7 Å resolution with the product DHAP bound in the active site. The turnover of substrate to produce the product ligand demonstrates the retention of catalytic activity by the dimeric aldolase. The D128V mutation causes aldolase to lose intermolecular contacts with the neighboring subunit at one of the two interfaces of the tetramer. The tertiary structure of the dimer does not significantly differ from the structure of half of the tetramer. Analytical ultracentrifugation confirms the occurrence of the enzyme as a dimer in solution. The highly stable structure of aldolase with an independent active site is consistent with a model in which aldolase has evolved as a multimeric scaffold to perform other noncatalytic functions

  3. Solar ultraviolet radiation affects the activity of ribulose-1, 5-bisphosphate carboxylase-oxygenase and the composition of photosynthetic and xanthophyll cycle pigments in the intertidal green alga Ulva lactuca L.

    NARCIS (Netherlands)

    Bischof, K; Krabs, G; Wiencke, C; Hanelt, D

    2002-01-01

    The effect of solar UV radiation on the physiology of the intertidal green macroalga Ulva lactuca L. was investigated. A natural Ulm community at the shore of Helgoland was covered with screening foils, excluding UV-B or UV-B + UV-A from the solar spectrum. In the sampled material, changes in the ac

  4. CO2 assimilation, ribulose-1,5-bisphosphate carboxylase/oxygenase, carbohydrates and photosynthetic electron transport probed by the JIP-test, of tea leaves in response to phosphorus supply

    Directory of Open Access Journals (Sweden)

    Chen Rong-Bing

    2009-04-01

    Full Text Available Abstract Background Although the effects of P deficiency on tea (Camellia sinensis (L. O. Kuntze growth, P uptake and utilization as well as leaf gas exchange and Chl a fluorescence have been investigated, very little is known about the effects of P deficiency on photosynthetic electron transport, photosynthetic enzymes and carbohydrates of tea leaves. In this study, own-rooted 10-month-old tea trees were supplied three times weekly for 17 weeks with 500 mL of nutrient solution at a P concentration of 0, 40, 80, 160, 400 or 1000 μM. This objective of this study was to determine how P deficiency affects CO2 assimilation, Rubisco, carbohydrates and photosynthetic electron transport in tea leaves to understand the mechanism by which P deficiency leads to a decrease in CO2 assimilation. Results Both root and shoot dry weight increased as P supply increased from 0 to 160 μM, then remained unchanged. P-deficient leaves from 0 to 80 μM P-treated trees showed decreased CO2 assimilation and stomatal conductance, but increased intercellular CO2 concentration. Both initial and total Rubisco activity, contents of Chl and total soluble protein in P-deficient leaves decreased to a lesser extent than CO2 assimilation. Contents of sucrose and starch were decreased in P-deficient leaves, whereas contents of glucose and fructose did not change significantly except for a significant increase in the lowest P leaves. OJIP transients from P-deficient leaves displayed a rise at the O-step and a depression at the P-step, accompanied by two new steps at about 150 μs (L-step and at about 300 μs (K-step. RC/CSo, TRo/ABS (or Fv/Fm, ETo/ABS, REo/ABS, maximum amplitude of IP phase, PIabs and PItot, abs were decreased in P-deficient leaves, while VJ, VI and dissipated energy were increased. Conclusion P deficiency decreased photosynthetic electron transport capacity by impairing the whole electron transport chain from the PSII donor side up to the PSI, thus decreasing ATP content which limits RuBP regeneration, and hence, the rate of CO2 assimilation. Energy dissipation is enhanced to protect P-deficient leaves from photo-oxidative damage in high light.

  5. Wheat cytosolic acetyl-CoA carboxylase complements an ACC1 null mutation in yeast

    OpenAIRE

    Joachimiak, M.; Tevzadze, G.; Podkowinski, J; Haselkorn, R.; Gornicki, P.

    1997-01-01

    Spores harboring an ACC1 deletion derived from a diploid Saccharomyces cerevisiae strain, in which one copy of the entire ACC1 gene is replaced with a LEU2 cassette, fail to grow. A chimeric gene consisting of the yeast GAL10 promoter, yeast ACC1 leader, wheat cytosolic acetyl-CoA carboxylase (ACCase) cDNA, and yeast ACC1 3′ tail was used to complement a yeast ACC1 mutation. The complementation demonstrates that active wheat ACCase can be produced in yeast. At low concentrations of galactose,...

  6. Attempts to apply affinity labeling techniques to ribulosebisphosphate carboxylase/oxygenase. [Comparison of spinach leaf and Rhodospirillum rubrum

    Energy Technology Data Exchange (ETDEWEB)

    Hartman, F. C.; Norton, I. L.; Stringer, C. D.; Schloss, J. V.

    1978-01-01

    Studies on carboxylases/oxygenases from different species may be necessary to confirm that a residue implicated as essential is indeed an active-site component. To provide an especially stringent test case for the identification of species invariant structural features the enzymes from two phylogenetically distant species, spinach and Rhodospirillum rubrum, were compared. To date, the reactions of Br-butanone-P/sub 2/ and BrAcNHEtOP with the spinach enayme have been rather thoroughly characterized; only preliminary experiments have been completed with the R. rubrum enzyme. Both enzymes were isolated and assayed for carboxylase activity (spectrophotometrically or /sup 14/CO/sub 2/-fixation) and for oxygenase activity.

  7. A Symmetrical Tetramer for S. aureus Pyruvate Carboxylase in Complex with Coenzyme A

    Energy Technology Data Exchange (ETDEWEB)

    Yu, L.; Xiang, S; Lasso, G; Gil, D; Valle, M; Tong, L

    2009-01-01

    Pyruvate carboxylase (PC) is a conserved metabolic enzyme with important cellular functions. We report crystallographic and cryo-electron microscopy (EM) studies of Staphylococcus aureus PC (SaPC) in complex with acetyl-CoA, an allosteric activator, and mutagenesis, biochemical, and structural studies of the biotin binding site of its carboxyltransferase (CT) domain. The disease-causing A610T mutation abolishes catalytic activity by blocking biotin binding to the CT active site, and Thr908 might play a catalytic role in the CT reaction. The crystal structure of SaPC in complex with CoA reveals a symmetrical tetramer, with one CoA molecule bound to each monomer, and cryo-EM studies confirm the symmetrical nature of the tetramer. These observations are in sharp contrast to the highly asymmetrical tetramer of Rhizobium etli PC in complex with ethyl-CoA. Our structural information suggests that acetyl-CoA promotes a conformation for the dimer of the biotin carboxylase domain of PC that might be catalytically more competent.

  8. Oxygen requirement and inhibition of C4 photosynthesis

    OpenAIRE

    Maroco, João; Ku, Maurice S. B.; Peter J. Lea; Dever, Louisa V.; Leegood, Richard C.; Furbank, Robert T.; Edwards, Gerald E.

    1998-01-01

    The basis for O2 sensitivity of C4 photosynthesis was evaluated using a C4-cycle-limited mutant of Amaranthus edulis (a phosphoenolpyruvate carboxylase-deficient mutant), and a C3-cyclelimited transformant of Flaveria bidentis (an antisense ribulose-1,5- bisphosphate carboxylase/oxygenase [Rubisco] small subunit transformant). Data obtained with the C4-cycle-limited mutant showed that atmospheric levels of O2 (20 kPa) caused increased inhibition of photosynthesis as a res...

  9. Insights into the carboxyltransferase reaction of pyruvate carboxylase from the structures of bound product and intermediate analogs.

    Science.gov (United States)

    Lietzan, Adam D; St Maurice, Martin

    2013-11-15

    Pyruvate carboxylase (PC) is a biotin-dependent enzyme that catalyzes the MgATP- and bicarbonate-dependent carboxylation of pyruvate to oxaloacetate, an important anaplerotic reaction in central metabolism. The carboxyltransferase (CT) domain of PC catalyzes the transfer of a carboxyl group from carboxybiotin to the accepting substrate, pyruvate. It has been hypothesized that the reactive enolpyruvate intermediate is stabilized through a bidentate interaction with the metal ion in the CT domain active site. Whereas bidentate ligands are commonly observed in enzymes catalyzing reactions proceeding through an enolpyruvate intermediate, no bidentate interaction has yet been observed in the CT domain of PC. Here, we report three X-ray crystal structures of the Rhizobium etli PC CT domain with the bound inhibitors oxalate, 3-hydroxypyruvate, and 3-bromopyruvate. Oxalate, a stereoelectronic mimic of the enolpyruvate intermediate, does not interact directly with the metal ion. Instead, oxalate is buried in a pocket formed by several positively charged amino acid residues and the metal ion. Furthermore, both 3-hydroxypyruvate and 3-bromopyruvate, analogs of the reaction product oxaloacetate, bind in an identical manner to oxalate suggesting that the substrate maintains its orientation in the active site throughout catalysis. Together, these structures indicate that the substrates, products and intermediates in the PC-catalyzed reaction are not oriented in the active site as previously assumed. The absence of a bidentate interaction with the active site metal appears to be a unique mechanistic feature among the small group of biotin-dependent enzymes that act on α-keto acid substrates. PMID:24157795

  10. Structural basis for the bifunctionality of fructose-1,6-bisphosphate aldolase/phosphatase.

    Science.gov (United States)

    Fushinobu, Shinya; Nishimasu, Hiroshi; Hattori, Daiki; Song, Hyun-Jin; Wakagi, Takayoshi

    2011-10-27

    Enzymes catalyse specific reactions and are essential for maintaining life. Although some are referred to as being bifunctional, they consist of either two distinct catalytic domains or a single domain that displays promiscuous substrate specificity. Thus, one enzyme active site is generally responsible for one biochemical reaction. In contrast to this conventional concept, archaeal fructose-1,6-bisphosphate (FBP) aldolase/phosphatase (FBPA/P) consists of a single catalytic domain, but catalyses two chemically distinct reactions of gluconeogenesis: (1) the reversible aldol condensation of dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (GA3P) to FBP; (2) the dephosphorylation of FBP to fructose-6-phosphate (F6P). Thus, FBPA/P is fundamentally different from ordinary enzymes whose active sites are responsible for a specific reaction. However, the molecular mechanism by which FBPA/P achieves its unusual bifunctionality remains unknown. Here we report the crystal structure of FBPA/P at 1.5-Å resolution in the aldolase form, where a critical lysine residue forms a Schiff base with DHAP. A structural comparison of the aldolase form with a previously determined phosphatase form revealed a dramatic conformational change in the active site, demonstrating that FBPA/P metamorphoses its active-site architecture to exhibit dual activities. Thus, our findings expand the conventional concept that one enzyme catalyses one biochemical reaction. PMID:21983966

  11. Active-site remodelling in the bifunctional fructose-1,6-bisphosphate aldolase/phosphatase.

    Science.gov (United States)

    Du, Juan; Say, Rafael F; Lü, Wei; Fuchs, Georg; Einsle, Oliver

    2011-10-27

    Fructose-1,6-bisphosphate (FBP) aldolase/phosphatase is a bifunctional, thermostable enzyme that catalyses two subsequent steps in gluconeogenesis in most archaea and in deeply branching bacterial lineages. It mediates the aldol condensation of heat-labile dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (GAP) to FBP, as well as the subsequent, irreversible hydrolysis of the product to yield the stable fructose-6-phosphate (F6P) and inorganic phosphate; no reaction intermediates are released. Here we present a series of structural snapshots of the reaction that reveal a substantial remodelling of the active site through the movement of loop regions that create different catalytic functionalities at the same location. We have solved the three-dimensional structures of FBP aldolase/phosphatase from thermophilic Thermoproteus neutrophilus in a ligand-free state as well as in complex with the substrates DHAP and FBP and the product F6P to resolutions up to 1.3 Å. In conjunction with mutagenesis data, this pinpoints the residues required for the two reaction steps and shows that the sequential binding of additional Mg(2+) cations reversibly facilitates the reaction. FBP aldolase/phosphatase is an ancestral gluconeogenic enzyme optimized for high ambient temperatures, and our work resolves how consecutive structural rearrangements reorganize the catalytic centre of the protein to carry out two canonical reactions in a very non-canonical type of bifunctionality. PMID:21983965

  12. NMR studies of fructose-1,6-bisphosphate aldolase from E. coli

    International Nuclear Information System (INIS)

    Previous NMR studies of intact E. coli showed that during steady state anaerobic catabolism of glucose the main glycolytic intermediate detectable in these cells is fructose-1,6-bisphosphate (FBP), levels of which remain constant while the levels of glucose, lactate and succinate vary considerably. Upon feeding these cells glucose labeled with 13C at the C1 or C6 position, the level of scrambling of label between the C1 and C6 positions of FBP was low suggesting that the FBP-aldolase reaction is far from equilibrium. In order to account for these observations, a study was undertaken on FBP-aldolase from this organism. This enzyme is a dimeric Zn++ metalloenzyme with a M/sub r/ of 80,000. It was purified in gram quantities from an overproducer strain and was characterized by standard biochemical techniques prior to the NMR studies. 13C NMR experiments were conducted using [2-13C]dihydroxyacetone phosphate (DHAP) and [2,5-13C]fructose-1,6-biphosphate (FBP). Since these substrates can exist in solution in a number of interconvertible forms, the initial experiments determined the relative amounts of these forms and the rates of their interconversion. Subsequently, NMR experiments with the purified enzyme were conducted. Based upon these results, the author concludes that in E. coli the FBP-alkolase reaction appears to be the rate limiting step of anaerobic glycolysis

  13. Phosphatidylinositol 4,5-bisphosphate alters pharmacological selectivity for epilepsy-causing KCNQ potassium channels

    Science.gov (United States)

    Zhou, Pingzheng; Yu, Haibo; Gu, Min; Nan, Fa-jun; Gao, Zhaobing; Li, Min

    2013-01-01

    Pharmacological augmentation of neuronal KCNQ muscarinic (M) currents by drugs such as retigabine (RTG) represents a first-in-class therapeutic to treat certain hyperexcitatory diseases by dampening neuronal firing. Whereas all five potassium channel subtypes (KCNQ1–KCNQ5) are found in the nervous system, KCNQ2 and KCNQ3 are the primary players that mediate M currents. We investigated the plasticity of subtype selectivity by two M current effective drugs, retigabine and zinc pyrithione (ZnPy). Retigabine is more effective on KCNQ3 than KCNQ2, whereas ZnPy is more effective on KCNQ2 with no detectable effect on KCNQ3. In neurons, activation of muscarinic receptor signaling desensitizes effects by retigabine but not ZnPy. Importantly, reduction of phosphatidylinositol 4,5-bisphosphate (PIP2) causes KCNQ3 to become sensitive to ZnPy but lose sensitivity to retigabine. The dynamic shift of pharmacological selectivity caused by PIP2 may be induced orthogonally by voltage-sensitive phosphatase, or conversely, abolished by mutating a PIP2 site within the S4–S5 linker of KCNQ3. Therefore, whereas drug-channel binding is a prerequisite, the drug selectivity on M current is dynamic and may be regulated by receptor signaling pathways via PIP2. PMID:23650395

  14. Novel Phosphotidylinositol 4,5-Bisphosphate Binding Sites on Focal Adhesion Kinase.

    Directory of Open Access Journals (Sweden)

    Jun Feng

    Full Text Available Focal adhesion kinase (FAK is a protein tyrosine kinase that is ubiquitously expressed, recruited to focal adhesions, and engages in a variety of cellular signaling pathways. Diverse cellular responses, such as cell migration, proliferation, and survival, are regulated by FAK. Prior to activation, FAK adopts an autoinhibited conformation in which the FERM domain binds the kinase domain, blocking access to the activation loop and substrate binding site. Activation of FAK occurs through conformational change, and acidic phospholipids such as phosphatidylinositol 4,5-bisphosphate (PIP2 are known to facilitate this process. PIP2 binding alters the autoinhibited conformation of the FERM and kinase domains and subsequently exposes the activation loop to phosphorylation. However, the detailed molecular mechanism of PIP2 binding and its role in FAK activation remain unclear. In this study, we conducted coarse-grained molecular dynamics simulations to investigate the binding of FAK to PIP2. Our simulations identified novel areas of basic residues in the kinase domain of FAK that potentially undergo transient binding to PIP2 through electrostatic attractions. Our investigation provides a molecular picture of PIP2-initiated FAK activation and introduces promising new pathways for future studies of FAK regulation.

  15. Structure of a Rabbit Muscle Fructose-1,6-Bisphosphate Aldolase A Dimer Variant

    Energy Technology Data Exchange (ETDEWEB)

    Sherawat,M.; Tolan, D.; Allen, K.

    2008-01-01

    Fructose-1,6-bisphosphate aldolase (aldolase) is an essential enzyme in glycolysis and gluconeogenesis. In addition to this primary function, aldolase is also known to bind to a variety of other proteins, a property that may allow it to perform 'moonlighting' roles in the cell. Although monomeric and dimeric aldolases possess full catalytic activity, the enzyme occurs as an unusually stable tetramer, suggesting a possible link between the oligomeric state and these noncatalytic cellular roles. Here, the first high-resolution X-ray crystal structure of rabbit muscle D128V aldolase, a dimeric form of aldolase mimicking the clinically important D128G mutation in humans associated with hemolytic anemia, is presented. The structure of the dimer was determined to 1.7 Angstroms resolution with the product DHAP bound in the active site. The turnover of substrate to produce the product ligand demonstrates the retention of catalytic activity by the dimeric aldolase. The D128V mutation causes aldolase to lose intermolecular contacts with the neighboring subunit at one of the two interfaces of the tetramer. The tertiary structure of the dimer does not significantly differ from the structure of half of the tetramer. Analytical ultracentrifugation confirms the occurrence of the enzyme as a dimer in solution. The highly stable structure of aldolase with an independent active site is consistent with a model in which aldolase has evolved as a multimeric scaffold to perform other noncatalytic functions.

  16. Resolving the Activation Site of PositiveRegulators in Plant PhosphoenolpyruvateCarboxylase

    Institute of Scientific and Technical Information of China (English)

    2014-01-01

    Dear Editor, Phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) islocated at an important branch point in the carbohydratemetabolism of plants. The enzyme is a homotetramer andcatalyzes the addition of bicarbonate to phosphoenolpyru-vate (PEP) to form oxaloacetate and phosphate. PEPC isregulated by metabolites and phosphorylation. AIIostericfeedback inhibition is mainly regulated by L-malate andL-aspartate which bind to a site separated from the activecenter (Kai et al., 1999; Paulus et al., 2013). Structure analy-sis of PEPC from Escherichia coli (Kai et al., 1999; Matsumuraet al., 2002), Zea rnays (Matsumura et al., 2002), Flaveria trin-ervia, and F. pringlei (Paulus et al., 2013) revealed that thesubstrate PEP and the feedback inhibitors bind to separatesites within each monomer.

  17. Is L-Carnitine Supplementation Beneficial in 3-Methylcrotonyl-CoA Carboxylase Deficiency?

    DEFF Research Database (Denmark)

    Thomsen, Jákup Andreas; Lund, Allan Meldgaard; Olesen, Jess Have; Mohr, Magni; Rasmussen, Jan

    2015-01-01

    Background: 3-Methylcrotonyl-CoA carboxylase deficiency (3-MCCd) is an autosomal recessive disorder in the catabolism of leucine. In the present study, we investigated the current and prior medical condition of patients with 3-MCCd in the Faroe Islands and their carnitine levels in blood, urine and...... muscle tissue with and without L-carnitine supplementation to evaluate the current treatment strategy of not recommending L-carnitine supplementation to Faroese 3-MCCd patients. Methods: Blood and urine samples and muscle biopsies were collected from patients at inclusion and at 3 months. Eight patients...... received L-carnitine supplementation when recruited; five did not. Included patients who received supplementation were asked to stop L-carnitine, the others were asked to initiate L-carnitine supplementation during the study. Symptoms were determined by review of hospital medical records and questionnaires...

  18. Acetyl CoA Carboxylase 2 Is Dispensable for CD8+ T Cell Responses.

    Directory of Open Access Journals (Sweden)

    Jang Eun Lee

    Full Text Available Differentiation of T cells is closely associated with dynamic changes in nutrient and energy metabolism. However, the extent to which specific metabolic pathways and molecular components are determinative of CD8+ T cell fate remains unclear. It has been previously established in various tissues that acetyl CoA carboxylase 2 (ACC2 regulates fatty acid oxidation (FAO by inhibiting carnitine palmitoyltransferase 1 (CPT1, a rate-limiting enzyme of FAO in mitochondria. Here, we explore the cell-intrinsic role of ACC2 in T cell immunity in response to infections. We report here that ACC2 deficiency results in a marginal increase of cellular FAO in CD8+ T cells, but does not appear to influence antigen-specific effector and memory CD8+ T cell responses during infection with listeria or lymphocytic choriomeningitis virus. These results suggest that ACC2 is dispensable for CD8+ T cell responses.

  19. Novel Mutations in the PC Gene in Patients with Type B Pyruvate Carboxylase Deficiency

    DEFF Research Database (Denmark)

    Ostergaard, Elsebet; Duno, Morten; Møller, Lisbeth Birk;

    2013-01-01

    We have investigated seven patients with the type B form of pyruvate carboxylase (PC) deficiency. Mutation analysis revealed eight mutations, all novel. In a patient with exon skipping on cDNA analysis, we identified a homozygous mutation located in a potential branch point sequence, the first...... possible branch point mutation in PC. Two patients were homozygous for missense mutations (with normal protein amounts on western blot analysis), and two patients were homozygous for nonsense mutations. In addition, a duplication of one base pair was found in a patient who also harboured a splice site...... mutation. Another splice site mutation led to the activation of a cryptic splice site, shown by cDNA analysis.All patients reported until now with at least one missense mutation have had the milder type A form of PC deficiency. We thus report for the first time two patients with homozygous missense...

  20. Structural and Biochemical Studies on the Regulation of Biotin Carboxylase by Substrate Inhibition and Dimerization

    Energy Technology Data Exchange (ETDEWEB)

    Chou, Chi-Yuan; Tong, Liang (Columbia)

    2012-06-19

    Biotin carboxylase (BC) activity is shared among biotin-dependent carboxylases and catalyzes the Mg-ATP-dependent carboxylation of biotin using bicarbonate as the CO{sub 2} donor. BC has been studied extensively over the years by structural, kinetic, and mutagenesis analyses. Here we report three new crystal structures of Escherichia coli BC at up to 1.9 {angstrom} resolution, complexed with different ligands. Two structures are wild-type BC in complex with two ADP molecules and two Ca{sup 2+} ions or two ADP molecules and one Mg{sup 2+} ion. One ADP molecule is in the position normally taken by the ATP substrate, whereas the other ADP molecule occupies the binding sites of bicarbonate and biotin. One Ca{sup 2+} ion and the Mg{sup 2+} ion are associated with the ADP molecule in the active site, and the other Ca{sup 2+} ion is coordinated by Glu-87, Glu-288, and Asn-290. Our kinetic studies confirm that ATP shows substrate inhibition and that this inhibition is competitive against bicarbonate. The third structure is on the R16E mutant in complex with bicarbonate and Mg-ADP. Arg-16 is located near the dimer interface. The R16E mutant has only a 2-fold loss in catalytic activity compared with the wild-type enzyme. Analytical ultracentrifugation experiments showed that the mutation significantly destabilized the dimer, although the presence of substrates can induce dimer formation. The binding modes of bicarbonate and Mg-ADP are essentially the same as those to the wild-type enzyme. However, the mutation greatly disrupted the dimer interface and caused a large re-organization of the dimer. The structures of these new complexes have implications for the catalysis by BC.

  1. Inhibition of E. coli P-enolpyruvate carboxylase by P-enol-3-bromopyruvate

    International Nuclear Information System (INIS)

    The generality of the mechanism based inhibition of P-enolpyruvate carboxylases (PEPCase) by P-enol-3-bromopyruvate (BrPEP) was tested by measuring its effects on the allosterically regulated enzyme from E. coli. In the presence of 1mM Mn2+, BrPEP appears to be a competitive inhibitor (K/sub i/ = 0.0087mM) of PEPCase. Incubation of 0.005mM PEPCase with 0.5mM (or 1.0mM)BrPEP along with H14CO3- and Mn2+, yielded, upon reduction with NaBH4, a protein containing radioactivity in an amount approximately proportional to that expected from the loss of catalytic activity. At both a 25- and a 50-fold excess (0.5mM and 1.0mM, respectively) of BrPEP to PEPCase subunits, first order loss of activity occurred with k values of 5.24 x 10-3 min-1 and 1.03 x 10-2 min-1, respectively. At the lower concentration of BrPEP the inactivation process appeared to be reversible after 40 min with no further inhibition occurring even up to two hours of incubation. At the higher concentration of BrPEP, the rate of inhibition slowed dramatically after 50 min and appeared insignificant over the next hour. These data suggest that BrPEP irreversibly inactivates the E. coli PEP carboxylase, but that there may be considerable dissociation of the product, Br-oxaloacetate, before irreversible binding occurs, and that the reduced rate of inactivation may be due to depletion of BrPEP

  2. Phosphatidylinositol 4,5-bisphosphate competitively inhibits phorbol ester binding to protein kinase C

    International Nuclear Information System (INIS)

    Calcium phospholipid dependent protein kinase C (PKC) is activated by diacylglycerol (DG) and by phorbol esters and is recognized to be the phorbol ester receptor of cells; DG displaces phorbol ester competitively from PKC. A phospholipid, phosphatidylinositol 4,5-bisphosphate (PIP2), can also activate PKC in the presence of phosphatidylserine (PS) and Ca2+ with a KPIP2 of 0.04 mol %. Preliminary experiments have suggested a common binding site for PIP2 and DG on PKC. Here, the authors investigate the effect of PIP2 on phorbol ester binding to PKC in a mixed micellar assay. In the presence of 20 mol % PS, PIP2 inhibited specific binding of [3H]phorbol 12,13-dibutyrate (PDBu) in a dose-dependent fashion up to 85% at 1 mol %. Inhibition of binding was more pronounced with PIP2 than with DG. Scatchard analysis indicated that the decrease in binding of PDBu in the presence of PIP2 is the result of an altered affinity for the phorbol ester rather than of a change in maximal binding. The plot of apparent dissociation constants (Kd') against PIP2 concentration was linear over a range of 0.01-1 mol % with a Ki of 0.043 mol % and confirmed the competitive nature of inhibition between PDBu and PIP2. Competition between PIP2 and phorbol ester could be determined in a liposomal assay system also. These results indicate that PIP2, DG, and phorbol ester all compete for the same activator-receiving region on the regulatory moiety of protein kinase C, and they lend support to the suggestion that PIP2 is a primary activator of the enzyme

  3. Fructose 1,6-Bisphosphate Aldolase, a Novel Immunogenic Surface Protein on Listeria Species

    Science.gov (United States)

    Conceição, Fabricio Rochedo; Hust, Michael; Mendonça, Karla Sequeira; Moreira, Ângela Nunes; França, Rodrigo Correa; da Silva, Wladimir Padilha; Aleixo, José Antonio G.

    2016-01-01

    Listeria monocytogenes is a ubiquitous food-borne pathogen, and its presence in food or production facilities highlights the importance of surveillance. Increased understanding of the surface exposed antigens on Listeria would provide potential diagnostic and therapeutic targets. In the present work, using mass spectrometry and genetic cloning, we show that fructose-1,6-bisphosphate aldolase (FBA) class II in Listeria species is the antigen target of the previously described mAb-3F8. Western and dot blot assays confirmed that the mAb-3F8 could distinguish all tested Listeria species from close-related bacteria. Localization studies indicated that FBA is present in every fraction of Listeria cells, including supernatant and the cell wall, setting Listeria spp. as one of the few bacteria described to have this protein on their cell surface. Epitope mapping using ORFeome display and a peptide membrane revealed a 14-amino acid peptide as the potential mAb-3F8 epitope. The target epitope in FBA allowed distinguishing Listeria spp. from closely-related bacteria, and was identified as part of the active site in the dimeric enzyme. However, its function in cell surface seems not to be host cell adhesion-related. Western and dot blot assays further demonstrated that mAb-3F8 together with anti-InlA mAb-2D12 could differentiate pathogenic from non-pathogenic Listeria isolated from artificially contaminated cheese. In summary, we report FBA as a novel immunogenic surface target useful for the detection of Listeria genus. PMID:27489951

  4. Response of fructose-2,6-bisphosphate to environmental changes. Effect of low temperature in winter and spring wheat

    OpenAIRE

    Van Praag, Esther; Degli Agosti, Robert

    1997-01-01

    Response of fructose-2,6-bisphosphate to environmental changes. Effect of low temperature in winter and spring wheat - Three day old, etiolated, wheat seedlings were affected differently according to the duration of exposure to a non-freezing low temperature (5°C). Indeed, growth was only reduced after a 45 min or longer treatment. Germs exposed to a 120 min cold period were half the size 24 h after the cold treatment; in comparison to the room temperature (20°C) exposed seedlings. ln respect...

  5. Molecular cloning and characterization of the cDNA coding for the biotin-containing subunit of 3-methylcrotonoyl-CoA carboxylase: identification of the biotin carboxylase and biotin-carrier domains.

    OpenAIRE

    Song, J.; Wurtele, E S; Nikolau, B J

    1994-01-01

    Soybean genomic clones were isolated based on hybridization to probes that code for the conserved biotinylation domain of biotin-containing enzymes. The corresponding cDNA was isolated and expressed in Escherichia coli through fusion to the bacterial trpE gene. The resulting chimeric protein was biotinylated in E. coli. Antibodies raised against the chimeric protein reacted specifically with an 85-kDa biotin-containing polypeptide from soybean and inhibited 3-methylcrotonoyl-CoA carboxylase (...

  6. Evolution and Functional Diversification of Fructose Bisphosphate Aldolase Genes in Photosynthetic Marine Diatoms

    Science.gov (United States)

    Allen, Andrew E.; Moustafa, Ahmed; Montsant, Anton; Eckert, Angelika; Kroth, Peter G.; Bowler, Chris

    2012-01-01

    Diatoms and other chlorophyll-c containing, or chromalveolate, algae are among the most productive and diverse phytoplankton in the ocean. Evolutionarily, chlorophyll-c algae are linked through common, although not necessarily monophyletic, acquisition of plastid endosymbionts of red as well as most likely green algal origin. There is also strong evidence for a relatively high level of lineage-specific bacterial gene acquisition within chromalveolates. Therefore, analyses of gene content and derivation in chromalveolate taxa have indicated particularly diverse origins of their overall gene repertoire. As a single group of functionally related enzymes spanning two distinct gene families, fructose 1,6-bisphosphate aldolases (FBAs) illustrate the influence on core biochemical pathways of specific evolutionary associations among diatoms and other chromalveolates with various plastid-bearing and bacterial endosymbionts. Protein localization and activity, gene expression, and phylogenetic analyses indicate that the pennate diatom Phaeodactylum tricornutum contains five FBA genes with very little overall functional overlap. Three P. tricornutum FBAs, one class I and two class II, are plastid localized, and each appears to have a distinct evolutionary origin as well as function. Class I plastid FBA appears to have been acquired by chromalveolates from a red algal endosymbiont, whereas one copy of class II plastid FBA is likely to have originated from an ancient green algal endosymbiont. The other copy appears to be the result of a chromalveolate-specific gene duplication. Plastid FBA I and chromalveolate-specific class II plastid FBA are localized in the pyrenoid region of the chloroplast where they are associated with β-carbonic anhydrase, which is known to play a significant role in regulation of the diatom carbon concentrating mechanism. The two pyrenoid-associated FBAs are distinguished by contrasting gene expression profiles under nutrient limiting compared with

  7. Propeptide and glutamate-containing substrates bound to the vitamin K-dependent carboxylase convert its vitamin K epoxidase function from an inactive to an active state

    OpenAIRE

    Sugiura, Isamu; Furie, Bruce; Walsh, Christopher T.; Furie, Barbara C.

    1997-01-01

    The vitamin K-dependent γ-glutamyl carboxylase catalyzes the posttranslational conversion of glutamic acid to γ-carboxyglutamic acid in precursor proteins containing the γ-carboxylation recognition site (γ-CRS). During this reaction, glutamic acid is converted to γ-carboxyglutamic acid while vitamin KH2 is converted to vitamin K 2,3-epoxide. Recombinant bovine carboxylase was purified free of γ-CRS-containing propeptide and endogenous substrate in a single-step immunoaffinity procedure. We sh...

  8. /sup 13/C nuclear magnetic resonance study of the CO/sub 2/ activation of ribulosebisphosphate carboxylase from Rhodospirillum rubrum

    Energy Technology Data Exchange (ETDEWEB)

    O' Leary, M.H. (Univ. of Wisconsin, Madison); Joworski, R.J.; Hartman, F.C.

    1979-02-01

    Ribulosebisphosphate carboxylase (3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39) from Rhodospirillum rubrum is activated by CO/sub 2/ and Mg/sup 2 +/. /sup 13/C NMR spectra were determined for the unactivated enzyme and for enzyme that had been activated by /sup 13/CO/sub 2/ and Mg/sup 2 +/. In addition to the expected resonance for H/sup 13/CO/sub 3//sup -//CO/sub 3//sup 2 -/ at 161.8 ppM downfield from tetramethylsilane, the spectrum of the activated enzyme shows a broad resonance at 164.9 ppM. Analogy with previous NMR studies of /sup 13/CO/sub 2/ binding to hemoglobin suggests that the CO/sub 2/ activation of ribulosebisphosphate carboxylase involves formation of a carbamate between an enzyme amino group and CO/sub 2/.

  9. Fructose-1,6-Bisphosphate Is an Allosteric Activator of Pyrophosphate:Fructose-6-Phosphate 1-Phosphotransferase.

    Science.gov (United States)

    Nielsen, T. H.

    1995-05-01

    The activity of highly purified pyrophosphate:fructose-6-phosphate 1-phosphotransferase (PFP) from barley (Hordeum vulgare) leaves was studied under conditions where the catalyzed reaction was allowed to approach equilibrium. The activity of PFP was monitored by determining the changes in the levels of fructose-6-phosphate, orthophosphate, and fructose-1,6-bisphosphate (Fru-1,6-bisP). Under these conditions PFP activity was not dependent on activation by fructose-2,6-bisphosphate (Fru-2,6-bisP). Inclusion of aldolase in the reaction mixture temporarily restored the dependence of PFP on Fru-2,6-bisP. Alternatively, PFP was activated by Fru-1,6-bisP in the presence of aldolase. It is concluded that Fru-1,6-bisP is an allosteric activator of barley PFP, which can substitute for Fru-2,6-bisP as an activator. A significant activation was observed at a concentration of 5 to 25 [mu]M Fru-1,6-bisP, which demonstrates that the allosteric site of barley PFP has a very high affinity for Fru-1,6-bisP. The high affinity for Fru-1,6-bisP at the allosteric site suggests that the observed activation of PFP by Fru-1,6-bisP constitutes a previously unrecognized in vivo regulation mechanism. PMID:12228454

  10. Insights into the carboxyltransferase reaction of pyruvate carboxylase from the structures of bound product and intermediate analogues

    OpenAIRE

    Lietzan, Adam D.; St. Maurice, Martin

    2013-01-01

    Pyruvate carboxylase (PC) is a biotin-dependent enzyme that catalyzes the MgATP- and bicarbonate-dependent carboxylation of pyruvate to oxaloacetate, an important anaplerotic reaction in central metabolism. The carboxyltransferase (CT) domain of PC catalyzes the transfer of a carboxyl group from carboxybiotin to the accepting substrate, pyruvate. It has been hypothesized that the reactive enolpyruvate intermediate is stabilized through a bidentate interaction with the metal ion in the CT doma...

  11. Biotin Uptake into Human Peripheral Blood Mononuclear Cells Increases Early in the Cell Cycle, Increasing Carboxylase Activities1,2

    OpenAIRE

    Stanley, J. Steven; Mock, Donald M.; Griffin, Jacob B.; Zempleni, Janos

    2002-01-01

    Cells respond to proliferation with increased accumulation of biotin, suggesting that proliferation enhances biotin demand. Here we determined whether peripheral blood mononuclear cells (PBMC) increase biotin uptake at specific phases of the cell cycle, and whether biotin is utilized to increase biotinylation of carboxylases. Biotin uptake was quantified in human PBMC that were arrested chemically at specific phases of the cell cycle, i.e., biotin uptake increased in the G1 phase of the cycle...

  12. 3-methylcrotonyl-CoA carboxylase deficiency: Clinical, biochemical, enzymatic and molecular studies in 88 individuals

    Directory of Open Access Journals (Sweden)

    Grünert Sarah C

    2012-05-01

    Full Text Available Abstract Background Isolated 3-methylcrotonyl-CoA carboxylase (MCC deficiency is an autosomal recessive disorder of leucine metabolism caused by mutations in MCCC1 or MCCC2 encoding the α and β subunit of MCC, respectively. The phenotype is highly variable ranging from acute neonatal onset with fatal outcome to asymptomatic adults. Methods We report clinical, biochemical, enzymatic and mutation data of 88 MCC deficient individuals, 53 identified by newborn screening, 26 diagnosed due to clinical symptoms or positive family history and 9 mothers, identified following the positive newborn screening result of their baby. Results Fifty-seven percent of patients were asymptomatic while 43% showed clinical symptoms, many of which were probably not related to MCC deficiency but due to ascertainment bias. However, 12 patients (5 of 53 identified by newborn screening presented with acute metabolic decompensations. We identified 15 novel MCCC1 and 16 novel MCCC2 mutant alleles. Additionally, we report expression studies on 3 MCCC1 and 8 MCCC2 mutations and show an overview of all 132 MCCC1 and MCCC2 variants known to date. Conclusions Our data confirm that MCC deficiency, despite low penetrance, may lead to a severe clinical phenotype resembling classical organic acidurias. However, neither the genotype nor the biochemical phenotype is helpful in predicting the clinical course.

  13. Relationship between NH4+ assimilation rate and in vivo phosphoenolpyruvate carboxylase activity

    International Nuclear Information System (INIS)

    The rate of NH4+ assimilation by N-limited Selenastrum minutum (Naeg.) Collins cells in the dark was set as an independent variable and the relationship between NH4+ assimilation rate and in vivo activity of phosphoenolpyruvate carboxylase (PEPC) was determined. In vivo activity of PEPC was measured by following the incorporation of H14CO3- into acid stable products. A linear relationship of 0.3 moles C fixed via PEPC per mole N assimilated was observed. This value agrees extremely well with the PEPC requirement for the synthesis of the amino acids found in total cellular protein. Determinations of metabolite levels in vivo at different rates of N assimilation indicated that the known metabolite effectors of S. minutum PEPC in vitro (KA Schuller, WC Plaxton, DH Turpin, [1990] Plant Physiol 93: 1303-1311) are important regulators of this enzyme during N assimilation. As PEPC activity increased in response to increasing rates of N assimilation, there was a corresponding decline in the level of PEPC inhibitors (2-oxoglutarate, malate), an increase in the level of PEPC activators (glutamine, dihydroxyacetone phosphate), and an increase in the Gln/Glu ratio. Treatment of N-limited cells with azaserine caused an increase in the Gln/Glu ratio resulting in increased PEPC activity in the absence of N assimilation. We suggest glutamate and glutamine play a key role in regulating the anaplerotic function of PEPC in this C3 organism

  14. Improving polyketide and fatty acid synthesis by engineering of the yeast acetyl-CoA carboxylase.

    Science.gov (United States)

    Choi, Jin Wook; Da Silva, Nancy A

    2014-10-10

    Polyketides and fatty acids are important in the production of pharmaceuticals, industrial chemicals, and biofuels. The synthesis of the malonyl-CoA building block, catalyzed by acetyl-CoA carboxylase (Acc1), is considered a limiting step to achieving high titers of polyketides and fatty acids in Saccharomyces cerevisiae. Acc1 is deactivated by AMP-activated serine/threonine protein kinase (Snf1) when glucose is depleted. To prevent this deactivation, the enzyme was aligned with the Rattus norvegicus (rat) Acc1 to identify a critical amino acid (Ser-1157) for phosphorylation and deactivation. Introduction of a S1157A mutation into Acc1 resulted in 9-fold higher specific activity following glucose depletion. The enzyme was tested in yeast engineered to produce the polyketide 6-methylsalisylic acid (6-MSA). Both 6-MSA and native fatty acid levels increased by 3-fold. Utilization of this modified Acc1 enzyme will also be beneficial for other products built from malonyl-CoA. PMID:25078432

  15. Cloning and characterization of cotton heteromeric acetyl-CoA carboxylase genes

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Heteromeric acetyl-coanzyme A(CoA)carboxylese(ACCase)catalyzes the formation of malonyl-CoA from acetyl-CoA.It plays an essential role in fatty acid synthesis in prokaryotes and most of plants.The heteromeric ACCase is composed of four subunits:biotin carboxylase (BC),biotin carboxyl carrier protein (BCCP),and α-and β-subunits of carboxyltransferese(α-andβ-CT).In this study,we cloned five novel genes encoding the subunits of heteromeric ACCese(one BC,BCCP and β-CT,and two α-CTs) from cotton (Gossypium hirsutum cv.zhongmian 35)by RACE-PCR.The deduced amino acid sequence of these cDNAs shares high similarity with other reported heteromeric ACCese subunits.The phylogenetic analysis indicated that the different subunits of heteromeric ACCase were grouped in a similar pattern.Southern blot analysis revealed the milti-copy patterns of these heteromeric ACCase genes in cotton genome.Semi-quantitative RT-PCR demonstrated that heteromeric ACCese genes were constitutively expressed in all of the cotton tissues,but the transcripts accumulated at a relatively low level in roots.To our knowledge,this is the first report about characterization of the heteromeric ACCase genes in cotton.

  16. Deregulation of feedback inhibition of phosphoenolpyruvate carboxylase for improved lysine production in Corynebacterium glutamicum.

    Science.gov (United States)

    Chen, Zhen; Bommareddy, Rajesh Reddy; Frank, Doinita; Rappert, Sugima; Zeng, An-Ping

    2014-02-01

    Allosteric regulation of phosphoenolpyruvate carboxylase (PEPC) controls the metabolic flux distribution of anaplerotic pathways. In this study, the feedback inhibition of Corynebacterium glutamicum PEPC was rationally deregulated, and its effect on metabolic flux redistribution was evaluated. Based on rational protein design, six PEPC mutants were designed, and all of them showed significantly reduced sensitivity toward aspartate and malate inhibition. Introducing one of the point mutations (N917G) into the ppc gene, encoding PEPC of the lysine-producing strain C. glutamicum LC298, resulted in ∼37% improved lysine production. In vitro enzyme assays and (13)C-based metabolic flux analysis showed ca. 20 and 30% increases in the PEPC activity and corresponding flux, respectively, in the mutant strain. Higher demand for NADPH in the mutant strain increased the flux toward pentose phosphate pathway, which increased the supply of NADPH for enhanced lysine production. The present study highlights the importance of allosteric regulation on the flux control of central metabolism. The strategy described here can also be implemented to improve other oxaloacetate-derived products. PMID:24334667

  17. Pyruvate carboxylase as a sensitive protein biomarker for exogenous steroid chemicals

    International Nuclear Information System (INIS)

    Assessing protein responses to endocrine disrupting chemicals is critical for understanding the mechanisms of chemical action and for the assessment of hazards. In this study, the response of the liver proteome of male rare minnows (Gobiocypris rarus) treated with 17β-estradiol (E2) and females treated with 17α-methyltestosterone (MT) were analyzed. A total of 23 and 24 proteins were identified with differential expression in response to E2 and MT, respectively. Pyruvate carboxylase (PC) was the only common differentially expressed protein in both males and females after E2- and MT-treatments. The mRNA as well as the protein levels of PC were significantly down-regulated compared with that of the controls (p < 0.05). Our results suggest that endocrine disruptors interfere with genes and proteins of the TCA cycle and PC may be a sensitive biomarker of exposure to exogenous steroid chemicals in the liver of fish. - Highlights: • The hepatic proteomes of rare minnow (Gobiocypris rarus) exposed to E2 and MT were analyzed. • Differentially expressed proteins (23 and 24 respectively) were identified following E2 and MT exposure. • Four differentially expressed proteins associated with chemical stimulus were characterized. • PC was identified as a responsive biomarker for both estrogens and androgens. - Our results suggest PC may be a sensitive biomarker of exposure to exogenous steroid chemicals in the liver of fish

  18. Multiple inter-kingdom horizontal gene transfers in the evolution of the phosphoenolpyruvate carboxylase gene family.

    Science.gov (United States)

    Peng, Yingmei; Cai, Jing; Wang, Wen; Su, Bing

    2012-01-01

    Pepcase is a gene encoding phosphoenolpyruvate carboxylase that exists in bacteria, archaea and plants,playing an important role in plant metabolism and development. Most plants have two or more pepcase genes belonging to two gene sub-families, while only one gene exists in other organisms. Previous research categorized one plant pepcase gene as plant-type pepcase (PTPC) while the other as bacteria-type pepcase (BTPC) because of its similarity with the pepcase gene found in bacteria. Phylogenetic reconstruction showed that PTPC is the ancestral lineage of plant pepcase, and that all bacteria, protistpepcase and BTPC in plants are derived from a lineage of pepcase closely related with PTPC in algae. However, their phylogeny contradicts the species tree and traditional chronology of organism evolution. Because the diversification of bacteria occurred much earlier than the origin of plants, presumably all bacterialpepcase derived from the ancestral PTPC of algal plants after divergingfrom the ancestor of vascular plant PTPC. To solve this contradiction, we reconstructed the phylogeny of pepcase gene family. Our result showed that both PTPC and BTPC are derived from an ancestral lineage of gamma-proteobacteriapepcases, possibly via an ancient inter-kingdom horizontal gene transfer (HGT) from bacteria to the eukaryotic common ancestor of plants, protists and cellular slime mold. Our phylogenetic analysis also found 48other pepcase genes originated from inter-kingdom HGTs. These results imply that inter-kingdom HGTs played important roles in the evolution of the pepcase gene family and furthermore that HGTsare a more frequent evolutionary event than previouslythought. PMID:23251445

  19. Multiple inter-kingdom horizontal gene transfers in the evolution of the phosphoenolpyruvate carboxylase gene family.

    Directory of Open Access Journals (Sweden)

    Yingmei Peng

    Full Text Available Pepcase is a gene encoding phosphoenolpyruvate carboxylase that exists in bacteria, archaea and plants,playing an important role in plant metabolism and development. Most plants have two or more pepcase genes belonging to two gene sub-families, while only one gene exists in other organisms. Previous research categorized one plant pepcase gene as plant-type pepcase (PTPC while the other as bacteria-type pepcase (BTPC because of its similarity with the pepcase gene found in bacteria. Phylogenetic reconstruction showed that PTPC is the ancestral lineage of plant pepcase, and that all bacteria, protistpepcase and BTPC in plants are derived from a lineage of pepcase closely related with PTPC in algae. However, their phylogeny contradicts the species tree and traditional chronology of organism evolution. Because the diversification of bacteria occurred much earlier than the origin of plants, presumably all bacterialpepcase derived from the ancestral PTPC of algal plants after divergingfrom the ancestor of vascular plant PTPC. To solve this contradiction, we reconstructed the phylogeny of pepcase gene family. Our result showed that both PTPC and BTPC are derived from an ancestral lineage of gamma-proteobacteriapepcases, possibly via an ancient inter-kingdom horizontal gene transfer (HGT from bacteria to the eukaryotic common ancestor of plants, protists and cellular slime mold. Our phylogenetic analysis also found 48other pepcase genes originated from inter-kingdom HGTs. These results imply that inter-kingdom HGTs played important roles in the evolution of the pepcase gene family and furthermore that HGTsare a more frequent evolutionary event than previouslythought.

  20. Characterization of the distal promoter of the human pyruvate carboxylase gene in pancreatic beta cells.

    Directory of Open Access Journals (Sweden)

    Ansaya Thonpho

    Full Text Available Pyruvate carboxylase (PC is an enzyme that plays a crucial role in many biosynthetic pathways in various tissues including glucose-stimulated insulin secretion. In the present study, we identify promoter usage of the human PC gene in pancreatic beta cells. The data show that in the human, two alternative promoters, proximal and distal, are responsible for the production of multiple mRNA isoforms as in the rat and mouse. RT-PCR analysis performed with cDNA prepared from human liver and islets showed that the distal promoter, but not the proximal promoter, of the human PC gene is active in pancreatic beta cells. A 1108 bp fragment of the human PC distal promoter was cloned and analyzed. It contains no TATA box but possesses two CCAAT boxes, and other putative transcription factor binding sites, similar to those of the distal promoter of rat PC gene. To localize the positive regulatory region in the human PC distal promoter, 5'-truncated and the 25-bp and 15-bp internal deletion mutants of the human PC distal promoter were generated and used in transient transfections in INS-1 832/13 insulinoma and HEK293T (kidney cell lines. The results indicated that positions -340 to -315 of the human PC distal promoter serve as (an activator element(s for cell-specific transcription factor, while the CCAAT box at -71/-67, a binding site for nuclear factor Y (NF-Y, as well as a GC box at -54/-39 of the human PC distal promoter act as activator sequences for basal transcription.

  1. Photosynthetic Characteristics and Heterosis in Transgenic Hybrid Rice with Maize Phosphoenolpyruvate Carboxylase (pepc) Gene

    Institute of Scientific and Technical Information of China (English)

    LI Ji-hang; XIANG Xun-chao; ZHOU Hua-qiang; HE Li-bin; ZHANG Kai-zheng; LI Ping

    2006-01-01

    Three F1 hybrids derived from the sterile rice lines Gang 46A, 776A and 2480A and the improved restorer line Shuhui 881 containing maize phosphoenolpyruvate carboxylase (pepc) gene were used to analyze the effect of pepc gene on the heterosis and photosynthetic characteristics, while the F1 obtained by crossing Shuhui 881 with the above three sterile lines served as controls. The dynamics of photosynthetic characteristics in leaves of three F1 with pepc gene and their controls were determined at the initial-tillering, maxium-tillering, elongation, initial-heading, heading, maturity stages, and other different times after flag leaf fully expanded. The PEPCase activities of the three F1 with pepc gene increased significantly as compared with control plants during the whole developmental stages. Moreover, the net photosynthesis rate (Pn) also increased to certain extent. The data showed that PEPCase activity was significantly correlated to Pn with a correlation coefficient of 0.6081**. The photosynthetic indexes of the three F1 with pepc gene were obviously superior with respective to controls in apparent quantum yield, light compensation point (LCP) and carboxylation efficiency (CE), while the CO2 compensation point (CCP) was lower than that of corresponding control. The Pn of the three F1 with pepc gene at light saturation point (LSP) and CO2 saturation point (CSP) was also higher than that of control plants. In addition, the three F1 with pepc gene had an average increase of 37.10% in grain yields per plant in comparison with control plants. The results indicated that the photosynthetic characteristics of hybrid rice containing pepc gene had been improved to some extent due to the introduction of pepc gene.

  2. Effects of phosphoenolpyruvate carboxylase desensitization on glutamic acid production in Corynebacterium glutamicum ATCC 13032.

    Science.gov (United States)

    Wada, Masaru; Sawada, Kazunori; Ogura, Kotaro; Shimono, Yuta; Hagiwara, Takuya; Sugimoto, Masakazu; Onuki, Akiko; Yokota, Atsushi

    2016-02-01

    Phosphoenolpyruvate carboxylase (PEPC) in Corynebacterium glutamicum ATCC13032, a glutamic-acid producing actinobacterium, is subject to feedback inhibition by metabolic intermediates such as aspartic acid and 2-oxoglutaric acid, which implies the importance of PEPC in replenishing oxaloacetic acid into the TCA cycle. Here, we investigated the effects of feedback-insensitive PEPC on glutamic acid production. A single amino-acid substitution in PEPC, D299N, was found to relieve the feedback control by aspartic acid, but not by 2-oxoglutaric acid. A simple mutant, strain R1, having the D299N substitution in PEPC was constructed from ATCC 13032 using the double-crossover chromosome replacement technique. Strain R1 produced glutamic acid at a concentration of 31.0 g/L from 100 g/L glucose in a jar fermentor culture under biotin-limited conditions, which was significantly higher than that of the parent, 26.0 g/L (1.19-fold), indicative of the positive effect of desensitized PEPC on glutamic acid production. Another mutant, strain DR1, having both desensitized PEPC and PYK-gene deleted mutations, was constructed in a similar manner using strain D1 with a PYK-gene deleted mutation as the parent. This mutation had been shown to enhance glutamic acid production in our previous study. Although marginal, strain D1 produced higher glutamic acid, 28.8 g/L, than ATCC13032 (1.11-fold). In contrast, glutamic acid production by strain DR-1 was elevated up to 36.9 g/L, which was 1.42-fold higher than ATCC13032 and significantly higher than the other three strains. The results showed a synergistic effect of these two mutations on glutamic acid production in C. glutamicum. PMID:26168906

  3. Epigenetic regulation of pyruvate carboxylase gene expression in the postpartum liver.

    Science.gov (United States)

    Walker, C G; Crookenden, M A; Henty, K M; Handley, R R; Kuhn-Sherlock, B; White, H M; Donkin, S S; Snell, R G; Meier, S; Heiser, A; Loor, J J; Mitchell, M D; Roche, J R

    2016-07-01

    Hepatic gluconeogenesis is essential for maintenance of whole body glucose homeostasis and glucose supply for mammary lactose synthesis in the dairy cow. Upregulation of the gluconeogenic enzyme pyruvate carboxylase (PC) during the transition period is vital in the adaptation to the greater glucose demands associated with peripartum lactogenesis. The objective of this study was to determine if PC transcription in hepatocytes is regulated by DNA methylation and if treatment with a nonsteroidal anti-inflammatory drug (NSAID) alters methylation of an upstream DNA sequence defined as promoter 1. Dairy cows were left untreated (n=20), or treated with a NSAID during the first 5 d postcalving (n=20). Liver was biopsied at d 7 precalving and d 7, 14, and 28 postcalving. Total PC and transcript specific gene expression was quantified using quantitative PCR and DNA methylation of promoter 1 was quantified using bisulfite Sanger sequencing. Expression of PC changed over the transition period, with increased expression postcalving occurring concurrently with increased circulating concentration of nonesterified fatty acids. The DNA methylation percentage was variable at all sites quantified and ranged from 21 to 54% across the 15 CpG dinucleotides within promoter 1. The DNA methylation at wk 1 postcalving, however, was not correlated with gene expression of promoter 1-regulated transcripts and we did not detect an effect of NSAID treatment on DNA methylation or PC gene expression. Our results do not support a role for DNA methylation in regulating promoter 1-driven gene expression of PC at wk 1 postcalving. Further research is required to determine the mechanisms regulating increased PC expression over the transition period. PMID:27085418

  4. Active Site Loop Dynamics of a Class IIa Fructose 1,6-Bisphosphate Aldolase from Mycobacterium tuberculosis

    Energy Technology Data Exchange (ETDEWEB)

    Pegan, Scott D. [Univ. of Denver, CO (United States); Rukseree, Kamolchanok [National Center for Genetic Engineering and Biotechnology (BIOTEC), Tha Khlong (Thailand); Capodagli, Glenn C. [Univ. of Denver, CO (United States); Baker, Erica A. [Univ. of Denver, CO (United States); Krasnykh, Olga [Univ. of Illinois, Chicago, IL (United States); Franzblau, Scott G. [Univ. of Illinois, Chicago, IL (United States); Mesecar, Andrew D. [Purdue Univ., West Lafayette, IN (United States)

    2013-01-08

    The class II fructose 1,6-bisphosphate aldolases (FBAs, EC 4.1.2.13) comprises one of two families of aldolases. Instead of forming a Schiff base intermediate using an ε-amino group of a lysine side chain, class II FBAs utilize Zn(II) to stabilize a proposed hydroxyenolate intermediate (HEI) in the reversible cleavage of fructose 1,6-bisphosphate, forming glyceraldehyde 3-phosphate and dihydroxyacetone phosphate (DHAP). As class II FBAs have been shown to be essential in pathogenic bacteria, focus has been placed on these enzymes as potential antibacterial targets. Although structural studies of class II FBAs from Mycobacterium tuberculosis (MtFBA), other bacteria, and protozoa have been reported, the structure of the active site loop responsible for catalyzing the protonation–deprotonation steps of the reaction for class II FBAs has not yet been observed. We therefore utilized the potent class II FBA inhibitor phosphoglycolohydroxamate (PGH) as a mimic of the HEI- and DHAP-bound form of the enzyme and determined the X-ray structure of the MtFBA–PGH complex to 1.58 Å. Remarkably, we are able to observe well-defined electron density for the previously elusive active site loop of MtFBA trapped in a catalytically competent orientation. Utilization of this structural information and site-directed mutagenesis and kinetic studies conducted on a series of residues within the active site loop revealed that E169 facilitates a water-mediated deprotonation–protonation step of the MtFBA reaction mechanism. Furthermore, solvent isotope effects on MtFBA and catalytically relevant mutants were used to probe the effect of loop flexibility on catalytic efficiency. Additionally, we also reveal the structure of MtFBA in its holoenzyme form.

  5. Low mobility of phosphatidylinositol 4,5-bisphosphate underlies receptor specificity of Gq-mediated ion channel regulation in atrial myocytes

    OpenAIRE

    Cho, Hana; Kim, Yeon A; Yoon, Jin-Young; Lee, Doyun; Kim, Jae Ho; Lee, Suk Ho; Ho, Won-Kyung

    2005-01-01

    We have shown previously that cardiac G protein-gated inwardly rectifying K+ (GIRK) channels are inhibited by Gq protein-coupled receptors (GqPCRs) via phosphatidylinositol 4,5-bisphosphate (PIP2) depletion in a receptor-specific manner. To investigate the mechanism of receptor specificity, we examined whether the activation of GqPCRs induces localized PIP2 depletion. When we applied endothelin-1 to the bath, GIRK channel activities recorded in cell-attached patches were not changed, implying...

  6. Ion-exchange chromatography separates activities synthesizing and degrading fructose 2,6-bisphosphate from C3 and C4 leaves but not from rat liver

    Science.gov (United States)

    Macdonald, F. D.; Chou, Q.; Buchanan, B. B.

    1987-01-01

    Fructose-6-phosphate,2-kinase and fructose-2,6-bisphosphatase were separated on the basis of charge from leaves of C3 (spinach, lettuce, and pea) and C4 (sorghum and amaranthus) plants but not from rat liver--a tissue known to contain a bifunctional enzyme with both activities. [2-32P]Fructose 2,6-bisphosphate binding experiments also suggest that the major forms of these activities reside on different proteins in leaves.

  7. Main: 1WDD [RPSD[Archive

    Lifescience Database Archive (English)

    Full Text Available 1WDD イネ Rice Oryza sativa L. Ribulose Bisphosphate Carboxylase Large Chain Precursor Name=Rbcl ... binitol-1, 5-Bisphosphate To Be Published Rubisco, Photosynthesis , Alpha/Beta Barrel, N-Methylmethionine, Post-Trans ...

  8. A preliminary X-ray study of d,d-heptose-1,7-bisphosphate phosphatase from Burkholderia thailandensis E264

    International Nuclear Information System (INIS)

    In this study, d,d-heptose-1,7-bisphosphate phosphatase has been cloned, expressed, purified and crystallized. d,d-Heptose-1,7-bisphosphate phosphatase (GmhB), which is involved in the third step of the NDP-heptose biosynthesis pathway, converts d,d-heptose-1,7-bisphosphate to d,d-heptose-1-phosphate. This biosynthesis pathway is a target for new antibiotics or antibiotic adjuvants for Gram-negative pathogens. Burkholderia thailandensis is a useful surrogate organism for studying the pathogenicity of melioidosis owing to its extensive genomic similarity to B. pseudomallei. Melioidosis caused by B. pseudomallei is a serious invasive disease of animals and humans in tropical and subtropical areas. In this study, GmhB has been cloned, expressed, purified and crystallized. X-ray data have also been collected to 2.50 Å resolution using synchrotron radiation. The crystal belonged to space group P6, with unit-cell parameters a = 243.2, b = 243.2, c = 41.1 Å

  9. Co-expression of Triosephosphate Isomerase, Fructose-1, 6-bisphosphate Aldolase and Fructose-1, 6-bisphosphatase in E.coli.

    Science.gov (United States)

    Tang, Gong-Li; Yang, Chun-Song; Bao, Jian-Shao; Wang, Yan-Fang; Chen, Hai-Bao; Shi, Ding-Ji; Liu, Feng-Long

    2001-01-01

    To establish a way to control or to decrease the daily increasing concentration of atmospheric CO(2), metabolically engineering Cyanobacteria was taken for the improvement of its efficiency of photosynthetic CO(2) fixation. As a preliminary stage of this study, three genes coding for three important Calvin cycle enzymes, i.e. triosephosphate isomerase (TPI), fructose-1, 6-bisphosphate aldolase(FBP aldolase),and fructose-1, 6-bisphosphatase(FBPase), respectively, have been cloned into one plasmid, pTrcFAT, which is controlled by promoter trc. Successful co-transcriptional expression of these three genes resulted inhigh yields of these enzymes under the induction of 0.25 mmol/L IPTG. Bioassay showed that the expressed enzymes from one liter of culture could directly catalyze DHAP conversion into 700 &mgr;mol of fructose-6-phosphate (F-6-P) per one minute. Furthermore, in order to introduce the three genes co-expression system into Cyanobacteria, a shuttle plasmid between E.coli and Cyanobacteria was constructed using plasmid pTrcFAT and a shuttle vector pDC-8, forming ashuttle plasmid pDCFAT-2 containing a dimer of the three genes co-expression operator. Successful co-expression in E.coli of pDCFAT-2 with higher full activity has been obtained. This shuttle was used to transform of Cyanobacteria Synechococcus sp. PCC 7942, and a few positive colonies were obtained. PMID:12053203

  10. A conformational investigation of propeptide binding to the integral membrane protein γ-glutamyl carboxylase using nanodisc hydrogen exchange mass spectrometry

    DEFF Research Database (Denmark)

    Parker, Christine H; Morgan, Christopher R; Rand, Kasper Dyrberg;

    2014-01-01

    Gamma (γ)-glutamyl carboxylase (GGCX) is an integral membrane protein responsible for the post-translational catalytic conversion of select glutamic acid (Glu) residues to γ-carboxy glutamic acid (Gla) in vitamin K-dependent (VKD) proteins. Understanding the mechanism of carboxylation and the rol...

  11. Serine-15 is the regulatory seryl-phosphorylation site in C4-leaf phosphoenolpyruvate carboxylase (PEPCase) from maize

    International Nuclear Information System (INIS)

    The 32P-labeled regulatory site phosphopeptide was purified from a tryptic digest of in vitro phosphorylated/activated dark-form PEPCase by metal ion affinity and reversed-phase chromatography and subjected to automated Edman degradation analysis. The amino acid sequence of this phosphoseryl peptide is His-His-Ser(P)-Ile-Asp-Ala-Gln-Leu-Arg. This nonapeptide, which corresponds exactly to residues 13-21 in the deduced primary sequence of the maize leaf carboxylase, is far removed from a recently identified active-site cysteine (Cys-553) in the C-terminal region of the primary structure. Comparative analysis of the deduced N-terminal sequences of C3, C4, and CAM leaf PEPCases suggests that the motif of Lys/Arg-X-X-Ser is an important structural requirement of the C4- and CAM-leaf protein-serine kinases

  12. Serine-15 is the regulatory seryl-phosphorylation site in C sub 4 -leaf phosphoenolpyruvate carboxylase (PEPCase) from maize

    Energy Technology Data Exchange (ETDEWEB)

    Jiao, Jinan; Chollet, R. (Univ. of Nebraska, Lincoln (USA))

    1990-05-01

    The {sup 32}P-labeled regulatory site phosphopeptide was purified from a tryptic digest of in vitro phosphorylated/activated dark-form PEPCase by metal ion affinity and reversed-phase chromatography and subjected to automated Edman degradation analysis. The amino acid sequence of this phosphoseryl peptide is His-His-Ser(P)-Ile-Asp-Ala-Gln-Leu-Arg. This nonapeptide, which corresponds exactly to residues 13-21 in the deduced primary sequence of the maize leaf carboxylase, is far removed from a recently identified active-site cysteine (Cys-553) in the C-terminal region of the primary structure. Comparative analysis of the deduced N-terminal sequences of C{sub 3}, C{sub 4}, and CAM leaf PEPCases suggests that the motif of Lys/Arg-X-X-Ser is an important structural requirement of the C{sub 4}- and CAM-leaf protein-serine kinases.

  13. Phosphorylation-dephosphorylation process as a probable mechanism for the diurnal regulatory changes of phosphoenolpyruvate carboxylase in CAM plants.

    Science.gov (United States)

    Brulfert, J; Vidal, J; Le Marechal, P; Gadal, P; Queiroz, O; Kluge, M; Kruger, I

    1986-04-14

    Day and night forms of phosphoenolpyruvate carboxylase (EC 4.1.1.31) (PEPC) were extracted from leaves of the CAM plants Kalanchoe daigremontiana, K. tubiflora and K. blossfeldiana previously fed with [32P] labelled phosphate solution. A one-step immunochemical purification followed by SDS polyacrylamide gel electrophoresis and autoradiography showed that, in all species, the night form of the enzyme was phosphorylated and not the day form. Limited acid hydrolysis of the night form and two-dimensional separation identified predominantly labelled phosphoserine and phosphothreonine. In vitro addition of exogenous acid phosphatase (EC 3.1.3.2) to desalted night form-containing extracts resulted within 30 min in a shift in PEPC enzymic properties similar to the in vivo changes from night to day form. It is suggested that phosphorylation-dephosphorylation of the enzyme could be the primary in vivo process which might explain the observed rhythmicity of enzymic properties. PMID:3707571

  14. Determination of ploidy level and isolation of genes encoding acetyl-CoA carboxylase in Japanese Foxtail (Alopecurus japonicus.

    Directory of Open Access Journals (Sweden)

    Hongle Xu

    Full Text Available Ploidy level is important in biodiversity studies and in developing strategies for isolating important plant genes. Many herbicide-resistant weed species are polyploids, but our understanding of these polyploid weeds is limited. Japanese foxtail, a noxious agricultural grass weed, has evolved herbicide resistance. However, most studies on this weed have ignored the fact that there are multiple copies of target genes. This may complicate the study of resistance mechanisms. Japanese foxtail was found to be a tetraploid by flow cytometer and chromosome counting, two commonly used methods in the determination of ploidy levels. We found that there are two copies of the gene encoding plastidic acetyl-CoA carboxylase (ACCase in Japanese foxtail and all the homologous genes are expressed. Additionally, no difference in ploidy levels or ACCase gene copy numbers was observed between an ACCase-inhibiting herbicide-resistant and a herbicide-sensitive population in this study.

  15. Cloning, Expression and Purification of Wheat Acetyl-CoA Carboxylases CT Domain in E.coil

    Institute of Scientific and Technical Information of China (English)

    WANG Rui-jian; YANG Xue-ying; ZHENG Liang-yu; YANG Ye; GAO Gui; CAO Shu-gui

    2008-01-01

    The entire gene of carboxyltransferase(CT) domain of acetyl-CoA carboxylase(ACCase) from Chinese Spring wheat(CSW) plastid was cloned firstly,and the 2.3 kb gene was inserted into PET28a+ vector and expressed in E.coil in a soluble state.The (His)6 fusion protein was identified by SDS-PAGE and Western blot.The recombinant protein was purified by affinity chromatography,and the calculated molecular mass(Mr) was 88000.The results of the sequence analysis indicate that the cloned gene(GeneBank accession No.EU124675) was a supplement and revision of the reported ACCase CT partial cDNA from Chinese Spring wheat plastid.The recombinant protein will be significant for us to investigate the recognizing mechanism between ACCase and herbicides,and further to screen new herbicides.

  16. A 3-methylcrotonyl-CoA carboxylase deficient human skin fibroblast transcriptome reveals underlying mitochondrial dysfunction and oxidative stress.

    Science.gov (United States)

    Zandberg, L; van Dyk, H C; van der Westhuizen, F H; van Dijk, A A

    2016-09-01

    Isolated 3-methylcrotonyl-CoA carboxylase (MCC) deficiency is an autosomal recessive inherited metabolic disease of leucine catabolism with a highly variable phenotype. Apart from extensive mutation analyses of the MCCC1 and MCCC2 genes encoding 3-methylcrotonyl-CoA carboxylase (EC 6.4.1.4), molecular data on MCC deficiency gene expression studies in human tissues is lacking. For IEMs, unbiased '-omics' approaches are starting to reveal the secondary cellular responses to defects in biochemical pathways. Here we present the first whole genome expression profile of immortalized cultured skin fibroblast cells of two clinically affected MCC deficient patients and two healthy individuals generated using Affymetrix(®)HuExST1.0 arrays. There were 16191 significantly differentially expressed transcript IDs of which 3591 were well annotated and present in the predefined knowledge database of Ingenuity Pathway Analysis software used for downstream functional analyses. The most noticeable feature of this MCCA deficient skin fibroblast transcriptome was the typical genetic hallmark of mitochondrial dysfunction, decreased antioxidant response and disruption of energy homeostasis, which was confirmed by mitochondrial functional analyses. The MCC deficient transcriptome seems to predict oxidative stress that could alter the complex secondary cellular response that involve genes of the glycolysis, the TCA cycle, OXPHOS, gluconeogenesis, β-oxidation and the branched-chain fatty acid metabolism. An important emerging insight from this human MCCA transcriptome in combination with previous reports is that chronic exposure to the primary and secondary metabolites of MCC deficiency and the resulting oxidative stress might impact adversely on the quality of life and energy levels, irrespective of whether MCC deficient individuals are clinically affected or asymptomatic. PMID:27417235

  17. AcEST: DK960177 [AcEST

    Lifescience Database Archive (English)

    Full Text Available 9. 5' end sequence. DK960177 CL2Contig2 Show DK960177 Clone id TST39A01NGRL0006_L09 Library TST39 Length 739... Definition Adiantum capillus-veneris mRNA. clone: TST39A01NGRL0006_L09. 5' end sequence. Accession DK960177...tabase search programs, Nucleic Acids Res. 25:3389-3402. Query= DK960177|Adiantum capillus-veneris mRNA, clo...ylase small ch... 178 3e-44 sp|P16031|RBS_LARLA Ribulose bisphosphate carboxylase small chai... 177 4e-44 sp...|Q40004|RBS_HORVU Ribulose bisphosphate carboxylase small chai... 177 4e-44 sp|P1

  18. Inactive enzymatic mutant proteins (phosphoglycerate mutase and enolase as sugar binders for ribulose-1,5-bisphosphate regeneration reactors

    Directory of Open Access Journals (Sweden)

    Giri Ashok

    2005-02-01

    Full Text Available Abstract Background Carbon dioxide fixation bioprocess in reactors necessitates recycling of D-ribulose1,5-bisphosphate (RuBP for continuous operation. A radically new close loop of RuBP regenerating reactor design has been proposed that will harbor enzyme-complexes instead of purified enzymes. These reactors will need binders enabling selective capture and release of sugar and intermediate metabolites enabling specific conversions during regeneration. In the current manuscript we describe properties of proteins that will act as potential binders in RuBP regeneration reactors. Results We demonstrate specific binding of 3-phosphoglycerate (3PGA and 3-phosphoglyceraldehyde (3PGAL from sugar mixtures by inactive mutant of yeast enzymes phosphoglycerate mutase and enolase. The reversibility in binding with respect to pH and EDTA has also been shown. No chemical conversion of incubated sugars or sugar intermediate metabolites were found by the inactive enzymatic proteins. The dissociation constants for sugar metabolites are in the micromolar range, both proteins showed lower dissociation constant (Kd for 3-phosphoglycerate (655–796 μM compared to 3-phosphoglyceraldehyde (822–966 μM indicating higher affinity for 3PGA. The proteins did not show binding to glucose, sucrose or fructose within the sensitivity limits of detection. Phosphoglycerate mutase showed slightly lower stability on repeated use than enolase mutants. Conclusions The sugar and their intermediate metabolite binders may have a useful role in RuBP regeneration reactors. The reversibility of binding with respect to changes in physicochemical factors and stability when subjected to repeated changes in these conditions are expected to make the mutant proteins candidates for in-situ removal of sugar intermediate metabolites for forward driving of specific reactions in enzyme-complex reactors.

  19. Plasma membrane associated phospholipase C from human platelets: Synergistic stimulation of phosphatidylinositol 4,5-bisphosphate hydrolysis by thrombin and guanosine 5'-O-(3-thiotriphosphate)

    International Nuclear Information System (INIS)

    The effects of thrombin and GTPγS on the hydrolysis of phosphoinositides by membrane-associated phospholipase C (PLC) from human platelets were examined with endogenous [3H]inositol-labeled membranes or with lipid vesicles containing either [3H]phosphatidylinositol or [3H]phosphatidylinositol 4,5-bisphosphate. GTPγS (1 μM) or thrombin (1 unit/mL) did not stimulate release of inositol trisphosphate (IP3), inositol bisphosphate (IP2), or inositol phosphate (IP) from [3H]inositol-labeled membranes. IP2 and IP3, but not IP, from [3H]inositol-labeled membranes were, however, stimulated 3-fold by GTPγS (1 μM) plus thrombin (1 unit/mL). A higher concentration of GTPγS (100 μM) alone also stimulated IP2 and IP3, but not IP, release. In the presence of 1 mM calcium, release of IP2 and IP3 was increased 6-fold over basal levels; however, formation of IP was not observed. At submicromolar calcium concentration, hydrolysis of exogenous phosphatidylinositol 4,5-bisphosphate (PIP2) by platelet membrane associated PLC was also markedly enhanced by GTPγS (100 μM) or GTPγS (1 μM) plus thrombin (1 unit/mL). Under identical conditions, exogenous phosphatidylinositol (PI) was not hydrolyzed. The same substrate specificity was observed when the membrane-associated PLC was activated with 1 mM calcium. Thrombin-induced hydrolysis of PIP2 was inhibited by treatment of the membranes with pertussis toxin or pretreatment of intact platelets with 12-O-tetradecanoyl-13-acetate (TPA) prior to preparation of membranes. Pertussis toxin did not inhibit GTPγS (100 μM) or calcium (1 mM) dependent PIP2 breakdown, while TPA inhibited GTPγS-dependent but not calcium-dependent phospholipase C activity

  20. Oxygen Requirement and Inhibition of C4 Photosynthesis . An Analysis of C4 Plants Deficient in the C3 and C4 Cycles

    OpenAIRE

    Maroco, J.P.; Ku, M.S.B.; Lea, P J; Dever, L.V.; Leegood, R C; Furbank, R.T.; Edwards, G. E.

    1998-01-01

    The basis for O2 sensitivity of C4 photosynthesis was evaluated using a C4-cycle-limited mutant of Amaranthus edulis (a phosphoenolpyruvate carboxylase-deficient mutant), and a C3-cycle-limited transformant of Flaveria bidentis (an antisense ribulose-1,5-bisphosphate carboxylase/oxygenase [Rubisco] small subunit transformant). Data obtained with the C4-cycle-limited mutant showed that atmospheric levels of O2 (20 kPa) caused increased inhibition of photosynthesis as a result of higher levels ...

  1. High yield synthesis of 14C labelled intermediates of the L-type pentose pathway: octulose mono- and biphosphates, sedoheptulose 1,7-bisphosphate and D-arabinose 5-phosphate

    International Nuclear Information System (INIS)

    Methods for the enzymic synthesis, isolation, purification and analysis of the 14C labelled intermediates that are characteristic of the L-type pentose phosphate pathway are described. These are D-glycero D-ido octulose 1,8-bisphosphate and 8-monophosphate; D-glycero D-altro octulose mono- and bisphosphates; sedoheptulose 1,7-bisphosphate and D-arabinose 5-phosphate. The authenticity of each of the 14C-labelled sugar phosphates was confirmed using a variety of chromatographic methods, enzymatic analytic methods and NMR spectroscopy. The 14C labelled compounds are used in investigations involving the search for the identity of the pentose pathway in tissues in vitro, for the measurement of L-type pathway enzyme reactions and for testing the mechanistic predictions of the L-type pathway in vitro. (author)

  2. AMPK activation represses the human gene promoter of the cardiac isoform of acetyl-CoA carboxylase: Role of nuclear respiratory factor-1

    International Nuclear Information System (INIS)

    Research highlights: → AMPK inhibits acetyl-CoA carboxylase beta gene promoter activity. → Nuclear respiratory factor-1 inhibits acetyl-CoA carboxylase beta promoter activity. → AMPK regulates acetyl-CoA carboxylase beta at transcriptional level. -- Abstract: The cardiac-enriched isoform of acetyl-CoA carboxylase (ACCβ) produces malonyl-CoA, a potent inhibitor of carnitine palmitoyltransferase-1. AMPK inhibits ACCβ activity, lowering malonyl-CoA levels and promoting mitochondrial fatty acid β-oxidation. Previously, AMPK increased promoter binding of nuclear respiratory factor-1 (NRF-1), a pivotal transcriptional modulator controlling gene expression of mitochondrial proteins. We therefore hypothesized that NRF-1 inhibits myocardial ACCβ promoter activity via AMPK activation. A human ACCβ promoter-luciferase construct was transiently transfected into neonatal cardiomyocytes ± a NRF-1 expression construct. NRF-1 overexpression decreased ACCβ gene promoter activity by 71 ± 4.6% (p < 0.001 vs. control). Transfections with 5'-end serial promoter deletions revealed that NRF-1-mediated repression of ACCβ was abolished with a pPIIβ-18/+65-Luc deletion construct. AMPK activation dose-dependently reduced ACCβ promoter activity, while NRF-1 addition did not further decrease it. We also investigated NRF-1 inhibition in the presence of upstream stimulatory factor 1 (USF1), a known transactivator of the human ACCβ gene promoter. Here NRF-1 blunted USF1-dependent induction of ACCβ promoter activity by 58 ± 7.5% (p < 0.001 vs. control), reversed with a dominant negative NRF-1 construct. NRF-1 also suppressed endogenous USF1 transcriptional activity by 55 ± 6.2% (p < 0.001 vs. control). This study demonstrates that NRF-1 is a novel transcriptional inhibitor of the human ACCβ gene promoter in the mammalian heart. Our data extends AMPK regulation of ACCβ to the transcriptional level.

  3. Measurement of Acylcarnitine Substrate to Product Ratios Specific to Biotin-Dependent Carboxylases Offers a Combination of Indicators of Biotin Status in Humans12

    OpenAIRE

    Bogusiewicz, Anna; Horvath, Thomas D; Stratton, Shawna L.; Mock, Donald M; Boysen, Gunnar

    2012-01-01

    This work describes a novel liquid chromatography tandem MS (LC-MS/MS) method for the determination of ratios of acylcarnitines arising from acyl-CoA substrates and products that reflect metabolic disturbances caused by marginal biotin deficiency. The urinary ratios reflecting reduced activities of biotin-dependent enzymes include the following: 1) the ratio of 3-hydroxyisovalerylcarnitine : 3-methylglutarylcarnitine (3HIAc : MGc) for methylcrotonyl-CoA carboxylase; 2) the ratio of propionylc...

  4. Light-stimulated transcription of genes for two chloroplast polypeptides in isolated pea leaf nuclei

    OpenAIRE

    Gallagher, Thomas F; Ellis, R. John

    1982-01-01

    Nuclei isolated from both light-grown and dark-grown leaves of Pisum sativum by Percoll density gradient centrifugation incorporate labelled UTP into RNA when supplemented with the other three nucleoside triphosphates. The RNA is heterodisperse, with transcripts up to at least 25S in size. Among these transcripts are sequences hybridizing to cloned DNA probes for wheat rRNA and two abundant chloroplast polypeptides of Pisum, viz. the small subunit of ribulose bisphosphate carboxylase and a po...

  5. Targeting of proteins to the thylakoid lumen by the bipartite transit peptide of the 33 kd oxygen-evolving protein.

    OpenAIRE

    Ko, K.; Cashmore, A. R.

    1989-01-01

    Various chimeric precursors and deletions of the 33 kd oxygen-evolving protein (OEE1) were constructed to study the mechanism by which chloroplast proteins are imported and targeted to the thylakoid lumen. The native OEE1 precursor was imported into isolated chloroplasts, processed and localized in the thylakoid lumen. Replacement of the OEE1 transit peptide with the transit peptide of the small subunit of ribulose-1,5-bisphosphate carboxylase, a stromal protein, resulted in redirection of ma...

  6. Acetyl-CoA carboxylase inhibition by ND-630 reduces hepatic steatosis, improves insulin sensitivity, and modulates dyslipidemia in rats.

    Science.gov (United States)

    Harriman, Geraldine; Greenwood, Jeremy; Bhat, Sathesh; Huang, Xinyi; Wang, Ruiying; Paul, Debamita; Tong, Liang; Saha, Asish K; Westlin, William F; Kapeller, Rosana; Harwood, H James

    2016-03-29

    Simultaneous inhibition of the acetyl-CoA carboxylase (ACC) isozymes ACC1 and ACC2 results in concomitant inhibition of fatty acid synthesis and stimulation of fatty acid oxidation and may favorably affect the morbidity and mortality associated with obesity, diabetes, and fatty liver disease. Using structure-based drug design, we have identified a series of potent allosteric protein-protein interaction inhibitors, exemplified by ND-630, that interact within the ACC phosphopeptide acceptor and dimerization site to prevent dimerization and inhibit the enzymatic activity of both ACC isozymes, reduce fatty acid synthesis and stimulate fatty acid oxidation in cultured cells and in animals, and exhibit favorable drug-like properties. When administered chronically to rats with diet-induced obesity, ND-630 reduces hepatic steatosis, improves insulin sensitivity, reduces weight gain without affecting food intake, and favorably affects dyslipidemia. When administered chronically to Zucker diabetic fatty rats, ND-630 reduces hepatic steatosis, improves glucose-stimulated insulin secretion, and reduces hemoglobin A1c (0.9% reduction). Together, these data suggest that ACC inhibition by representatives of this series may be useful in treating a variety of metabolic disorders, including metabolic syndrome, type 2 diabetes mellitus, and fatty liver disease. PMID:26976583

  7. Promotive Effect of Low Concentrations of NaHSO3 on Photophosphorylation and Photosynthesis in Phosphoenolpyruvate Carboxylase Transgenic Rice Leaves

    Institute of Scientific and Technical Information of China (English)

    Ben-Hua JI; Hong-He TAN; Rong ZHOU; De-Mao JIAO; Yun-Gang SHEN

    2005-01-01

    Spraying a 1-2 mmol/L solution of NaHSO3 on the leaves of wild-type rice (Oryza sativa L.)Kitaake (WT), phosphoenolpyruvate carboxylase (PEPC) transgenic (PC) rice and PEPC+phosphate dikinase (PPDK) transgenic rice (PC+PK), in which the germplasm was transformed with wild-type Kitaake as the gene receptor, resulted in an enhancement of the net photosynthetic rate by 23.0%, 28.8%, and 34.4%,respectively, for more than 3 d. It was also observed that NaHSO3 application caused an increase in the ATP content in leaves. Spraying PMS (a cofactor catalysing the photophosphorylation cycle) and NaHSO3 separately or together on leaves resulted in an increase in photosynthesis with all treatments. There was no additional effect on photosynthetic rate when the mixture was applied, suggesting that the mechanism by which NaHSO3 promotes photosynthesis is similar to the mechanism by which PMS acts and that both of compounds enhanced the supply of ATP. After spraying a solution of NaHSO3 on leaves, compared with the WT Kitaake rice, a greater enhancement of net photosynthetic rate was observed in PEPC transgenic (PC) and PEPC+PPDK transgenic (PC+PK) rice, with the greatest increase being observed in the latter group. Therefore ATP supply may become the limiting factor that concentrates CO2 in rice leaves transformed with an exogenous PEPC gene and exogenous PEPC+PPDK genes.

  8. Osteoblast-Specific γ-Glutamyl Carboxylase-Deficient Mice Display Enhanced Bone Formation With Aberrant Mineralization.

    Science.gov (United States)

    Azuma, Kotaro; Shiba, Sachiko; Hasegawa, Tomoka; Ikeda, Kazuhiro; Urano, Tomohiko; Horie-Inoue, Kuniko; Ouchi, Yasuyoshi; Amizuka, Norio; Inoue, Satoshi

    2015-07-01

    Vitamin K is a fat-soluble vitamin that is necessary for blood coagulation. In addition, it has bone-protective effects. Vitamin K functions as a cofactor of γ-glutamyl carboxylase (GGCX), which activates its substrates by carboxylation. These substrates are found throughout the body and examples include hepatic blood coagulation factors. Furthermore, vitamin K functions as a ligand of the nuclear receptor known as steroid and xenobiotic receptor (SXR) and its murine ortholog, pregnane X receptor (PXR). We have previously reported on the bone-protective role of SXR/PXR signaling by demonstrating that systemic Pxr-knockout mice displayed osteopenia. Because systemic Ggcx-knockout mice die shortly after birth from severe hemorrhage, the GGCX-mediated effect of vitamin K on bone metabolism has been difficult to evaluate. In this work, we utilized Ggcx-floxed mice to generate osteoblast-specific GGCX-deficient (Ggcx(Δobl/Δobl)) mice by crossing them with Col1-Cre mice. The bone mineral density (BMD) of Ggcx(Δobl/Δobl) mice was significantly higher than that of control Col1-Cre (Ggcx(+/+)) mice. Histomorphometrical analysis of trabecular bones in the proximal tibia showed increased osteoid volume and a higher rate of bone formation in Ggcx(Δobl/Δobl) mice. Histomorphometrical analysis of cortical bones revealed a thicker cortical width and a higher rate of bone formation in Ggcx(Δobl/Δobl) mice. Electron microscopic examination revealed disassembly of mineralized nodules and aberrant calcification of collagen fibers in Ggcx(Δobl/Δobl) mice. The mechanical properties of bones from Ggcx(Δobl/Δobl) mice tended to be stronger than those from control Ggcx(+/+) mice. These results suggest that GGCX in osteoblasts functions to prevent abnormal mineralization in bone formation, although this function may not be a prerequisite for the bone-protective effect of vitamin K. PMID:25600070

  9. Phylogenetic Analysis of Nucleus-Encoded Acetyl-CoA Carboxylases Targeted at the Cytosol and Plastid of Algae.

    KAUST Repository

    Huerlimann, Roger

    2015-07-01

    The understanding of algal phylogeny is being impeded by an unknown number of events of horizontal gene transfer (HGT), and primary and secondary/tertiary endosymbiosis. Through these events, previously heterotrophic eukaryotes developed photosynthesis and acquired new biochemical pathways. Acetyl-CoA carboxylase (ACCase) is a key enzyme in the fatty acid synthesis and elongation pathways in algae, where ACCase exists in two locations (cytosol and plastid) and in two forms (homomeric and heteromeric). All algae contain nucleus-encoded homomeric ACCase in the cytosol, independent of the origin of the plastid. Nucleus-encoded homomeric ACCase is also found in plastids of algae that arose from a secondary/tertiary endosymbiotic event. In contrast, plastids of algae that arose from a primary endosymbiotic event contain heteromeric ACCase, which consists of three nucleus-encoded and one plastid-encoded subunits. These properties of ACCase provide the potential to inform on the phylogenetic relationships of hosts and their plastids, allowing different hypothesis of endosymbiotic events to be tested. Alveolata (Dinoflagellata and Apicomplexa) and Chromista (Stramenopiles, Haptophyta and Cryptophyta) have traditionally been grouped together as Chromalveolata, forming the red lineage. However, recent genetic evidence groups the Stramenopiles, Alveolata and green plastid containing Rhizaria as SAR, excluding Haptophyta and Cryptophyta. Sequences coding for plastid and cytosol targeted homomeric ACCases were isolated from Isochrysis aff. galbana (TISO), Chromera velia and Nannochloropsis oculata, representing three taxonomic groups for which sequences were lacking. Phylogenetic analyses show that cytosolic ACCase strongly supports the SAR grouping. Conversely, plastidial ACCase groups the SAR with the Haptophyta, Cryptophyta and Prasinophyceae (Chlorophyta). These two ACCase based, phylogenetic relationships suggest that the plastidial homomeric ACCase was acquired by the

  10. Metabolic regulation of invadopodia and invasion by acetyl-CoA carboxylase 1 and de novo lipogenesis.

    Directory of Open Access Journals (Sweden)

    Kristen E N Scott

    Full Text Available Invadopodia are membrane protrusions that facilitate matrix degradation and cellular invasion. Although lipids have been implicated in several aspects of invadopodia formation, the contributions of de novo fatty acid synthesis and lipogenesis have not been defined. Inhibition of acetyl-CoA carboxylase 1 (ACC1, the committed step of fatty acid synthesis, reduced invadopodia formation in Src-transformed 3T3 (3T3-Src cells, and also decreased the ability to degrade gelatin. Inhibition of fatty acid synthesis through AMP-activated kinase (AMPK activation and ACC phosphorylation also decreased invadopodia incidence. The addition of exogenous 16∶0 and 18∶1 fatty acid, products of de novo fatty acid synthesis, restored invadopodia and gelatin degradation to cells with decreased ACC1 activity. Pharmacological inhibition of ACC also altered the phospholipid profile of 3T3-Src cells, with the majority of changes occurring in the phosphatidylcholine (PC species. Exogenous supplementation with the most abundant PC species, 34∶1 PC, restored invadopodia incidence, the ability to degrade gelatin and the ability to invade through matrigel to cells deficient in ACC1 activity. On the other hand, 30∶0 PC did not restore invadopodia and 36∶2 PC only restored invadopodia incidence and gelatin degradation, but not cellular invasion through matrigel. Pharmacological inhibition of ACC also reduced the ability of MDA-MB-231 breast, Snb19 glioblastoma, and PC-3 prostate cancer cells to invade through matrigel. Invasion of PC-3 cells through matrigel was also restored by 34∶1 PC supplementation. Collectively, the data elucidate the novel metabolic regulation of invadopodia and the invasive process by de novo fatty acid synthesis and lipogenesis.

  11. AMPK activation represses the human gene promoter of the cardiac isoform of acetyl-CoA carboxylase: Role of nuclear respiratory factor-1

    Energy Technology Data Exchange (ETDEWEB)

    Adam, Tasneem; Opie, Lionel H. [Hatter Cardiovascular Research Institute, Faculty of Health Sciences, University of Cape Town, Observatory 7925 (South Africa); Essop, M. Faadiel, E-mail: mfessop@sun.ac.za [Cardio-Metabolic Research Group (CMRG), Department of Physiological Sciences, Stellenbosch University, Stellenbosch 7600 (South Africa)

    2010-07-30

    Research highlights: {yields} AMPK inhibits acetyl-CoA carboxylase beta gene promoter activity. {yields} Nuclear respiratory factor-1 inhibits acetyl-CoA carboxylase beta promoter activity. {yields} AMPK regulates acetyl-CoA carboxylase beta at transcriptional level. -- Abstract: The cardiac-enriched isoform of acetyl-CoA carboxylase (ACC{beta}) produces malonyl-CoA, a potent inhibitor of carnitine palmitoyltransferase-1. AMPK inhibits ACC{beta} activity, lowering malonyl-CoA levels and promoting mitochondrial fatty acid {beta}-oxidation. Previously, AMPK increased promoter binding of nuclear respiratory factor-1 (NRF-1), a pivotal transcriptional modulator controlling gene expression of mitochondrial proteins. We therefore hypothesized that NRF-1 inhibits myocardial ACC{beta} promoter activity via AMPK activation. A human ACC{beta} promoter-luciferase construct was transiently transfected into neonatal cardiomyocytes {+-} a NRF-1 expression construct. NRF-1 overexpression decreased ACC{beta} gene promoter activity by 71 {+-} 4.6% (p < 0.001 vs. control). Transfections with 5'-end serial promoter deletions revealed that NRF-1-mediated repression of ACC{beta} was abolished with a pPII{beta}-18/+65-Luc deletion construct. AMPK activation dose-dependently reduced ACC{beta} promoter activity, while NRF-1 addition did not further decrease it. We also investigated NRF-1 inhibition in the presence of upstream stimulatory factor 1 (USF1), a known transactivator of the human ACC{beta} gene promoter. Here NRF-1 blunted USF1-dependent induction of ACC{beta} promoter activity by 58 {+-} 7.5% (p < 0.001 vs. control), reversed with a dominant negative NRF-1 construct. NRF-1 also suppressed endogenous USF1 transcriptional activity by 55 {+-} 6.2% (p < 0.001 vs. control). This study demonstrates that NRF-1 is a novel transcriptional inhibitor of the human ACC{beta} gene promoter in the mammalian heart. Our data extends AMPK regulation of ACC{beta} to the transcriptional level.

  12. Abundance and distribution of archaeal acetyl-CoA/propionyl-CoA carboxylase genes indicative for putatively chemoautotrophic Archaea in the tropical Atlantic's interior

    OpenAIRE

    Bergauer, Kristin; Sintes, Eva; van Bleijswijk, Judith; Witte, Harry; Herndl, Gerhard J.; Lueders, Tillmann

    2013-01-01

    Recently, evidence suggests that dark CO2 fixation in the pelagic realm of the ocean does not only occur in the suboxic and anoxic water bodies but also in the oxygenated meso- and bathypelagic waters of the North Atlantic. To elucidate the significance and phylogeny of the key organisms mediating dark CO2 fixation in the tropical Atlantic, we quantified functional genes indicative for CO2 fixation. We used a Q-PCR-based assay targeting the bifunctional acetyl-CoA/propionyl-CoA carboxylase (a...

  13. Protein phosphatases active on acetyl-CoA carboxylase phosphorylated by casein kinase I, casein kinase II and the cAMP-dependent protein kinase

    International Nuclear Information System (INIS)

    The protein phosphatases in rat liver cytosol, active on rat liver acetyl-CoA carboxylase (ACC) phosphorylated by casein kinase I, casein kinase II and the cAMP-dependent protein kinase, have been partially purified by anion-exchange and gel filtration chromatography. The major phosphatase activities against all three substrates copurify through fractionation and appear to be identical to protein phosphatases 2A1 and 2A2. No unique protein phosphatase active on 32P-ACC phosphorylated by the casein kinases was identified

  14. Influence of the nitrate concentration and source in the incorporation of 14CO2 by the RuBP-carboxylase from wheat (triticum aestivum) and maize (zea mays)

    International Nuclear Information System (INIS)

    The effect of the concentration and source of nitrogen in the culture media has been studied regarding its influence in the activity of the RuBP-carboxylase from wheat and maize during the first month of development. Wheat and maize has been chosen as plants representatives of two different types of CO2 assimilation: C3 and M- respectively. Plants have been grown in hydroponic media and under temperature, humidity and nutrient salts control. A negative effect of NH4 has been observed in the enzymatic activity of wheat seedlings, being this effect more remarkable as NH4 concentration increases and as long the time of treatment. In our experimental conditions the most favorable source of nitrogen has been N03NH4. The specific activity of the enzyme from wheat is about four times higher than in maize, even it decreases with time. This decreasing has not been observed in maize, with the exception of total absence of nitrogen in the media. We have not seen significant differences between the two photo periods which have been tested. Also, no differences have been found in the enzyme activities at the different NO3NH4 concentrations assayed, and it seems that RuBP-carboxylase metabolism is only affected in the case of absolute stress. (Author) 20 refs

  15. Antigen receptor-mediated regulation of sustained polyphosphoinositide turnover in a human T cell line. Evidence for a receptor-regulated pathway for production of phosphatidylinositol 4,5-bisphosphate

    International Nuclear Information System (INIS)

    Stimulation of the human T cell line, Jurkat, by the addition of monoclonal antibodies reactive with the T cell antigen receptor complex (CD3/Ti) leads to sustained increases in levels of inositol 1,4,5-trisphosphate. To investigate the possibility that the production of polyphosphoinositides is regulated during CD3/Ti stimulation, we studied Jurkat cells whose inositol phospholipids had been labeled to steady state with [3H]inositol, as well as Jurkat cells during nonequilibrium labeling with [32P]orthophosphate. The addition of CD3 monoclonal antibodies led to a 4-5-fold increase in [3H]inositol trisphosphate that was sustained for greater than 20 min. Within 60 s of CD3/Ti stimulation, [3H] phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) and [3H]phosphatidylinositol 4-phosphate (PtdIns4P) decreased by 65 and 35%, respectively. This change in [3H]PtdIns(4,5)P2 persisted for greater than 20 min. The decrease in [3H]PtdIns4P, however, was transient, and, after 5 min, the levels of [3H]PtdIns4P were comparable in stimulated and unstimulated cells. To examine the rate of flux through inositol phospholipids, we measured the CD3/Ti-stimulated changes in the ratio, 32P cpm/3H cpm, in each inositol phospholipid. CD3/Ti stimulation led to accelerated fluxes through PtdIns(4,5)P2 and phosphatidylinositol (PtdIns) that were maintained for greater than 20 min. After the initial 30 s, however, there was no detectable effect of anti-CD3 on flux through Ptsins4p. This observation suggested that, during CD3/Ti stimulation, production of PtdIns(4,5)P2 from PtdIns might occur via a small pool of PtdIns4P with a very high turnover. The existence of such a pool was established by determining that, in stimulated cells, the 32P-specific activity of the 1-position phosphate of PtdIns(4,5)P2 was 8-10-fold that of PtdIns4P

  16. Psychosine-triggered endomitosis is modulated by membrane sphingolipids through regulation of phosphoinositide 4,5-bisphosphate production at the cleavage furrow.

    Science.gov (United States)

    Watanabe, Hiroshi; Okahara, Kyohei; Naito-Matsui, Yuko; Abe, Mitsuhiro; Go, Shinji; Inokuchi, Jinichi; Okazaki, Toshiro; Kobayashi, Toshihide; Kozutsumi, Yasunori; Oka, Shogo; Takematsu, Hiromu

    2016-07-01

    Endomitosis is a special type of mitosis in which only cytokinesis-the final step of the cell division cycle-is defective, resulting in polyploid cells. Although endomitosis is biologically important, its regulatory aspects remain elusive. Psychosine, a lysogalactosylceramide, prevents proper cytokinesis when supplemented to proliferating cells. Cytokinetic inhibition by psychosine does not inhibit genome duplication. Consequently cells undergo multiple rounds of endomitotic cell cycles, resulting in the formation of giant multiploid cells. Here we successfully quantified psychosine-triggered multiploid cell formation, showing that membrane sphingolipids ratios modulate psychosine-triggered polyploidy in Namalwa cells. Among enzymes that experimentally remodel cellular sphingolipids, overexpression of glucosylceramide synthase to biosynthesize glycosylsphingolipids (GSLs) and neutral sphingomyelinase 2 to hydrolyze sphingomyelin (SM) additively enhanced psychosine-triggered multiploidy; almost all of the cells became polyploid. In the presence of psychosine, Namalwa cells showed attenuated cell surface SM clustering and suppression of phosphatidylinositol 4,5-bisphosphate production at the cleavage furrow, both important processes for cytokinesis. Depending on the sphingolipid balance between GSLs and SM, Namalwa cells could be effectively converted to viable multiploid cells with psychosine. PMID:27170180

  17. The interaction between the pleckstrin homology domain of ceramide kinase and phosphatidylinositol 4,5-bisphosphate regulates the plasma membrane targeting and ceramide 1-phosphate levels

    International Nuclear Information System (INIS)

    Ceramide kinase (CERK) converts ceramide to ceramide-1-phosphate (C1P), which has recently emerged as a new bioactive molecule capable of regulating diverse cellular functions. The N-terminus of the CERK protein encompasses a sequence motif known as a pleckstrin homology (PH) domain. Although the PH domain was previously demonstrated to be an important domain for the subcellular localization of CERK, the precise properties of this domain remained unclear. In this study, we reveal that the PH domain of CERK exhibits high affinity for phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), among other lipids. Furthermore, in COS7 cells, GFP-fused CERK translocated rapidly from the cytoplasm to the plasma membrane in response to hyper-osmotic stress, which is known to increase the intracellular PI(4,5)P2 levels, whereas a PH domain deletion mutant did not. Additionally, in [32P]orthophosphate-labeled COS7 cells, the translocation of CERK to the plasma membrane induced a 2.8-fold increase in C1P levels. The study presented here provides insight into the crucial role of the CERK-PH domain in plasma membrane targeting, through its binding to PI(4,5)P2, and subsequent induction of C1P production in the vicinity of the membrane

  18. Photoperiodism and crassulacean acid metabolism : I. Immunological and kinetic evidences for different patterns of phosphoenolpyruvate carboxylase isoforms in photoperiodically inducible and non-inducible Crassulacean acid metabolism plants.

    Science.gov (United States)

    Brulfert, J; Müller, D; Kluge, M; Queiroz, O

    1982-05-01

    Plants of Kalanchoe blossfeldiana v. Poelln. Tom Thumb and Sedum morganianum E. Walth. were grown under controlled photoperiodic conditions under either short or long days. Gaz exchange measurements confirmed that in K. blossfeldiana Crassulacean acid metabolism (CAM) was photoperiodically inducible and that S. morganianum performed CAM independently of photoperiod. With K. blossfeldiana, a comparison of catalytic and regulatory properties of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) from short-day and long-day grown plants showed differences, but not with S. morganianum. Ouchterlony double diffusion tests and immunotitration experiments (using a S. morganianum PEPC antibody) established that CAM is induced in K. blossfeldiana-but not in S. morganianum-through the synthesis of a new PEPC isoform; this form shows an immunological behavior different from that prevailing under non-inductive conditions and can be considered as specific for CAM performance. PMID:24276159

  19. Accumulation fatty acids of in Chlorella vulgaris under heterotrophic conditions in relation to activity of acetyl-CoA carboxylase, temperature, and co-immobilization with Azospirillum brasilense

    Science.gov (United States)

    Leyva, Luis A.; Bashan, Yoav; Mendoza, Alberto; de-Bashan, Luz E.

    2014-10-01

    The relation between fatty acid accumulation, activity of acetyl-CoA carboxylase (ACC), and consequently lipid accumulation was studied in the microalgae Chlorella vulgaris co-immobilized with the plant growth-promoting bacterium Azospirillum brasilense under dark heterotrophic conditions with Na acetate as a carbon source. In C. vulgaris immobilized alone, cultivation experiments for 6 days showed that ACC activity is directly related to fatty acid accumulation, especially in the last 3 days. In co-immobilization experiments, A. brasilense exerted a significant positive effect over ACC activity, increased the quantity in all nine main fatty acids, increased total lipid accumulation in C. vulgaris, and mitigated negative effects of nonoptimal temperature for growth. No correlation between ACC activity and lipid accumulation in the cells was established for three different temperatures. This study demonstrated that the interaction between A. brasilense and C. vulgaris has a significant effect on fatty acid and lipid accumulation in the microalgae.

  20. Genes encoding the alpha-carboxyltransferase subunit of acetyl-CoA carboxylase from Brassica napus and parental species: cloning, expression patterns, and evolution.

    Science.gov (United States)

    Li, Zhi-Guo; Yin, Wei-Bo; Guo, Huan; Song, Li-Ying; Chen, Yu-Hong; Guan, Rong-Zhan; Wang, Jing-Qiao; Wang, Richard R-C; Hu, Zan-Min

    2010-05-01

    Heteromeric acetyl coenzyme A carboxylase (ACCase), a rate-limiting enzyme in fatty acid biosynthesis in dicots, is a multi-enzyme complex consisting of biotin carboxylase, biotin carboxyl carrier protein, and carboxyltransferase (alpha-CT and beta-CT). In the present study, four genes encoding alpha-CT were cloned from Brassica napus, and two were cloned from each of the two parental species, B. rapa and B. oleracea. Comparative and cluster analyses indicated that these genes were divided into two major groups. The major divergence between group-1 and group-2 occurred in the second intron. Group-2 alpha-CT genes represented the ancestral form in the genus Brassica. The divergence of group-1 and group-2 genes occurred in their common ancestor 12.96-17.78 million years ago (MYA), soon after the divergence of Arabidopsis thaliana and Brassica (15-20 MYA). This time of divergence is identical to that reported for the paralogous subgenomes of diploid Brassica species (13-17 MYA). Real-time reverse transcription PCR revealed that the expression patterns of the two groups of genes were similar in different organs, except in leaves. To better understand the regulation and evolution of alpha-CT genes, promoter regions from two sets of orthologous gene copies from B. napus, B. rapa, and B. oleracea were cloned and compared. The function of the promoter of gene Bnalpha-CT-1-1 in group-1 and gene Bnalpha-CT-2-1 in group-2 was examined by assaying beta-glucuronidase activity in transgenic A. thaliana. Our results will be helpful in elucidating the evolution and regulation of ACCase in oilseed rape. PMID:20616867

  1. Cloning and Characterization of a Pyruvate Carboxylase Gene from Penicillium rubens and Overexpression of the Genein the Yeast Yarrowia lipolytica for Enhanced Citric Acid Production.

    Science.gov (United States)

    Fu, Ge-Yi; Lu, Yi; Chi, Zhe; Liu, Guang-Lei; Zhao, Shou-Feng; Jiang, Hong; Chi, Zhen-Ming

    2016-02-01

    In this study, a pyruvate carboxylase gene (PYC1) from a marine fungus Penicillium rubens I607 was cloned and characterized. ORF of the gene (accession number: KM397349.1) had 3534 bp encoding 1177 amino acids with a molecular weight of 127.531 kDa and a PI of 6.20. The promoter of the gene was located at -1200 bp and contained a TATAA box, several CAAT boxes and a sequence 5'-SYGGRG-3'. The PYC1 deduced from the gene had no signal peptide, was a homotetramer (α4), and had the four functional domains. After expression of the PYC1 gene from the marine fungus in the marine-derived yeast Yarrowia lipolytica SWJ-1b, the transformant PR32 obtained had much higher specific pyruvate carboxylase activity (0.53 U/mg) than Y. lipolytica SWJ-1b (0.07 U/mg), and the PYC1 gene expression (133.8%) and citric acid production (70.2 g/l) by the transformant PR32 were also greatly enhanced compared to those (100 % and 27.3 g/l) by Y. lipolytica SWJ-1b. When glucose concentration in the medium was 60.0 g/l, citric acid (CA) concentration formed by the transformant PR32 was 36.1 g/l, leading to conversion of 62.1% of glucose into CA. During a 10-l fed-batch fermentation, the final concentration of CA was 111.1 ± 1.3 g/l, the yield was 0.93 g/g, the productivity was 0.46 g/l/h, and only 1.72 g/l reducing sugar was left in the fermented medium within 240 h. HPLC analysis showed that most of the fermentation products were CA. However, minor malic acid and other unknown products also existed in the culture. PMID:26470708

  2. New insights into the post-translational modification of multiple phosphoenolpyruvate carboxylase isoenzymes by phosphorylation and monoubiquitination during sorghum seed development and germination.

    Science.gov (United States)

    Ruiz-Ballesta, Isabel; Baena, Guillermo; Gandullo, Jacinto; Wang, Liqun; She, Yi-Min; Plaxton, William Charles; Echevarría, Cristina

    2016-05-01

    Phosphoenolpyruvate carboxylase (PEPC; E.C. 4.1.1.31) was characterized in developing and germinating sorghum seeds, focusing on the transcript and polypeptide abundance of multiple plant-type phosphoenolpyruvate carboxylase (PTPC) genes, and the post-translational modification of each isoenzyme by phosphorylation versus monoubiquitination during germination. We observed high levels of SbPPC4 (Sb07g014960) transcripts during early development (stage I), and extensive transcript abundance of SbPPC2 (Sb02g021090) and SbPPC3 (Sb04g008720) throughout the entire life cycle of the seed. Although tandem mass spectrometry (MS) analysis of immunopurified PTPC indicated that four different PTPC isoenzymes were expressed in the developing and germinating seeds, SbPPC3 was the most abundant isozyme of the developing seed, and of the embryo and the aleurone layer of germinating seeds. In vivo phosphorylation of the different PTPC isoenzymes at their conserved N-terminal seryl phosphorylation site during germination was also established by MS/MS analysis. Furthermore, three of the four isoenzymes were partially monoubiquitinated, with MS/MS pinpointing SbPPC2 and SbPPC3 monoubiquitination at the conserved Lys-630 and Lys-624 residues, respectively. Our results demonstrate that monoubiquitination and phosphorylation simultaneously occur in vivo with different PTPC isozymes during seed germination. In addition, we show that PTPC monoubiquitination in germinating sorghum seeds always increases at stage II (emergence of the radicle), is maintained during the aerobic period of rapid cell division and reserve mobilization, and remains relatively constant until stage IV-V when coleoptiles initiate the formation of the photosynthetic tissues. PMID:27194739

  3. Mutations in Nature Conferred a High Affinity Phosphatidylinositol 4,5-Bisphosphate-binding Site in Vertebrate Inwardly Rectifying Potassium Channels.

    Science.gov (United States)

    Tang, Qiong-Yao; Larry, Trevor; Hendra, Kalen; Yamamoto, Erica; Bell, Jessica; Cui, Meng; Logothetis, Diomedes E; Boland, Linda M

    2015-07-01

    All vertebrate inwardly rectifying potassium (Kir) channels are activated by phosphatidylinositol 4,5-bisphosphate (PIP2) (Logothetis, D. E., Petrou, V. I., Zhang, M., Mahajan, R., Meng, X. Y., Adney, S. K., Cui, M., and Baki, L. (2015) Annu. Rev. Physiol. 77, 81-104; Fürst, O., Mondou, B., and D'Avanzo, N. (2014) Front. Physiol. 4, 404-404). Structural components of a PIP2-binding site are conserved in vertebrate Kir channels but not in distantly related animals such as sponges and sea anemones. To expand our understanding of the structure-function relationships of PIP2 regulation of Kir channels, we studied AqKir, which was cloned from the marine sponge Amphimedon queenslandica, an animal that represents the phylogenetically oldest metazoans. A requirement for PIP2 in the maintenance of AqKir activity was examined in intact oocytes by activation of a co-expressed voltage-sensing phosphatase, application of wortmannin (at micromolar concentrations), and activation of a co-expressed muscarinic acetylcholine receptor. All three mechanisms to reduce the availability of PIP2 resulted in inhibition of AqKir current. However, time-dependent rundown of AqKir currents in inside-out patches could not be re-activated by direct application to the inside membrane surface of water-soluble dioctanoyl PIP2, and the current was incompletely re-activated by the more hydrophobic arachidonyl stearyl PIP2. When we introduced mutations to AqKir to restore two positive charges within the vertebrate PIP2-binding site, both forms of PIP2 strongly re-activated the mutant sponge channels in inside-out patches. Molecular dynamics simulations validate the additional hydrogen bonding potential of the sponge channel mutants. Thus, nature's mutations conferred a high affinity activation of vertebrate Kir channels by PIP2, and this is a more recent evolutionary development than the structures that explain ion channel selectivity and inward rectification. PMID:25957411

  4. Plasma membrane phosphatidylinositol 4,5 bisphosphate is required for internalization of foot-and-mouth disease virus and vesicular stomatitis virus.

    Directory of Open Access Journals (Sweden)

    Angela Vázquez-Calvo

    Full Text Available Phosphatidylinositol-4,5-bisphosphate, PI(4,5P(2, is a phospholipid which plays important roles in clathrin-mediated endocytosis. To investigate the possible role of this lipid on viral entry, two viruses important for animal health were selected: the enveloped vesicular stomatitis virus (VSV - which uses a well characterized clathrin mediated endocytic route - and two different variants of the non-enveloped foot-and-mouth disease virus (FMDV with distinct receptor specificities. The expression of a dominant negative dynamin, a PI(4,5P(2 effector protein, inhibited the internalization and infection of VSV and both FMDV isolates. Depletion of PI(4,5P(2 from plasma membrane using ionomycin or an inducible system, and inhibition of its de novo synthesis with 1-butanol revealed that VSV as well as FMDV C-S8c1, which uses integrins as receptor, displayed a high dependence on PI(4,5P(2 for internalization. Expression of a kinase dead mutant (KD of phosphatidylinositol-4-phosphate-5-kinase Iα (PIP5K-Iα, an enzyme responsible for PI(4,5P(2 synthesis that regulates clathrin-dependent endocytosis, also impaired entry and infection of VSV and FMDV C-S8c1. Interestingly FMDV MARLS variant that uses receptors other than integrins for cell entry was less sensitive to PI(4,5P(2 depletion, and was not inhibited by the expression of the KD PIP5K-Iα mutant suggesting the involvement of endocytic routes other than the clathrin-mediated on its entry. These results highlight the role of PI(4,5P(2 and PIP5K-Iα on clathrin-mediated viral entry.

  5. Ca2+ Influx through Store-operated Calcium Channels Replenishes the Functional Phosphatidylinositol 4,5-Bisphosphate Pool Used by Cysteinyl Leukotriene Type I Receptors.

    Science.gov (United States)

    Alswied, Abdullah; Parekh, Anant B

    2015-12-01

    Oscillations in cytoplasmic Ca(2+) concentration are a universal mode of signaling following physiological levels of stimulation with agonists that engage the phospholipase C pathway. Sustained cytoplasmic Ca(2+) oscillations require replenishment of the membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PIP2), the source of the Ca(2+)-releasing second messenger inositol trisphosphate. Here we show that cytoplasmic Ca(2+) oscillations induced by cysteinyl leukotriene type I receptor activation run down when cells are pretreated with Li(+), an inhibitor of inositol monophosphatases that prevents PIP2 resynthesis. In Li(+)-treated cells, cytoplasmic Ca(2+) signals evoked by an agonist were rescued by addition of exogenous inositol or phosphatidylinositol 4-phosphate (PI4P). Knockdown of the phosphatidylinositol 4-phosphate 5 (PIP5) kinases α and γ resulted in rapid loss of the intracellular Ca(2+) oscillations and also prevented rescue by PI4P. Knockdown of talin1, a protein that helps regulate PIP5 kinases, accelerated rundown of cytoplasmic Ca(2+) oscillations, and these could not be rescued by inositol or PI4P. In Li(+)-treated cells, recovery of the cytoplasmic Ca(2+) oscillations in the presence of inositol or PI4P was suppressed when Ca(2+) influx through store-operated Ca(2+) channels was inhibited. After rundown of the Ca(2+) signals following leukotriene receptor activation, stimulation of P2Y receptors evoked prominent inositol trisphosphate-dependent Ca(2+) release. Therefore, leukotriene and P2Y receptors utilize distinct membrane PIP2 pools. Our findings show that store-operated Ca(2+) entry is needed to sustain cytoplasmic Ca(2+) signaling following leukotriene receptor activation both by refilling the Ca(2+) stores and by helping to replenish the PIP2 pool accessible to leukotriene receptors, ostensibly through control of PIP5 kinase activity. PMID:26468289

  6. Characterization of the S. cerevisiae inp51 mutant links phosphatidylinositol 4,5-bisphosphate levels with lipid content, membrane fluidity and cold growth.

    Science.gov (United States)

    Córcoles-Sáez, Isaac; Hernández, Maria Luisa; Martínez-Rivas, Jose Manuel; Prieto, Jose A; Randez-Gil, Francisca

    2016-03-01

    Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] and its derivatives diphosphoinositol phosphates (DPIPs) play key signaling and regulatory roles. However, a direct function of these molecules in lipid and membrane homeostasis remains obscure. Here, we have studied the cold tolerance phenotype of yeast cells lacking the Inp51-mediated phosphoinositide-5-phosphatase. Genetic and biochemical approaches showed that increased metabolism of PI(4,5)P2 reduces the activity of the Pho85 kinase by increasing the levels of the DPIP isomer 1-IP7. This effect was key in the cold tolerance phenotype. Indeed, pho85 mutant cells grew better than the wild-type at 15 °C, and lack of this kinase abolished the inp51-mediated cold phenotype. Remarkably, reduced Pho85 function by loss of Inp51 affected the activity of the Pho85-regulated target Pah1, the yeast phosphatidate phosphatase. Cells lacking Inp51 showed reduced Pah1 abundance, derepression of an INO1-lacZ reporter, decreased content of triacylglycerides and elevated levels of phosphatidate, hallmarks of the pah1 mutant. However, the inp51 phenotype was not associated to low Pah1 activity since deletion of PAH1 caused cold sensitivity. In addition, the inp51 mutant exhibited features not shared by pah1, including a 40%-reduction in total lipid content and decreased membrane fluidity. These changes may influence the activity of membrane-anchored and/or associated proteins since deletion of INP51 slows down the transit to the vacuole of the fluorescent dye FM4-64. In conclusion, our work supports a model in which changes in the PI(4,5)P2 pool affect the 1-IP7 levels modulating the activity of Pho85, Pah1 and likely additional Pho85-controlled targets, and regulate lipid composition and membrane properties. PMID:26724696

  7. Multiple sources of 1,2-diacylglycerol in isolated rat pancreatic acini stimulated by cholecystokinin. Involvement of phosphatidylinositol bisphosphate and phosphatidylcholine hydrolysis

    International Nuclear Information System (INIS)

    Changes in the cellular content of 1,2-diacylglycerol (DAG) in isolated rat pancreatic acini in response to agonist stimulation were studied using a sensitive mass assay. When acini were stimulated by 10 nM COOH-terminal cholecystokinin-octapeptide (CCK8), the increase in DAG was biphasic, consisting of an early peak at 5 s and a second, larger, gradual increase that was maximal by 15 min. The basal level of DAG in acini was 1.04 nmol/mg of protein, which was increased to 1.24 nmol/mg of protein at 5 s and 2.76 nmol/mg of protein at 30 min. In comparison, the increase in DAG stimulated by 30 pM CCK8, a submaximal concentration for amylase release, was monophasic, increasing without an early peak but sustained to 60 min. Other Ca2+-mobilizing secretagogues such as carbamylcholine and bombesin increased DAG in acini, whereas vasoactive intestinal peptide, which acts to increase cAMP, had no effect. Phorbol ester and Ca2+ ionophore also stimulated DAG production. Analysis of the mass level of inositol 1,4,5-trisphosphate (1,4,5-IP3) showed that the generation of 1,4,5-IP3 stimulated by 10 nM CCK8 peaked at 5 s, a finding consistent with the early peak of DAG. The basal level was 4.7 pmol/mg of protein, which was increased to 144.6 pmol/mg of protein at 5 s by 10 nM CCK8. The levels of 1,4,5-IP3 then returned toward basal in contrast to the gradual and sustained increase of DAG. The dose dependencies of 1,4,5-IP3 and DAG formation at 5 s with respect to CCK8 were almost identical. This suggests that phosphatidylinositol 4,5-bisphosphate hydrolysis is a major source of the early increase in DAG but not of the sustained increase in DAG. Therefore, a possible contribution of phosphatidylcholine hydrolysis to DAG formation was examined utilizing acini prelabeled with [3H]choline. CCK8 (1 nM) maximally increased [3H]choline metabolite release by 133% of control at 30 min

  8. The glossyhead1 Allele of ACC1 Reveals a Principal Role for Multidomain Acetyl-Coenzyme A Carboxylase in the Biosynthesis of Cuticular Waxes by Arabidopsis

    Energy Technology Data Exchange (ETDEWEB)

    Lu, S.; Xu, C.; Zhao, H.; Parsons, E. P.; Kosma, D. K.; Xu, X.; Chao, D.; Lohrey, G.; Bangarusamy, D. K.; Wang, G.; Bressan, R. A.; Jenks, M. A.

    2011-11-01

    A novel mutant of Arabidopsis (Arabidopsis thaliana), having highly glossy inflorescence stems, postgenital fusion in floral organs, and reduced fertility, was isolated from an ethyl methanesulfonate-mutagenized population and designated glossyhead1 (gsd1). The gsd1 locus was mapped to chromosome 1, and the causal gene was identified as a new allele of Acetyl-Coenzyme A Carboxylase1 (ACC1), a gene encoding the main enzyme in cytosolic malonyl-coenzyme A synthesis. This, to our knowledge, is the first mutant allele of ACC1 that does not cause lethality at the seed or early germination stage, allowing for the first time a detailed analysis of ACC1 function in mature tissues. Broad lipid profiling of mature gsd1 organs revealed a primary role for ACC1 in the biosynthesis of the very-long-chain fatty acids (C{sub 20:0} or longer) associated with cuticular waxes and triacylglycerols. Unexpectedly, transcriptome analysis revealed that gsd1 has limited impact on any lipid metabolic networks but instead has a large effect on environmental stress-responsive pathways, especially senescence and ethylene synthesis determinants, indicating a possible role for the cytosolic malonyl-coenzyme A-derived lipids in stress response signaling.

  9. Decreasing the Rate of Metabolic Ketone Reduction in the Discovery of a Clinical Acetyl-CoA Carboxylase Inhibitor for the Treatment of Diabetes

    Energy Technology Data Exchange (ETDEWEB)

    Griffith, David A. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Kung, Daniel W. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Esler, William P. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Amor, Paul A. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Bagley, Scott W. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Beysen, Carine [KineMed Inc., Emeryville, CA (United States); Carvajal-Gonzalez, Santos [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Doran, Shawn D. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Limberakis, Chris [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Mathiowetz, Alan M. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); McPherson, Kirk [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Price, David A. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Ravussin, Eric [Louisiana State Univ., Baton Rouge, LA (United States); Sonnenberg, Gabriele E. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Southers, James A. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Sweet, Laurel J. [Pfizer Worldwide Research and Development, Cambridge, MA (United States); Turner, Scott M. [KineMed Inc., Emeryville, CA (United States); Vajdos, Felix F. [Pfizer Worldwide Research and Development, Cambridge, MA (United States)

    2014-12-26

    We found that Acetyl-CoA carboxylase (ACC) inhibitors offer significant potential for the treatment of type 2 diabetes mellitus (T2DM), hepatic steatosis, and cancer. However, the identification of tool compounds suitable to test the hypothesis in human trials has been challenging. An advanced series of spirocyclic ketone-containing ACC inhibitors recently reported by Pfizer were metabolized in vivo by ketone reduction, which complicated human pharmacology projections. Here, we disclose that this metabolic reduction can be greatly attenuated through introduction of steric hindrance adjacent to the ketone carbonyl. Incorporation of weakly basic functionality improved solubility and led to the identification of 9 as a clinical candidate for the treatment of T2DM. Phase I clinical studies demonstrated dose-proportional increases in exposure, single-dose inhibition of de novo lipogenesis (DNL), and changes in indirect calorimetry consistent with increased whole-body fatty acid oxidation. This demonstration of target engagement validates the use of compound 9 to evaluate the role of DNL in human disease.

  10. The glossyhead1 allele of acc1 reveals a principal role for multidomain acetyl-coenzyme a carboxylase in the biosynthesis of cuticular waxes by Arabidopsis

    KAUST Repository

    Lu, Shiyou

    2011-09-23

    A novel mutant of Arabidopsis (Arabidopsis thaliana), having highly glossy inflorescence stems, postgenital fusion in floral organs, and reduced fertility, was isolated from an ethyl methanesulfonate-mutagenized population and designated glossyhead1 (gsd1). The gsd1 locus was mapped to chromosome 1, and the causal gene was identified as a new allele of Acetyl-Coenzyme A Carboxylase1 (ACC1), a gene encoding the main enzyme in cytosolic malonyl-coenzyme A synthesis. This, to our knowledge, is the first mutant allele of ACC1 that does not cause lethality at the seed or early germination stage, allowing for the first time a detailed analysis of ACC1 function in mature tissues. Broad lipid profiling of mature gsd1 organs revealed a primary role for ACC1 in the biosynthesis of the very-long-chain fatty acids (C 20:0 or longer) associated with cuticular waxes and triacylglycerols. Unexpectedly, transcriptome analysis revealed that gsd1 has limited impact on any lipid metabolic networks but instead has a large effect on environmental stress-responsive pathways, especially senescence and ethylene synthesis determinants, indicating a possible role for the cytosolic malonyl-coenzyme A-derived lipids in stress response signaling. © 2011 American Society of Plant Biologists. All Rights Reserved.

  11. Biotin augments acetyl CoA carboxylase 2 gene expression in the hypothalamus, leading to the suppression of food intake in mice.

    Science.gov (United States)

    Sone, Hideyuki; Kamiyama, Shin; Higuchi, Mutsumi; Fujino, Kaho; Kubo, Shizuka; Miyazawa, Masami; Shirato, Saya; Hiroi, Yuka; Shiozawa, Kota

    2016-07-29

    It is known that biotin prevents the development of diabetes by increasing the functions of pancreatic beta-cells and improving insulin sensitivity in the periphery. However, its anti-obesity effects such as anorectic effects remain to be clarified. Acetyl CoA carboxylase (ACC), a biotin-dependent enzyme, has two isoforms (ACC1 and ACC2) and serves to catalyze the reaction of acetyl CoA to malonyl CoA. In the hypothalamus, ACC2 increases the production of malonyl CoA, which acts as a satiety signal. In this study, we investigated whether biotin increases the gene expression of ACC2 in the hypothalamus and suppresses food intake in mice administered excessive biotin. Food intake was significantly decreased by biotin, but plasma regulators of appetite, including glucose, ghrelin, and leptin, were not affected. On the other hand, biotin notably accumulated in the hypothalamus and enhanced ACC2 gene expression there, but it did not change the gene expression of ACC1, malonyl CoA decarboxylase (a malonyl CoA-degrading enzyme), and AMP-activated protein kinase α-2 (an ACC-inhibitory enzyme). These findings strongly suggest that biotin potentiates the suppression of appetite by upregulating ACC2 gene expression in the hypothalamus. This effect of biotin may contribute to the prevention of diabetes by biotin treatment. PMID:27181349

  12. Search for novel targets of the PII signal transduction protein in Bacteria identifies the BCCP component of acetyl-CoA carboxylase as a PII binding partner.

    Science.gov (United States)

    Rodrigues, Thiago E; Gerhardt, Edileusa C M; Oliveira, Marco A; Chubatsu, Leda S; Pedrosa, Fabio O; Souza, Emanuel M; Souza, Gustavo A; Müller-Santos, Marcelo; Huergo, Luciano F

    2014-02-01

    The PII family comprises a group of widely distributed signal transduction proteins. The archetypal function of PII is to regulate nitrogen metabolism in bacteria. As PII can sense a range of metabolic signals, it has been suggested that the number of metabolic pathways regulated by PII may be much greater than described in the literature. In order to provide experimental evidence for this hypothesis a PII protein affinity column was used to identify PII targets in Azospirillum brasilense. One of the PII partners identified was the biotin carboxyl carrier protein (BCCP), a component of the acetyl-CoA carboxylase which catalyses the committed step in fatty acid biosynthesis. As BCCP had been previously identified as a PII target in Arabidopsis thaliana we hypothesized that the PII -BCCP interaction would be conserved throughout Bacteria. In vitro experiments using purified proteins confirmed that the PII -BCCP interaction is conserved in Escherichia coli. The BCCP-PII interaction required MgATP and was dissociated by increasing 2-oxoglutarate. The interaction was modestly affected by the post-translational uridylylation status of PII ; however, it was completely dependent on the post-translational biotinylation of BCCP. PMID:24329683

  13. Bacterial Hydrolysis of Protein and Methylated Protein and Its Implications for Studies of Protein Degradation in Aquatic Systems

    OpenAIRE

    Keil, Richard G.; Kirchman, David L.

    1992-01-01

    Ribulose 1,5-bisphosphate carboxylase was radiolabelled by in vitro translation, resulting in uniformly labelled ribulose 1,5-bisphosphate carboxylase, and also by reductive methylation. We investigated the degradation of the two forms of radiolabelled protein by natural bacterial populations. Although total hydrolysis of uniformly labelled protein and methylated protein was nearly equal, percent assimilation, respiration, and release as low-molecular-weight material were different. Radioacti...

  14. Inhibition of acetyl-CoA carboxylases by soraphen A prevents lipid accumulation and adipocyte differentiation in 3T3-L1 cells.

    Science.gov (United States)

    Cordonier, Elizabeth L; Jarecke, Sarah K; Hollinger, Frances E; Zempleni, Janos

    2016-06-01

    Acetyl-CoA carboxylases (ACC) 1 and 2 catalyze the carboxylation of acetyl-CoA to malonyl-CoA and depend on biotin as a coenzyme. ACC1 localizes in the cytoplasm and produces malonyl-CoA for fatty acid (FA) synthesis. ACC2 localizes in the outer mitochondrial membrane and produces malonyl-CoA that inhibits FA import into mitochondria for subsequent oxidation. We hypothesized that ACCs are checkpoints in adipocyte differentiation and tested this hypothesis using the ACC1 and ACC2 inhibitor soraphen A (SA) in murine 3T3-L1 preadipocytes. When 3T3-L1 cells were treated with 100nM SA for 8 days after induction of differentiation, the expression of PPARγ mRNA and FABP4 mRNA decreased by 40% and 50%, respectively, compared with solvent controls; the decrease in gene expression was accompanied by a decrease in FABP4 protein expression and associated with a decrease in lipid droplet accumulation. The rate of FA oxidation was 300% greater in SA-treated cells compared with vehicle controls. Treatment with exogenous palmitate restored PPARγ and FABP4 mRNA expression and FABP4 protein expression in SA-treated cells. In contrast, SA did not alter lipid accumulation if treatment was initiated on day eight after induction of differentiation. We conclude that loss of ACC1-dependent FA synthesis and loss of ACC2-dependent inhibition of FA oxidation prevent lipid accumulation in adipocytes and inhibit early stages of adipocyte differentiation. PMID:27041646

  15. Discrimination in the dark. Resolving the interplay between metabolic and physical constraints to phosphoenolpyruvate carboxylase activity during the crassulacean acid metabolism cycle.

    Science.gov (United States)

    Griffiths, Howard; Cousins, Asaph B; Badger, Murray R; von Caemmerer, Susanne

    2007-02-01

    A model defining carbon isotope discrimination (delta13C) for crassulacean acid metabolism (CAM) plants was experimentally validated using Kalanchoe daigremontiana. Simultaneous measurements of gas exchange and instantaneous CO2 discrimination (for 13C and 18O) were made from late photoperiod (phase IV of CAM), throughout the dark period (phase I), and into the light (phase II). Measurements of CO2 response curves throughout the dark period revealed changing phosphoenolpyruvate carboxylase (PEPC) capacity. These systematic changes in PEPC capacity were tracked by net CO2 uptake, stomatal conductance, and online delta13C signal; all declined at the start of the dark period, then increased to a maximum 2 h before dawn. Measurements of delta13C were higher than predicted from the ratio of intercellular to external CO2 (p(i)/p(a)) and fractionation associated with CO2 hydration and PEPC carboxylations alone, such that the dark period mesophyll conductance, g(i), was 0.044 mol m(-2) s(-1) bar(-1). A higher estimate of g(i) (0.085 mol m(-2) s(-1) bar(-1)) was needed to account for the modeled and measured delta18O discrimination throughout the dark period. The differences in estimates of g(i) from the two isotope measurements, and an offset of -5.5 per thousand between the 18O content of source and transpired water, suggest spatial variations in either CO2 diffusion path length and/or carbonic anhydrase activity, either within individual cells or across a succulent leaf. Our measurements support the model predictions to show that internal CO2 diffusion limitations within CAM leaves increase delta13C discrimination during nighttime CO2 fixation while reducing delta13C during phase IV. When evaluating the phylogenetic distribution of CAM, carbon isotope composition will reflect these diffusive limitations as well as relative contributions from C3 and C4 biochemistry. PMID:17142488

  16. Computational simulations of structural role of the active-site W374C mutation of acetyl-coenzyme-A carboxylase: multi-drug resistance mechanism.

    Science.gov (United States)

    Zhu, Xiao-Lei; Yang, Wen-Chao; Yu, Ning-Xi; Yang, Sheng-Gang; Yang, Guang-Fu

    2011-03-01

    Herbicides targeting grass plastidic acetyl-CoA carboxylase (ACCase, EC 6.4.1.2) are selectively effective against graminicides. The intensive worldwide use of this herbicide family has selected for resistance genes in a number of grass weed species. Recently, the active-site W374C mutation was found to confer multi-drug resistance toward haloxyfop (HF), fenoxaprop (FR), Diclofop (DF), and clodinafop (CF) in A. myosuroides. In order to uncover the resistance mechanism due to W374C mutation, the binding of above-mentioned four herbicides to both wild-type and the mutant-type ACCase was investigated in the current work by molecular docking and molecular dynamics (MD) simulations. The binding free energies were calculated by molecular mechanics-Poisson-Boltzmann surface area (MM/PBSA) method. The calculated binding free energy values for four herbicides were qualitatively consistent with the experimental order of IC(50) values. All the computational model and energetic results indicated that the W374C mutation has great effects on the conformational change of the binding pocket and the ligand-protein interactions. The most significant conformational change was found to be associated with the aromatic amino acid residues, such as Phe377, Tyr161' and Trp346. As a result, the π-π interaction between the ligand and the residue of Phe377 and Tyr161', which make important contributions to the binding affinity, was decreased after mutation and the binding affinity for the inhibitors to the mutant-type ACCase was less than that to the wild-type enzyme, which accounts for the molecular basis of herbicidal resistance. The structural role and mechanistic insights obtained from computational simulations will provide a new starting point for the rational design of novel inhibitors to overcome drug resistance associated with W374C mutation. PMID:20499260

  17. Expression and methylation of microsomal triglyceride transfer protein and acetyl-CoA carboxylase are associated with fatty liver syndrome in chicken.

    Science.gov (United States)

    Liu, Zhen; Li, Qinghe; Liu, Ranran; Zhao, Guiping; Zhang, Yonghong; Zheng, Maiqing; Cui, Huanxian; Li, Peng; Cui, Xiaoyan; Liu, Jie; Wen, Jie

    2016-06-01

    The typical characteristic of fatty liver syndrome (FLS) is an increased hepatic triacylglycerol content, and a sudden decline in egg production often occurs. FLS may develop into fatty liver hemorrhagic syndrome (FLHS), characterized by sudden death from hepatic rupture and hemorrhage. DNA methylation is associated with transcriptional silencing, leading to the etiology and pathogenesis of some animal diseases. The roles of DNA methylation in the genesis of FLS, however, are largely unknown. The lipogenic methyl-deficient diet (MDD) caused FLS similar to human nonalcoholic steatohepatitis (NASH). After 16 Jingxing-Huang (JXH) hens were fed MDD for 10 wk, eight exhibited FLS (designated as FLS-susceptible birds); the remainder, without FLS, served as controls (NFLS). Physiological and biochemical variables, gene expression levels, and DNA methylation were determined in the liver. The development of FLS in JXH hens was accompanied by abnormal lipid accumulation. Relative expression of acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and microsomal triglyceride transfer protein (MTTP) were significantly up-regulated in the FLS group in comparison with the NFLS group. The transcript abundance of sterol regulatory element binding protein 1 (SREBP-1c), stearoyl-CoA desaturase (SCD), liver X receptor alpha (LXRα), peroxisome proliferator-activated receptor alpha (PPARα), and peroxisome proliferator-activated receptor gamma (PPARγ) did not differ between the two groups. Interestingly, MTTP and ACC mRNA abundance were negatively correlated with the level of promoter methylation. The extent of DNA methylation of the cytosine-guanine (CpG) sites in the SREBP-1c, FAS, PPARα, and LXRα promoter regions was also analyzed by direct sequencing but none differed between FLS and NFLS birds. Taken together, these results specify link DNA methylation to the pathogenesis of FLS in chickens. PMID:27083546

  18. Recombinant thermoactive phosphoenolpyruvate carboxylase (PEPC) from Thermosynechococcus elongatus and its coupling with mesophilic/thermophilic bacterial carbonic anhydrases (CAs) for the conversion of CO2 to oxaloacetate.

    Science.gov (United States)

    Del Prete, Sonia; De Luca, Viviana; Capasso, Clemente; Supuran, Claudiu T; Carginale, Vincenzo

    2016-01-15

    With the continuous increase of atmospheric CO2 in the last decades, efficient methods for carbon capture, sequestration, and utilization are urgently required. The possibility of converting CO2 into useful chemicals could be a good strategy to both decreasing the CO2 concentration and for achieving an efficient exploitation of this cheap carbon source. Recently, several single- and multi-enzyme systems for the catalytic conversion of CO2 mainly to bicarbonate have been implemented. In order to design and construct a catalytic system for the conversion of CO2 to organic molecules, we implemented an in vitro multienzyme system using mesophilic and thermophilic enzymes. The system, in fact, was constituted by a recombinant phosphoenolpyruvate carboxylase (PEPC) from the thermophilic cyanobacterium Thermosynechococcus elongatus, in combination with mesophilic/thermophilic bacterial carbonic anhydrases (CAs), for converting CO2 into oxaloacetate, a compound of potential utility in industrial processes. The catalytic procedure is in two steps: the conversion of CO2 into bicarbonate by CA, followed by the carboxylation of phosphoenolpyruvate with bicarbonate, catalyzed by PEPC, with formation of oxaloacetate (OAA). All tested CAs, belonging to α-, β-, and γ-CA classes, were able to increase OAA production compared to procedures when only PEPC was used. Interestingly, the efficiency of the CAs tested in OAA production was in good agreement with the kinetic parameters for the CO2 hydration reaction of these enzymes. This PEPC also revealed to be thermoactive and thermostable, and when coupled with the extremely thermostable CA from Sulphurhydrogenibium azorense (SazCA) the production of OAA was achieved even if the two enzymes were exposed to temperatures up to 60 °C, suggesting a possible role of the two coupled enzymes in biotechnological processes. PMID:26712095

  19. Associations of polymorphisms in the promoter I of bovine acetyl-CoA carboxylase-alpha gene with beef fatty acid composition.

    Science.gov (United States)

    Zhang, S; Knight, T J; Reecy, J M; Wheeler, T L; Shackelford, S D; Cundiff, L V; Beitz, D C

    2010-08-01

    The objectives of this study were to identify single nucleotide polymorphisms (SNPs) in the promoter I (PI) region of the bovine acetyl-CoA carboxylase-alpha (ACACA) gene and to evaluate the extent to which they were associated with lipid-related traits. Eight novel SNPs were identified, which were AJ276223:g.2064T>A (SNP1), g.2155C>T (SNP2), g.2203G>T (SNP3), g.2268T>C (SNP4), g.2274G>A (SNP5), g.2340A>G (SNP6), g.2350T>C (SNP7) and g.2370A>G (SNP8). Complete linkage disequilibrium was observed among SNP1, 2, 4, 5, 6 and 8. Phenotypic data were collected from 573 cross-bred steers with six sire breeds, including Hereford, Angus, Brangus, Beefmaster, Bonsmara and Romosinuano. The genotypes of SNP1/2/4/5/6/8 were significantly associated with adjusted backfat thickness. The genotypes of SNP3 were significantly associated with triacylglycerol (TAG) content and fatty acid composition of longissimus dorsi muscle (LM) in Brangus-, Romosinuano- and Bonsmara-sired cattle. Cattle with g.2203GG genotype had greater concentrations of TAG, total lipid, total saturated fatty acid and total monounsaturated fatty acid than did cattle with g.2203GT genotype. The genotypes of SNP7 were significantly associated with fatty acid composition of LM. Cattle with genotype g.2350TC had greater amounts of several fatty acids in LM than did cattle with genotype g.2350CC. Our results suggested that the SNPs in the PI region of ACACA gene are associated with variations in the fatty acid contents in LM. PMID:20002363

  20. Pyruvate Carboxylase Is Up-Regulated in Breast Cancer and Essential to Support Growth and Invasion of MDA-MB-231 Cells.

    Directory of Open Access Journals (Sweden)

    Phatchariya Phannasil

    Full Text Available Pyruvate carboxylase (PC is an anaplerotic enzyme that catalyzes the carboxylation of pyruvate to oxaloacetate, which is crucial for replenishing tricarboxylic acid cycle intermediates when they are used for biosynthetic purposes. We examined the expression of PC by immunohistochemistry of paraffin-embedded breast tissue sections of 57 breast cancer patients with different stages of cancer progression. PC was expressed in the cancerous areas of breast tissue at higher levels than in the non-cancerous areas. We also found statistical association between the levels of PC expression and tumor size and tumor stage (P < 0.05. The involvement of PC with these two parameters was further studied in four breast cancer cell lines with different metastatic potentials; i.e., MCF-7, SKBR3 (low metastasis, MDA-MB-435 (moderate metastasis and MDA-MB-231 (high metastasis. The abundance of both PC mRNA and protein in MDA-MB-231 and MDA-MB-435 cells was 2-3-fold higher than that in MCF-7 and SKBR3 cells. siRNA-mediated knockdown of PC expression in MDA-MB-231 and MDA-MB-435 cells resulted in a 50% reduction of cell proliferation, migration and in vitro invasion ability, under both glutamine-dependent and glutamine-depleted conditions. Overexpression of PC in MCF-7 cells resulted in a 2-fold increase in their proliferation rate, migration and invasion abilities. Taken together the above results suggest that anaplerosis via PC is important for breast cancer cells to support their growth and motility.

  1. Transit peptides of nuclear-encoded chloroplast proteins share a common amino acid framework.

    OpenAIRE

    Karlin-Neumann, G A; Tobin, E M

    1986-01-01

    We have identified three major blocks of amino acid homology shared by the transit peptides of two nuclear-encoded chloroplast proteins, the light-harvesting chlorophyll a/b-protein (LHCP) II of the thylakoid membrane and the small subunit (SSU) of ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) of the stroma. These previously unrecognized homology blocks lie at the beginning, middle and end of both transit sequences, and are separated by differing lengths of unshared (interblock) s...

  2. Characterization of acetyl-CoA and propionyl-CoA carboxylases encoded by Leptospira interrogans serovar Lai: an initial biochemical study for leptospiral gluconeogenesis via anaplerotic CO2 assimilation

    Institute of Scientific and Technical Information of China (English)

    Nanqiu Peng; Yi Zhong; Qing Zhang; Mingyue Zheng; Wei Zhao; Hualiang Jiang; Chen Yang; Xiaokui Guo; Guoping Zhao

    2012-01-01

    Leptospira interrogans is the causative agent of leptospirosis.The in vitro growth of L.interrogans requires CO2 and a partial 3-hydroxypropionate pathway involving two acyl-CoA carboxylases was suggested by genomic analysis to assimilate CO2.Either set of the candidate genes heterologously co-expressed in Escherichia coli was able to demonstrate both acetyl-CoA carboxylase (ACC)and propionyl-CoA carboxylase (PCC) activities.The trisubunit holoenzyme (LA_2736-LA_2735 and LA_3803),although failed to be purified,was designated ACC based on its substrate preference toward acetyl-CoA.The partially purified bi-subunit holoenzyme (LA_2432-LA_2433) has a considerably higher activity against propionyi-CoA as the substrate than that of acetyl-CoA,and thus,designated PCC.Native polyacrylamide gel electrophoresis indicated that this PCC has a molecular mass of around 669 kDa,suggesting an α4β4 quaternary structure and both structural homology modeling and site-directed mutagenesis analysis of its carboxyltransferase subunit (LA_2433) indicated that the A431 residue located at the bottom of the putative substrate binding pocket may play an important role in substrate specificity determination.Both transcriptomic and proteomic data indicated that enzymes involved in the suggested partial 3-hydroxypropionate pathway were expressed in vivo in addition to ACC/PCC and the homologous genes in genomes of other Leptospira species were re-annotated accordingly.However,as the in vitro detected specific activity of ACC in the crude cell extract was too low to account for the growth of the bacterium in Ellinghausen-McCulloughJohnson-Harris minimal medium,further systematic analysis is required to unveil the mechanism of gluconeogenesis via anaplerotic CO2 assimilation in Leptospira species.

  3. GenBank blastx search result: AK062203 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK062203 001-046-G09 AF497098.1 Amorphophallus smithsonianus ribulose-1,5-bisphosph...ate carboxylase/oxygenase large subunit (rbcL) gene, partial cds; chloroplast gene for chloroplast product.|PLN PLN 4e-86 +3 ...

  4. The bacterial signal transduction protein GlnB regulates the committed step in fatty acid biosynthesis by acting as a dissociable regulatory subunit of acetyl-CoA carboxylase.

    Science.gov (United States)

    Gerhardt, Edileusa C M; Rodrigues, Thiago E; Müller-Santos, Marcelo; Pedrosa, Fabio O; Souza, Emanuel M; Forchhammer, Karl; Huergo, Luciano F

    2015-03-01

    Biosynthesis of fatty acids is one of the most fundamental biochemical pathways in nature. In bacteria and plant chloroplasts, the committed and rate-limiting step in fatty acid biosynthesis is catalyzed by a multi-subunit form of the acetyl-CoA carboxylase enzyme (ACC). This enzyme carboxylates acetyl-CoA to produce malonyl-CoA, which in turn acts as the building block for fatty acid elongation. In Escherichia coli, ACC is comprised of three functional modules: the biotin carboxylase (BC), the biotin carboxyl carrier protein (BCCP) and the carboxyl transferase (CT). Previous data showed that both bacterial and plant BCCP interact with signal transduction proteins belonging to the PII family. Here we show that the GlnB paralogues of the PII proteins from E. coli and Azospirillum brasiliense, but not the GlnK paralogues, can specifically form a ternary complex with the BC-BCCP components of ACC. This interaction results in ACC inhibition by decreasing the enzyme turnover number. Both the BC-BCCP-GlnB interaction and ACC inhibition were relieved by 2-oxoglutarate and by GlnB uridylylation. We propose that the GlnB protein acts as a 2-oxoglutarate-sensitive dissociable regulatory subunit of ACC in Bacteria. PMID:25557370

  5. Small pleasures

    OpenAIRE

    Wood, Philippa

    2010-01-01

    This book simply examines the differences between what men and women consider their 'small pleasures' in life. The book combines contemporary text matter juxtaposed with traditional illustrations. A double-sided concertina book that uses the theme of minutiae as both topic and form.

  6. Study of the properties of Ribulose 1,5-biphosphate carboxylase/oxygenase from maize (Zea mays) and wheat (Triticum aestivum) by incorporation of CO2 marking 14C

    International Nuclear Information System (INIS)

    After a bibliografic review of the properties of RuBP-carboxylase/oxygenase, a methodology is described which allows the treatment of a large number of samples for the assay of the enzyme activity. 14CO3HNa is used as a marker for the counting of the incorporated radioactivity as acid insoluble material. 14''CC2 from the labeled sodium bicarbonate is the species used by the enzyme both as an activator as well as a substrate. The following experiments are described and its results given: Determination of the optimal conditions for the activation of the enzyme; study of the kinetics of the catalytic action; effect of the Mg2 concentration and determination of the Km(s) from CO2 and ribulose 1,5-biphosphate; also determination of the optimum pH at different concentrations of CO22 and Mg2. (Author) 64 refs

  7. 杜氏盐藻磷酸烯醇式丙酮酸羧化酶基因的克隆和分析%Cloning and analysis of phosphoenolpyruvate carboxylase (PEPC) gene of Dunaliella salina

    Institute of Scientific and Technical Information of China (English)

    张楠楠; 潘卫东; 崔玉琳; 秦松; 薛乐勋

    2011-01-01

    为研究杜氏盐藻(Dunaliella salina)磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PEPC)基因的功能, 根据莱茵衣藻(Chlamydomonas reinhardtii)、拟南芥(Arabidopsis thaliana)、花生(Arachis hypogaea)等真核生物PEPC 基因高度保守序列, 设计一对简并引物, 通过RT-PCR 的方法获得杜氏盐藻PEPC 基因部分序列, 然后采用RACE 的方法分别克隆到5′端和3′端序列, 拼接后得到全长cDNA, 其长度为3 523bp, 包含2 949 bp 的完整开放阅读框, 编码982 个氨基酸, 相对分子质量为110560.5。氨基酸序列与已知物种PEPC 序列的同源性依次为: Chlamydomonas reinhardtii 69%, Chlorellavariabilis 55%, Ostreococcus tauri 50%和Ostreococcus lucimarinus CCE9901 49%, 表明所克隆的序列确为杜氏盐藻PEPC cDNA 序列。%To investigate the function of phosphoenolpyruvate carboxylase (PEPC) gene of Dunaliella salina, a pair of degenerate primers was designed according to the conserved motifs of the PEPC of Chlamydomonas reinhardtii, Arabidopsis thaliana, and Arachis hypogaea. A cDNA fragment was obtained from green alga Dunaliella salina through RT-PCR, and then the full length of the cDNA was isolated by 3’ and 5’RACE. The isolated cDNA sequence was 3523 bp in length with a 2949 bp coding region that encoded 982 amino acid residues with the predicted relative molecular mass of 110560.5 dolton. In addition, homology analysis showed that PEPC of D. salina was highly similar to that of C. reinhardtii(69%), Chlorella variabilis(55%), Ostreococcus tauri(50%), and O. lucimarinus CCE9901(49%), suggesting that the cDNA isolated from Dunaliella salina was PEPC-encoding.

  8. Small Business Development Center

    Data.gov (United States)

    Small Business Administration — Small Business Development Centers (SBDCs) provide assistance to small businesses and aspiring entrepreneurs throughout the United States and its territories. SBDCs...

  9. Role of the Rubisco small subunit. Final report for period May 1, 1997--April 30,2000

    Energy Technology Data Exchange (ETDEWEB)

    Spreitzer, Robert J.

    2000-10-04

    CO{sub 2} and O{sub 2} are mutually competitive at the active site of ribulose-1,5-biphosphate (RuBP) carboxylase/oxygenase (Rubisco). Rubisco contains two subunits, each present in eight copies. The 15-kD small subunit is coded by a family of nuclear RbcS genes. Until now, the role of the small subunit in Rubisco structure or catalytic efficiency is not known. Because of other work in eliminating the two RbcS genes in the green algo Chlamydomonas reinhardtii, it is now possible to address questions about the structure-function relationships of the eukaryotic small subunit. There are three specific aims in this project: (1) Alanine scanning mutagenesis is being used to dissect the importance of the {beta}A/{beta}B loop, a feature unique to the eukaryotic small subunit. (2) Random mutagenesis is being used to identify additional residues or regions of the small subunit that are important for holoenzyme assembly and function. (3) Attempts are being made to express foreign small subunits in Chlamydomonas to examine the contribution of small subunits to holoenzyme assembly, catalytic efficiency, and CO{sub 2}/O{sub 2} specificity.

  10. Plant, cell, and molecular mechanisms of abscisic-acid regulation of stomatal apertures. In vivo phosphorylation of phosphoenolpyruvate carboxylase in guard cells of Vicia faba L. is enhanced by fusicoccin and suppressed by abscisic acid

    Energy Technology Data Exchange (ETDEWEB)

    Du, Z.; Aghoram, K.; Outlaw, W.H. Jr.

    1996-12-31

    Plants regulate water loss and CO{sub 2} gain by modulating the aperture sizes of stomata that penetrate the epidermis. Aperture size itself is increased by osmolyte accumulation and consequent turgor increase in the pair of guard cells that flank each stoma. Guard-cell phosphoenolpyruvate carboxylase, which catalyzes the regulated step leading to malate synthesis, is crucial for charge and pH maintenance during osmolyte accumulation. Regulation of this cytosolic enzyme by effectors is well documented, but additional regulation by posttranslational modification is predicted by the alteration of PEPC kinetics during stomatal opening. In this study, the authors have investigated whether this alteration is associated with the phosphorylation status of this enzyme. Using sonicated epidermal peels (isolated guard cells) pre-loaded with {sub 32}PO{sub 4}, the authors induced stomatal opening and guard-cell malate accumulation by incubation with 5 {micro}M fusicoccin (FC). In corroboratory experiments, guard cells were incubated with 5 {micro}M fusicoccin (FC). In corroboratory experiments, guard cells were incubated with the FC antagonist, 10 {micro}M abscisic acid (ABA). The phosphorylation status of PEPC was assessed by immunoprecipitation, electrophoresis, immunoblotting, and autoradiography. PEPC was phosphorylated when stomata were stimulated to open, and phosphorylation was lessened by incubation with ABA.

  11. AcEST: DK944135 [AcEST

    Lifescience Database Archive (English)

    Full Text Available y10Aa OS=Baci... 32 1.7 sp|Q9BYV9|BACH2_HUMAN Transcription regulator protei...A8QIH7_MORAL Chloroplast ribulose-1,5-bisphosphate car... 271 2e-71 tr|A9NV87|A9NV87_PICSI Putative uncharacterized pr...e-53 tr|Q06B51|Q06B51_ACERU Chloroplast ribulose-1,5-bisphosphate car... 210 5e-53 tr|A9PG25|A9PG25_POPTR Putative uncharacteri...DDGSCLY 312 >tr|Q06B48|Q06B48_ACERU Chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase activase large protein is...|Q7X9A0|RCA1_LARTR Ribulose bisphosphate carboxylase/oxygenase activase 1, chloroplastic OS=Larrea triden

  12. Lung cancer - small cell

    Science.gov (United States)

    Cancer - lung - small cell; Small cell lung cancer; SCLC ... About 15% of all lung cancer cases are SCLC. Small cell lung cancer is slightly more common in men than women. Almost all cases of SCLC ...

  13. Lung cancer - small cell

    Science.gov (United States)

    Cancer - lung - small cell; Small cell lung cancer; SCLC ... About 15% of all lung cancer cases are SCLC. Small cell lung cancer is slightly more common in men than women. Almost all cases of SCLC are ...

  14. BRANDING IN SMALL BUSINESS

    OpenAIRE

    Mihai Răzvan Constantin BARBU; Radu Florin OGARCĂ; Cătălin Mihail BARBU

    2010-01-01

    In this paper we analyzed the branding in small business. Using a desk research on Internet and the press we have identified the practices small businesses use to enhance their brand and the brand dynamics in small business. Our main contribution is that we tried to figure out the strategy of branding in small business. This need further to be investigated in order to understand how branding works in small business and to better capture the role of branding in small business.

  15. Small Business Size Standards

    Data.gov (United States)

    Small Business Administration — Certain government programs, such as SBA loan programs and contracting opportunities, are reserved for small business concerns. In order to qualify, businesses must...

  16. A comparative study of drug resistance mechanism associated with active site and non-active site mutations: I388N and D425G mutants of acetyl-coenzyme-A carboxylase.

    Science.gov (United States)

    Zhu, Xiao-Lei; Yang, Guang-Fu

    2012-03-01

    A major concern in the development of acetyl-CoA carboxylase-inhibiting (ACCase; EC 6.4.1.2) herbicides is the emergence of resistance as a result of the selection of distinct mutations within the CT domain. Mutations associated with resistance have been demonstrated to include both active sites and non-active sites, including Ile-1781-Leu, Trp- 2027-Cys, Ile-2041-Asn, Asp-2078-Gly, and Gly-2096-Ala (numbered according to the Alopecurus myosuroides plastid ACCase). In the present study, extensive computational simulations, including molecular dynamics (MD) simulations and molecular mechanics-Poisson-Boltzmann surface area (MM/PBSA) calculations, were carried out to compare the molecular mechanisms of active site mutation (I388N) and non-active site mutation (D425G) in Alopecurus myosuroides resistance to some commercial herbicides targeting ACCase, including haloxyfop (HF), diclofop (DF) and fenoxaprop (FR). All of the computational model and energetic results indicated that both I388N and D425G mutations have effects on the conformational change of the binding pocket. The π-π interaction between ligand and Phe377 and Tyr161' residues, which make an important contribution to the binding affinity, was decreased after mutation. As a result, the mutant-type ACCase has a lower affinity for the inhibitor than the wild-type enzyme, which accounts for the molecular basis of herbicidal resistance. The structural and mechanistic insights obtained from the present study will deepen our understanding of the interactions between ACCase and herbicides, which provides a molecular basis for the future design of a promising inhibitor with low resistance risk. PMID:22242795

  17. Contribution of carbon fixed by Rubisco and PEPC to phloem export in the Crassulacean acid metabolism plant Kalanchoë daigremontiana

    OpenAIRE

    Wild, Birgit; Wanek, Wolfgang; Postl, Wolfgang; Richter, Andreas

    2010-01-01

    Crassulacean acid metabolism (CAM) plants exhibit a complex interplay between CO2 fixation by phosphoenolpyruvate carboxylase (PEPC) and ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), and carbon demand for CAM maintenance and growth. This study investigated the flux of carbon from PEPC and direct Rubisco fixation to different leaf carbon pools and to phloem sap over the diurnal cycle. Concentrations and carbon isotope compositions of starch, soluble sugars, and organic acids were ...

  18. N-acetylcysteine and fructose-1,6-bisphosphate: immunomodulatory effects on mononuclear cell culture N-acetilcisteína e frutose-1,6-bisfosfato: efeito imunomodulador em cultura de células mononucleares

    Directory of Open Access Journals (Sweden)

    Ricardo Obalski de Mello

    2012-04-01

    Full Text Available INTRODUCTION: Sepsis is a complex syndrome caused by an uncontrolled systemic inflammatory response. Inflammatory cytokines play a pivotal role in septic shock pathogenesis. Therapeutic strategies have been tested in order to modulate the excessive generation or function of sepsis mediators. OBJECTIVE: The objective of the present study was to investigate the therapeutic effect of N-acetylcysteine (NAC and its association with fructose-1,6-bisphosphate (FBP on T-lymphocytes proliferation, interleukin-1β (IL-1β and monocyte chemotactic protein-1 (MCP-1 levels. MATERIAL AND METHODS: Peripheral blood mononuclear cell samples were isolated from healthy individuals. T-lymphocytes were stimulated with phytohemagglutinin for 96 hours and submitted to different concentrations of NAC or NAC associated with FBP. RESULTS: NAC (10 and 15 mM and NAC (15 mM associated with FBP reduced T-lymphocytes proliferation. IL-1β levels rose in the presence of both NAC (15 mM and NAC with FBP (1.25 mM. MCP-1 levels were reduced only by NAC (15 mM associated with FBP (1.25 mM. CONCLUSION: The results suggest that both NAC itself and NAC associated with FBP inhibit cellular proliferation, acting as potent immunomodulatory agents, which corroborates its use in the treatment of inflammatory diseases.INTRODUÇÃO: A sepse é uma síndrome complexa causada pela resposta inflamatória sistêmica descontrolada. As citocinas inflamatórias representam papel central na patogênese do choque séptico. Têm sido testadas estratégias terapêuticas a fim de modular a geração ou a função excessiva de mediadores na sepse. OBJETIVO: O objetivo deste estudo foi investigar o efeito terapêutico da N-acetilcisteína (NAC e sua associação com a frutose-1,6-bisfosfato (FBP sobre a proliferação de linfócitos T e a geração de interleucina-1β (IL-1β e proteína quimiotática de monócitos 1 (MCP-1 em cultura celular. MATERIAL E MÉTODOS: Foram isoladas células mononucleares de

  19. Small wind turbines

    CERN Document Server

    Wood, David

    2011-01-01

    Small Wind Turbines provides a thorough grounding in analysing, designing, building, and installing a small wind turbine. Small turbines are introduced by emphasising their differences from large ones and nearly all the analysis and design examples refer to small turbines.The accompanying software includes MATLAB(R) programs for power production and starting performance, as well as programs for detailed multi-objective optimisation of blade design. A spreadsheet is also given to help readers apply the simple load model of the IEC standard for small wind turbine safety. Small Wind Turbines repr

  20. BRANDING IN SMALL BUSINESS

    Directory of Open Access Journals (Sweden)

    Mihai Răzvan Constantin BARBU

    2010-01-01

    Full Text Available In this paper we analyzed the branding in small business. Using a desk research on Internet and the press we have identified the practices small businesses use to enhance their brand and the brand dynamics in small business. Our main contribution is that we tried to figure out the strategy of branding in small business. This need further to be investigated in order to understand how branding works in small business and to better capture the role of branding in small business.

  1. Uptake and expression of bacterial and cyanobacterial genes by isolated cucumber etioplasts

    International Nuclear Information System (INIS)

    The uptake and expression by plastids isolated from dark-grown cucumber cotyledons (etioplasts) of two pUC derivatives, pCS75 and pUC9-CM, respectively carrying genes for the large and small subunits of ribulose bisphosphate carboxylase/oxygenase of Anacystis nidulans or chloramphenicol acetyltransferase, is reported. Untreated etioplasts take up only 3% as much DNA as that taken up by EDTA-washed etioplasts after 2 hr of incubation with nick-translated [32P]-pCS75. The presence or absence of light does not affect DNA uptake, binding, or breakdown by etioplasts. Calcium or magnesium ions inhibit DNA uptake by 86% but enhance binding and breakdown of donor DNA by EDTA-treated etioplasts. Uncouplers that abolish membrane potential, transmembrane proton gradient, or both do not affect DNA uptake, binding, or breakdown by etioplasts. However, both DNA uptake and binding are severely inhibited by ATP. After the incubation of EDTA-treated etioplasts with pCS75, immunoprecipitation using antiserum to the small subunit of ribulose bisphosphate carboxylase/oxygenase from A. nidulans reveals the synthesis of small subunits. Treatment of etioplasts with 10 mM EDTA shows a 10-min duration to be optimal for the expression of chloramphenicol acetyltransferase encoded by pUC9-CM. A progressive increase in the expression of this enzyme is observed with an increase in the concentration of pUC9-CM in the DNA uptake medium. The plasmid-dependent incorporation of [35S] methionine by EDTA-treated organelles declines markedly during cotyledon greening in vivo

  2. Small bowel resection - discharge

    Science.gov (United States)

    ... incision is red, warm, swollen, or more painful Short of breath or chest pain Swollen legs or pain in your calves Alternative Names Small intestine surgery - discharge; Bowel resection - small intestine - discharge; Resection of ...

  3. Rapid switch of hepatic fatty acid metabolism from oxidation to esterification during diurnal feeding of meal-fed rats correlates with changes in the properties of acetyl-CoA carboxylase, but not of carnitine palmitoyltransferase I.

    Science.gov (United States)

    Moir, A M; Zammit, V A

    1993-01-01

    The effects of the ingestion of a meal on the partitioning of hepatic fatty acids between oxidation and esterification were studied in vivo for meal-fed rats. The time course for the reversal of the starved state was extremely rapid and the process was complete within 2 h, in marked contrast with the reversal of the effects of starvation in rats fed ad libitum [A. M. B. Moir and V. A. Zammit (1993) Biochem. J. 289, 49-55]. This rapid reversal occurred in spite of the fact that, in the liver of the meal-fed animals before feeding, a similar degree of partitioning of fatty acids in favour of oxidation was observed as in 24 h-starved rats (previously fed ad libitum). This suggested that the lower degree of ketonaemia observed in meal-fed rats before a meal is not due to the inability of acylcarnitine formation to compete successfully with esterification of fatty acids to the glycerol moiety. Investigation of the possible mechanisms that could contribute towards the rapid switching-off of fatty acid oxidation revealed that this was correlated with a very rapid rise and overshoot in hepatic malonyl-CoA concentration, but not with any change in the activity, or sensitivity to malonyl-CoA, of the mitochondrial overt carnitine palmitoyltransferase (CPT I). The role of these two parameters in the reversal of fasting-induced hepatic fatty acid oxidation was thus the inverse of that observed previously for refed 24 h-starved rats. The rapid increase in [malonyl-CoA] was accompanied by an immediate and complete reversion of the kinetic characteristics (Ka for citrate, expressed/total activity ratio) of acetyl-CoA carboxylase to those found in the post-meal animals, again in contrast with the time course observed in refed 24 h-starved rats [A. M. B. Moir and V. A. Zammit (1990) Biochem. J. 272, 511-517]. The rapidity with which these changes occurred was specific to the partitioning of acyl-CoA; the meal-induced diversion of glycerolipids towards phospholipid synthesis and the

  4. Securitization and small business

    OpenAIRE

    Wilcox, James A.

    2011-01-01

    Small businesses have relied considerably on securitized markets for credit. The recent financial crisis led to a virtual cessation of securitization of some of the loans used by small businesses, such as commercial real estate mortgages, vehicle, and credit card loans. In addition, values of commercial and residential real estate, which small businesses often use as collateral for loans, dropped dramatically. As a consequence, small businesses may have experienced tighter credit conditions t...

  5. Chairing a Small Department.

    Science.gov (United States)

    Bowker, Lee H.; Lynch, David M.

    Ten management problems for chairs of small departments in small colleges are discussed, along with problem-solving strategies for these administrators. Serious disagreements within a small and intimate department may create a country club culture in which differences are smoothed over and the personal idiosyncrasies of individual members are…

  6. A molecular phylogeny of the heterokont algae based on analyses of choroplast-encoded rbcL sequence data

    DEFF Research Database (Denmark)

    Daugbjerg, Niels; Andersen, Robert A.

    1997-01-01

    Nearly complete ribulose-1,5-bisphosphate carboxylase/ oxygenase (rbcL)sequences from 27 taxa of heterokont algae were determined and combined with rbcL sequences obtained from GenBank for four other heterokont algae and three red algae. The phylogeny of the morphologically diverse haterokont algae...... was inferred from an unambiguously aligned data matrix using the red algae as the root, Significantly higher levels of mutational saturation in third codon positions were found when plotting the pair-wise substitutions with and without corrections for multiple substitutions at the same site for first...... algae. In addition, each carotenoid was mapped onto both the rbcL tree and a consensus tree derived from nuclear-encoded small-subunit ribosomal DNA (SSU rDNA) sequences. Carotenoid pigmentation does not provide unambiguous phylogenetic information, whether analyzed cladistically by itself or when...

  7. AcEST: DK953168 [AcEST

    Lifescience Database Archive (English)

    Full Text Available 3168|Adiantum capillus-veneris mRNA, clone: TST39A01NGRL0017_A24, 5' (612 letters) Datab...168 Tissue type prothallia with plantlets Developmental stage gametophytes with s...9-3402. Query= DK953168|Adiantum capillus-veneris mRNA, clone: TST39A01NGRL0017_A24, 5' (612 let...) Value sp|Q8MT36|MES4_DROME Probable histone-lysine N-methyltransferase... 30 9....7 sp|P04716|RBS_SYNP6 Ribulose bisphosphate carboxylase small chai... 30 9.8 >sp|Q8MT36|MES4_DROME Probable histone-lysine N-met

  8. Influence of the nitrate concentration and source in the incorporation of 14{sub C}O2 by the RuBP-carboxylase from wheat (triticum aestivum) and maize (zea mays); Influencia de la concentracion y fuente de nitrogeno en la incorporacion de 14{sub C}O2 por la RuBp-carboxilasa de trigo (triticum aewtivum) y maiz (zea mays)

    Energy Technology Data Exchange (ETDEWEB)

    Saez Angulo, R. M.; Gines Diaz, M. J.; Garcia Pineda, M. D.

    1982-07-01

    The effect of the concentration and source of nitrogen in the culture media has been studied regarding its influence in the activity of the RuBP-carboxylase from wheat and maize during the first month of development. Wheat and maize has been chosen as plants representatives of two different types of CO{sub 2} assimilation: C3 and M- respectively. Plants have been grown in hydroponic media and under temperature, humidity and nutrient salts control. A negative effect of NH{sub 4} has been observed in the enzymatic activity of wheat seedlings, being this effect more remarkable as NH{sub 4} concentration increases and as long the time of treatment. In our experimental conditions the most favorable source of nitrogen has been N0{sub 3}NH{sub 4}. The specific activity of the enzyme from wheat is about four times higher than in maize, even it decreases with time. This decreasing has not been observed in maize, with the exception of total absence of nitrogen in the media. We have not seen significant differences between the two photo periods which have been tested. Also, no differences have been found in the enzyme activities at the different NO{sub 3}NH{sub 4} concentrations assayed, and it seems that RuBP-carboxylase metabolism is only affected in the case of absolute stress. (Author) 20 refs.

  9. Small cell glioblastoma or small cell carcinoma

    DEFF Research Database (Denmark)

    Hilbrandt, Christine; Sathyadas, Sathya; Dahlrot, Rikke H;

    2013-01-01

    was admitted to the hospital with left-sided loss of motor function. A MRI revealed a 6 cm tumor in the right temporoparietal area. The histology was consistent with both glioblastoma multiforme (GBM) and small cell lung carcinoma (SCLC) but IHC was suggestive of a SCLC metastasis. PET-CT revealed...

  10. Small Intestine Disorders

    Science.gov (United States)

    ... disease Crohn's disease Infections Intestinal cancer Intestinal obstruction Irritable bowel syndrome Ulcers, such as peptic ulcer Treatment of disorders of the small intestine depends on the cause.

  11. Discoveries in Rubisco: a historical perspective

    Science.gov (United States)

    Historic discoveries and key observations related to Rubisco (ribulose 1,5-bisphosphate carboxylase/oxygenase), from 1947 to 2006, are presented. Currently, around 200 papers describing Rubisco research are published each year and the literature contains more than 5000 manuscripts on the subject. Wh...

  12. Development of Small Company

    OpenAIRE

    Fiedler, Viktor

    2015-01-01

    The subject of bachelor thesis is analysis of possibilities and directions of the development of small company. It describes the structure of small company and the problems of its development. It analyses present economical, financial and business situation inside and outside the company. In the second part author introduces some proposals for improving several parts of company.

  13. Small States in Europe

    DEFF Research Database (Denmark)

    This book offers an accessible, coherent and informative analysis of contemporary and future foreign policy challenges facing small states in Europe.......This book offers an accessible, coherent and informative analysis of contemporary and future foreign policy challenges facing small states in Europe....

  14. Small Mammal Intrigue.

    Science.gov (United States)

    Cristol, Daniel A.

    1985-01-01

    Gives introductory information about the study of small mammals including the selection and use of harmless live-traps, handling and identification, techniques for observation and trapping in the wild, and safety measures. Suggests useful references for teachers wishing to develop a small mammal study program for their students. (JHZ)

  15. SmallSat Database

    Science.gov (United States)

    Petropulos, Dolores; Bittner, David; Murawski, Robert; Golden, Bert

    2015-01-01

    The SmallSat has an unrealized potential in both the private industry and in the federal government. Currently over 70 companies, 50 universities and 17 governmental agencies are involved in SmallSat research and development. In 1994, the U.S. Army Missile and Defense mapped the moon using smallSat imagery. Since then Smart Phones have introduced this imagery to the people of the world as diverse industries watched this trend. The deployment cost of smallSats is also greatly reduced compared to traditional satellites due to the fact that multiple units can be deployed in a single mission. Imaging payloads have become more sophisticated, smaller and lighter. In addition, the growth of small technology obtained from private industries has led to the more widespread use of smallSats. This includes greater revisit rates in imagery, significantly lower costs, the ability to update technology more frequently and the ability to decrease vulnerability of enemy attacks. The popularity of smallSats show a changing mentality in this fast paced world of tomorrow. What impact has this created on the NASA communication networks now and in future years? In this project, we are developing the SmallSat Relational Database which can support a simulation of smallSats within the NASA SCaN Compatability Environment for Networks and Integrated Communications (SCENIC) Modeling and Simulation Lab. The NASA Space Communications and Networks (SCaN) Program can use this modeling to project required network support needs in the next 10 to 15 years. The SmallSat Rational Database could model smallSats just as the other SCaN databases model the more traditional larger satellites, with a few exceptions. One being that the smallSat Database is designed to be built-to-order. The SmallSat database holds various hardware configurations that can be used to model a smallSat. It will require significant effort to develop as the research material can only be populated by hand to obtain the unique data

  16. Wind: small is beautiful

    Energy Technology Data Exchange (ETDEWEB)

    Vries, E. de

    2005-10-01

    The small wind sector (0.5-100 kW) is often overlooked but could provide decentralised energy systems. Small wind turbines have been used for homes, farms and small businesses for over 80 years (e.g. in the USA and the Netherlands), receiving a boost in the 1970s and 1980s following the 1973 oil crisis when a new generation of turbines entered the European and US markets. Bergey Windpower and Southwest Windpower from the USA are the market leaders in this sector in terms of sales volume but are still classed as medium-sized enterprises. Small turbines have the disadvantage of higher costs compared with large turbines due to higher manufacturing costs, technical factors associated with the tendency to use small turbines on relatively short towers, small production runs and a failure to keep up with the latest design developments such as cost-effective state-of-the-art frequency converters. Most small turbines are horizontal axis turbines, though vertical axis turbines are produced by some manufacturers. Examples of the systems available from European suppliers are described.

  17. Wind: small is beautiful

    International Nuclear Information System (INIS)

    The small wind sector (0.5-100 kW) is often overlooked but could provide decentralised energy systems. Small wind turbines have been used for homes, farms and small businesses for over 80 years (e.g. in the USA and the Netherlands), receiving a boost in the 1970s and 1980s following the 1973 oil crisis when a new generation of turbines entered the European and US markets. Bergey Windpower and Southwest Windpower from the USA are the market leaders in this sector in terms of sales volume but are still classed as medium-sized enterprises. Small turbines have the disadvantage of higher costs compared with large turbines due to higher manufacturing costs, technical factors associated with the tendency to use small turbines on relatively short towers, small production runs and a failure to keep up with the latest design developments such as cost-effective state-of-the-art frequency converters. Most small turbines are horizontal axis turbines, though vertical axis turbines are produced by some manufacturers. Examples of the systems available from European suppliers are described

  18. Small scale optics

    CERN Document Server

    Yupapin, Preecha

    2013-01-01

    The behavior of light in small scale optics or nano/micro optical devices has shown promising results, which can be used for basic and applied research, especially in nanoelectronics. Small Scale Optics presents the use of optical nonlinear behaviors for spins, antennae, and whispering gallery modes within micro/nano devices and circuits, which can be used in many applications. This book proposes a new design for a small scale optical device-a microring resonator device. Most chapters are based on the proposed device, which uses a configuration know as a PANDA ring resonator. Analytical and nu

  19. AcEST: DK949080 [AcEST

    Lifescience Database Archive (English)

    Full Text Available otein endopeptidase O... 32 2.8 sp|Q9BYV9|BACH2_HUMAN Transcription regulator protein BACH2 OS=H... 32 2.8 sp...-86 tr|Q06B49|Q06B49_ACERU Chloroplast ribulose-1,5-bisphosphate car... 317 4e-85 tr|A9PG25|A9PG25_POPTR Putative uncharacterized pr...Q06B50|Q06B50_ACERU Chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase activase large protein is...2 Definition Adiantum capillus-veneris mRNA. clone: TST38A01NGRL0005_A14. 5' end sequence. Accession DK949080 Tissue type pr...|Q7X999|RCA2_LARTR Ribulose bisphosphate carboxylase/oxygenase activase 2, chloroplastic OS=Larrea tri

  20. Small satellites - An overview

    Science.gov (United States)

    Ward, A. K.

    The present review of small satellites examines spacecraft activities in the U.K. and compiles a checklist of advantages and applications for the class. These advantages are illustrated with references to recent small satellite missions and technologies developed to facilitate such launches and projects. Specific programs examined include AMPTE-UKS, Viking, and the UoSAT program, and information is given regarding the Small Explorer program, the RAE Space Technology Research Vehicle, the AEA Argos Program, and space research programs in both Japan and India. Low-cost launches are shown to be available in the form of the Ariane Structure for Auxiliary Payloads, the Pegasus and Delta vehicles, and with the Shuttle Free-flying Getaway Special. Small-satellite technologies that play key roles in their effective implementation are: structure/thermal advances, attitude control systems, on-board communications, and power and data-handling systems.

  1. Small bowel bacterial overgrowth

    Science.gov (United States)

    ... Surgical procedures that create a loop of small intestine where excess bacteria can grow. An example is a Billroth II type of stomach removal ( gastrectomy ). Some cases of irritable bowel syndrome (IBS) Symptoms The most common symptoms are: Abdominal ...

  2. Small & Large Intestine

    Science.gov (United States)

    ... the large intestine produces no digestive enzymes. Chemical digestion is completed in the small intestine before the chyme reaches the large intestine. Functions of the large intestine include the absorption of water and electrolytes and the elimination of ...

  3. Small bowel resection

    Science.gov (United States)

    ... reconnect, your surgeon makes an opening called a stoma through the skin of your belly. The small ... of your belly. Stool will go through the stoma into a drainage bag outside your body. This ...

  4. Small - Display Cartography

    DEFF Research Database (Denmark)

    Nissen, Flemming; Hvas, Anders; Münster-Swendsen, Jørgen;

    This report comprises the work carried out in the work-package of small display cartography. The work-package has aimed at creating a general framework for the small-display cartography. A solid framework facilitates an increased use of spatial data in mobile devices - thus enabling, together with...... the rapidly evolving positioning techniques, a new category of position-dependent, map-based services to be introduced. The report consists of the following parts: Part I: Categorization of handheld devices, Part II: Cartographic design for small-display devices, Part III: Study on the GiMoDig Client...... ? Portal Service Communication and finally, Part IV: Concluding remarks and topics for further research on small-display cartography. Part II includes a separate Appendix D consisting of a cartographic design specification. Part III includes a separate Appendix C consisting of a schema specification, a...

  5. Small Satellite Transporter Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The primary objective is to determine whether this small satellite transporter is capable of transporting at least four 6U CubeSats is possible for a given set of...

  6. Small Mammal Trapping 2003

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Small mammal traps were placed in the Baring division and in the Edmunds division of Moosehom National Wildlife Refuge. There were a total of 98 traps set for up to...

  7. Research using small tokamaks

    International Nuclear Information System (INIS)

    These proceedings of the IAEA-sponsored meeting held in Nice, France 10-11 October, 1988, contain the manuscripts of the 21 reports dealing with research using small tokamaks. The purpose of this meeting was to highlight some of the achievements of small tokamaks and alternative magnetic confinement concepts and assess the suitability of starting new programs, particularly in developing countries. Papers presented were either review papers, or were detailed descriptions of particular experiments or concepts. Refs, figs and tabs

  8. SMALL SCALE MORPHODYNAMICAL MODELLING

    Institute of Scientific and Technical Information of China (English)

    D. Ditschke; O. Gothel; H. Weilbeer

    2001-01-01

    Long term morphological simulations using complete coupled models lead to very time consuming computations. Latteux (1995) presented modelling techniques developed for tidal current situations in order to reduce the computational effort. In this paper the applicability of such methods to small scale problems is investigated. It is pointed out that these methods can be transferred to small scale problems using the periodicity of the vortex shedding process.

  9. Small-x physics

    Energy Technology Data Exchange (ETDEWEB)

    Mueller, A.H. [Columbia Univ., New York, NY (United States). Dept. of Physics

    1997-06-01

    After a brief review of the kinematics of deep inelastic lepton-proton scattering, the parton model is described. Small-x behavior coming from DGLAP evolution and from BFKL evolution is discussed, and the two types of evolution are contrasted and compared. Then a more detailed discussion of BFKL dynamics is given. The phenomenology of small-x physics is discussed with an emphasis on ways in which BFKL dynamics may be discussed and measured. 45 refs., 12 figs.

  10. UNDERSTANDING SMALL BUSINESS SCAMS

    OpenAIRE

    Michael T. Schaper; PAUL WEBER

    2012-01-01

    This paper provides an overview of the current state of knowledge about small business scams. A scam is a form of dishonest action, based upon an invitation to participate in an activity. Victims are encouraged, mislead or induced to voluntarily interact with the perpetrator, and ultimately to willingly surrender over money, information or other valuable resources. Common forms of scams directed towards small business include phishing, false business valuations and sales, fake overpayments, f...

  11. Radionuclide Small Intestine Imaging

    OpenAIRE

    Jiri Dolezal; Marcela Kopacova

    2013-01-01

    The aim of this overview article is to present the current possibilities of radionuclide scintigraphic small intestine imaging. Nuclear medicine has a few methods—scintigraphy with red blood cells labelled by means of 99mTc for detection of the source of bleeding in the small intestine, Meckel's diverticulum scintigraphy for detection of the ectopic gastric mucosa, radionuclide somatostatin receptor imaging for carcinoid, and radionuclide inflammation imaging. Video capsule or deep enteroscop...

  12. Small test SDHW systems

    DEFF Research Database (Denmark)

    Vejen, Niels Kristian

    1999-01-01

    Three small test SDHW systems was tested in a laboratory test facility.The three SDHW systems where all based on the low flow principe and a mantle tank but the design of the systems where different.......Three small test SDHW systems was tested in a laboratory test facility.The three SDHW systems where all based on the low flow principe and a mantle tank but the design of the systems where different....

  13. Small simple impact craters

    CERN Document Server

    Sparavigna, Amelia Carolina

    2010-01-01

    The paper discusses some examples of image processing applied to improve optical satellite imagery of small craters (Kamil, Veevers, Haviland). The examples show that image processing can be quite useful for further in-situ researches, because the resultant imagery helps to have a better picture of the crater shape and of the distribution of debris about it. The paper is also disclosing an interesting underwater structure, with shape and size of a small crater, located on the coast-line of Sudan.

  14. Small wind turbine

    OpenAIRE

    Vélez Castellano, Didier

    2010-01-01

    The main objective is to develop a project on installing a small wind turbine at the University of Glyndwr in Wrexham Wales. Today are immersed in a world seeking clean energy for reduce greenhouse gases because this problem is becoming a global reality. So installing a small wind turbine at the university would provide large quantity of clean energy to supply a workshop and also reduce the expulsion of CO2 into the atmosphere. The main characteristic of the turbine under...

  15. Gas Quenching Small Components

    Institute of Scientific and Technical Information of China (English)

    Paul Stratton

    2004-01-01

    Gas quenching is an environmentally friendly and effective way of quenching components. However it has not been possible to apply it to small components because of the difficulties of jigging. A method of gas quenching such small components is described in which they are levitated in a confined gas stream. The method is suitable for quenching low alloy steel components that weigh between 1 and 10g.

  16. How small is a small cloud?

    Directory of Open Access Journals (Sweden)

    I. Koren

    2008-07-01

    Full Text Available The interplay between clouds and aerosols and their contribution to the radiation budget is one of the largest uncertainties of climate change. Most work to date has separated cloudy and cloud-free areas in order to evaluate the individual radiative forcing of aerosols, clouds, and aerosol effects on clouds.

    Here we examine the size distribution and the optical properties of small, sparse cumulus clouds and the associated optical properties of what is considered a cloud-free atmosphere within the cloud field. We show that any separation between clouds and cloud free atmosphere will incur errors in the calculated radiative forcing.

    The nature of small cumulus cloud size distributions suggests that at any resolution, a significant fraction of the clouds are missed, and their optical properties are relegated to the apparent cloud-free optical properties. At the same time, the cloudy portion incorporates significant contribution from non-cloudy pixels.

    We show that the largest contribution to the total cloud reflectance comes from the smallest clouds and that the spatial resolution changes the apparent energy flux of a broken cloudy scene. When changing the resolution from 30 m to 1 km (Landsat to MODIS the average "cloud-free" reflectance at 1.65 μm increases from 0.0095 to 0.0115 (>20%, the cloud reflectance decreases from 0.13 to 0.066 (~50%, and the cloud coverage doubles, resulting in an important impact on climate forcing estimations. The apparent aerosol forcing is on the order of 0.5 to 1 Wm−2 per cloud field.

  17. Beech trees exposed to high CO{sub 2} and to simulated summer ozone levels: Effects on photosynthesis, chloroplast components and leaf enzyme activity

    Energy Technology Data Exchange (ETDEWEB)

    Lutz, C. [Univ. of Innsbruck, Inst. of Botany, Innsbruck (Austria); Anegg, S. [GFS. National Research Centre for Environment and Health, Inst. of Biochemical Plant Pathology, Oberschleissheim (Germany); Gerant, D.; Dizengremel, P. [Univ. Henri Poincare Nancy 1, Lab. de Biologie Forestiere, Vandauvre les Nancy cedex (France); Alaoui-Sosse, B. [Lab. de Biologie et Ecophysiologie, Besancon cedex (France)

    2000-07-01

    Young trees of European beech (Fagus sylvatica L.) were exposed in a phytotron to different levels of ozone and CO{sub 2} under the climatic simulation of one vegetation period. High ozone levels were simulated similar to high ozone concentration in the field (up to 110 ppb), while CO{sub 2} was added as 300 ppm to the present level of ca 380 ppm. Our study describes different aspects of photosynthesis from the leaf level to the reactions of selected thylakoid components at different harvest times during growth of the beech trees under the different fumigation regimes. Ozone effects appeared in the first weeks of the treatment as a stimulation of chlorophyll fluorescence (F{sub v}/F{sub m}), in oxygen production and in ribulose-1,5-bisphosphate carboxylase/oxygenase activity, while the summer and early autumn harvests showed strong reductions in these parameters. Only phosphoenolpyruvate carboxylase (PEPcase) activity remained higher under high ozone. The effects of high CO{sub 2} appeared in general as a small stimulation in enzyme activity like PEPcase in spring. However, with increasing time of fumigation, reductions of all parameters were observed. Especially chlorophylls showed strong reductions under high CO{sub 2}. The combined treatment with high ozone plus high CO{sub 2} resulted mostly in an amelioration of the negative ozone effects, although control levels were not reached. (au)

  18. Small Intestinal Infections.

    Science.gov (United States)

    Munot, Khushboo; Kotler, Donald P

    2016-06-01

    Small intestinal infections are extremely common worldwide. They may be bacterial, viral, or parasitic in etiology. Most are foodborne or waterborne, with specific etiologies differing by region and with diverse pathophysiologies. Very young, very old, and immune-deficient individuals are the most vulnerable to morbidity or mortality from small intestinal infections. There have been significant advances in diagnostic sophistication with the development and early application of molecular diagnostic assays, though these tests have not become mainstream. The lack of rapid diagnoses combined with the self-limited nature of small intestinal infections has hampered the development of specific and effective treatments other than oral rehydration. Antibiotics are not indicated in the absence of an etiologic diagnosis, and not at all in the case of some infections. PMID:27168147

  19. Small Animal Retinal Imaging

    Science.gov (United States)

    Choi, WooJhon; Drexler, Wolfgang; Fujimoto, James G.

    Developing and validating new techniques and methods for small animal imaging is an important research area because there are many small animal models of retinal diseases such as diabetic retinopathy, age-related macular degeneration, and glaucoma [1-6]. Because the retina is a multilayered structure with distinct abnormalities occurring in different intraretinal layers at different stages of disease progression, there is a need for imaging techniques that enable visualization of these layers individually at different time points. Although postmortem histology and ultrastructural analysis can be performed for investigating microscopic changes in the retina in small animal models, this requires sacrificing animals, which makes repeated assessment of the same animal at different time points impossible and increases the number of animals required. Furthermore, some retinal processes such as neurovascular coupling cannot be fully characterized postmortem.

  20. Small hydroelectric engineering practice

    CERN Document Server

    Leyland, Bryan

    2014-01-01

    Small Hydroelectric Engineering Practice is a comprehensive reference book covering all aspects of identifying, building, and operating hydroelectric schemes between 500 kW and 50 MW. In this range of outputs there are many options for all aspects of the scheme and it is very important that the best options are chosen.As small hydroelectric schemes are usually built against a limited budget it is extremely important that the concept design is optimum and every component is designed to maximise the benefi t and minimise the cost. As operating costs are often a high proportion of the income it i

  1. Small is beautiful

    Energy Technology Data Exchange (ETDEWEB)

    Woof, M.; Casteel, K.

    1997-12-01

    While safety standards in modern mechanised underground coal mines in Britain are good, so are standards in their smaller competitors using much older equipment. Working practices and conditions at several Federation of Independent Mines (FIM) operations are described, detailing the use of equipment and methods that would be considered obsolete in larger mines. Although these mines are small the high quality of the coal that they mine means that they are economic. Drilling equipment used in small mines, and scoops are also described. 7 photos. 1 tab.

  2. How Small are Small Stars Really?

    Science.gov (United States)

    2002-11-01

    VLT Interferometer Measures the Size of Proxima Centauri and Other Nearby Stars [1] Summary At a distance of only 4.2 light-years, Proxima Centauri is the nearest star to the Sun currently known [2]. It is visible as an 11-magnitude object in the southern constellation of Centaurus and is the faintest member of a triple system, together with Alpha Centauri , the brightest (double) star in this constellation. Proxima Centauri is a very-low-mass star, in fact barely massive enough to burn hydrogen to helium in its interior. It is about seven times smaller than the Sun, and the surface temperature is "only" about 3000 degrees, about half of that of our own star. Consequently, it is also much fainter - the intrinsic brightness is only 1/150th of that of our Sun. Low-mass stars are very interesting objects , also because the physical conditions in their interiors have much in common with those of giant planets, like Jupiter in our solar system. A determination of the sizes of the smallest stars has been impossible until now because of their general faintness and lack of adequate instrumentation. However, astronomers have long been keen to move forward in this direction, since such measurements would provide indirect, crucial information about the behaviour of matter under extreme conditions. When the first observations with the VLT Interferometer (VLTI), combining the light from two of the 8.2-m VLT Unit Telescopes (ANTU and MELIPAL), were made one year ago ( ESO PR 23/01 ), interferometric measurements were also obtained of Proxima Centauri . They formed part of the VLTI commissioning and the data were soon released to the ESO community, cf. the special website. Now, an international team of astronomers from Switzerland, France and ESO/Chile has successfully analysed these observations by means of newly developed, advanced software. For the first time ever, they obtained a highly accurate measurement of the size of such a small star . Three other small stars were also

  3. Small public private partnerships

    DEFF Research Database (Denmark)

    Koch, Christian; Jensen, Jesper Ole

    2009-01-01

    authorities, which recommend sizes over 13,5 million € (100 million DKr). PPP is here understood as Design, Build, Finance and Operate projects. The paper shows, when looking at Germany, Italy and United Kingdom, that small PPP (below 13,5 million €) are widespread in two investigated countries; United...

  4. Small hydrogen liquefier

    International Nuclear Information System (INIS)

    In this work the deign and construction of a small hydrogen liquefier (two liters per hour maximum production) is described. The isenthalpic expansion process is used, because its construction is simple and it is generally cheaper to operate. A comparison with other liquefier processes, and considerations about their basic theory are also presented. (author)

  5. Small Public Library Management

    Science.gov (United States)

    Pearlmutter, Jane; Nelson, Paul

    2012-01-01

    Anyone at the helm of a small public library knows that every little detail counts. But juggling the responsibilities that are part and parcel of the job is far from easy. Finally, here's a handbook that includes everything administrators need to keep a handle on library operations, freeing them up to streamline and improve how the organization…

  6. Imaging the small bowel.

    LENUS (Irish Health Repository)

    Murphy, Kevin P

    2014-03-01

    Radiologic investigations continue to play a pivotal role in the diagnosis of pathologic conditions of the small intestine despite enhancement of capsule endoscopy and double-balloon endoscopy. Imaging techniques continue to evolve and new techniques in MRI in particular, are being developed.

  7. Small Group Research

    Science.gov (United States)

    McGrath, Joseph E.

    1978-01-01

    Summarizes research on small group processes by giving a comprehensive account of the types of variables primarily studied in the laboratory. These include group structure, group composition, group size, and group relations. Considers effects of power, leadership, conformity to social norms, and role relationships. (Author/AV)

  8. Small can be beautiful

    Energy Technology Data Exchange (ETDEWEB)

    Albrand, Solveig (Institut des Sciences Nucleaires, IN2P3-CNRS/Universite Joseph Fourier, 53, avenue des Martyrs, 38026, Grenoble Cedex (France)); Ravel, J.C. (Institut des Sciences Nucleaires, IN2P3-CNRS/Universite Joseph Fourier, 53, avenue des Martyrs, 38026, Grenoble Cedex (France))

    1994-12-15

    Small laboratories with limited financial means can use low-cost personal computers for many control applications. Examples of control systems developed at SARA are discussed, illustrating the evolution of the system in step with that of the PC, but each time with a considerable re-use of software. ((orig.))

  9. Small is beautiful

    Energy Technology Data Exchange (ETDEWEB)

    Hindley, M.

    1994-09-01

    This article focusses on small hydroelectric power development in Finland. Legislative and financial incentives, and use of standard tubular turbines are discussed. The Vuotos reservoir project is described, and the preparation of an environmental assessment of the project is noted. (UK)

  10. Research using small tokamaks

    International Nuclear Information System (INIS)

    This document consists of a collection of papers presented at the IAEA Technical Committee Meeting on Research Using Small Tokamaks. It contains 22 papers on a wide variety of research aspects, including diagnostics, design, transport, equilibrium, stability, and confinement. Some of these papers are devoted to other concepts (stellarators, compact tori). Refs, figs and tabs

  11. Small and Magical

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    Ant genome study offers promising evidence in medical and anti-aging research While children often love observing ants happily for hours, grownups are more likely to ignore these small creatures. Some scientists, however, show greater interest in ants than children do.

  12. Small is Beatiful

    DEFF Research Database (Denmark)

    Hansen, Henrik Aldershaab Boje; Pleman, Allan; Koch, Christian

    1999-01-01

    The life and death of a small Danish software house developing ERP-software in collaboration with costumers is analysed.User producer relationships are central drivers in the company's history. This reactive development strategy creates a niche at the marketplace but produces internal...

  13. Small Windmills in Denmark

    DEFF Research Database (Denmark)

    Petersen, H.

    1980-01-01

    The report describes a project for small windmills funded by the Ministry of Energy. The test plant is described and a survey of Danish windmills is presented. Some requirements for windmills are mentioned and regulations governing the interface between grid-connected windmills and the electric...

  14. Deburring small intersecting holes

    Energy Technology Data Exchange (ETDEWEB)

    Gillespie, L.K.

    1980-08-01

    Deburring intersecting holes is one of the most difficult deburring tasks faced by many industries. Only 14 of the 37 major deburring processes are applicable to most intersecting hole applications. Only five of these are normally applicable to small or miniature holes. Basic process capabilities and techniques used as a function of hole sizes and intersection depths are summarized.

  15. Libraries for Small Museums.

    Science.gov (United States)

    Anderson, Linda M.

    Presented are the very basic requirements for establishing a small special library operating under a limited budget. Physical plant organization, cataloging, book processing, circulation procedures, book selection and ordering and instructions for typists are covered. Although the practices discussed were established for a museum library, what is…

  16. Small hydroelectric power plants

    International Nuclear Information System (INIS)

    Small hydroelectric power plants are power plants of 1 - 10 MW. For a supplier, this is an unnatural limit. A more natural limit involves compact engine design and simplified control system. The article discusses most of the engine and electrotechnical aspects in the development, construction and operation of such a plant

  17. 77 FR 30227 - Small Business Size Regulations, Small Business Innovation Research (SBIR) Program and Small...

    Science.gov (United States)

    2012-05-22

    ... BUSINESS ADMINISTRATION 13 CFR Part 121 Small Business Size Regulations, Small Business Innovation Research (SBIR) Program and Small Business Technology Transfer (STTR) Program AGENCY: U.S. Small Business... to amend its regulations governing size and eligibility for the Small Business Innovation...

  18. Small Engine & Accessory Test Area

    Data.gov (United States)

    Federal Laboratory Consortium — The Small Engine and Accessories Test Area (SEATA) facilitates testaircraft starting and auxiliary power systems, small engines and accessories. The SEATA consists...

  19. [Small intestine bacterial overgrowth].

    Science.gov (United States)

    Leung Ki, E L; Roduit, J; Delarive, J; Guyot, J; Michetti, P; Dorta, G

    2010-01-27

    Small intestine bacterial overgrowth (SIBO) is a condition characterised by nutrient malabsorption and excessive bacteria in the small intestine. It typically presents with diarrhea, flatulence and a syndrome of malabsorption (steatorrhea, macrocytic anemia). However, it may be asymptomatic in the eldery. A high index of suspicion is necessary in order to differentiate SIBO from other similar presenting disorders such as coeliac disease, lactose intolerance or the irritable bowel syndrome. A search for predisposing factor is thus necessary. These factors may be anatomical (stenosis, blind loop), or functional (intestinal hypomotility, achlorydria). The hydrogen breath test is the most frequently used diagnostic test although it lacks standardisation. The treatment of SIBO consists of eliminating predisposing factors and broad-spectrum antibiotic therapy. PMID:20214190

  20. Small steps for hydro

    International Nuclear Information System (INIS)

    The government in Peru has decided to utilise its gas reserves and restrict hydro to relatively small schemes. A number of reasons for the decision are given. In 1997, the Shell-Mobile-Bechtel-COSAPI consortium was formed and agreements were signed regarding exploiting Gas de Camisea. The country's energy needs to 2010 are being assessed. It is likely that by 2001 the whole of south Peru will be receiving gas from Camisea. The Peru situation is discussed under the headings of (i) existing capacity, (ii) growing demands, (iii) a history of hydro in Peru, (iv) electrification and SHP and (v) outlook. The future for Peru's electric energy development is bright. While most of its new power capacity will come from natural gas, the small hydros also have a part to play. A stronger commitment of national and regional political authorities to consider supplies outside the big cities is said to be needed. (UK)

  1. Small Wind Energy Systems

    DEFF Research Database (Denmark)

    Simoes, Marcelo; Farret, Felix Alberto; Blaabjerg, Frede

    2015-01-01

    devices, and a centralized distribution control. In order to establish a small wind energy system it is important to observe the following: (i) Attending the energy requirements of the actual or future consumers; (ii) Establishing civil liabilities in case of accidents and financial losses due to shortage...... costs and guaranteeing an acceptable energy supply. This paper discuss how performance is affected by local conditions and random nature of the wind, power demand profiles, turbine related factors, and presents the technical issues for implementing a self-excited induction generator system, or a......The application of small wind turbines for residential and commercial applications depends on how a microgrid can operate in a suitable way. Because there is variable demand and a random nature of wind resources, it is usually necessary the installation of controllable end-use loads, storage...

  2. Advanced Small Rechargeable Batteries

    Science.gov (United States)

    Halpert, Gerald

    1989-01-01

    Lithium-based units offer highest performance. Paper reviews status of advanced, small rechargeable batteries. Covers aqueous systems including lead/lead dioxide, cadmium/nickel oxide, hydrogen/nickel oxide, and zinc/nickel oxide, as well as nonaqueous systems. All based on lithium anodes, nonaqueous systems include solid-cathode cells (lithium/molybdenum disulfide, lithium/titanium disulfide, and lithium/vanadium oxide); liquid-cathode cells (lithium/sulfur dioxide cells); and new category, lithium/polymer cells.

  3. Small reactor return

    International Nuclear Information System (INIS)

    Current state of the development of present-day small reactors in different countries is performed. Various designs of low and middle power reactors, among which are CAREM (25 MW, PWR), KLT-40 (40 MW, PWR), MRX (30 MW, PWR), IRIS (50 MW, PWR), SMART (1000 MW, PWR), Modular SBWR (50 MW, BWR), PBMR (120 MW, HTGR), GT-HMR (285 MW, HTGR), are discussed

  4. Small and Magical

    Institute of Scientific and Technical Information of China (English)

    TANG YUANKAI

    2011-01-01

    While children often love observing ants happily for hours,grown-ups are more likely to ignore these small creatures.Some scientists,however,show greater interest in ants than children do.One of them is Zhang Guojie,Associate Director of the Bioinformatics Center and Leader of Genome Projects in BGI.BG1 is China's largest genome and bioinformatics analysis center in Shenzhen,in south China's Guangdong Province.

  5. Small Business Innovations (Photodetector)

    Science.gov (United States)

    1991-01-01

    Epitaxx, Inc. of Princeton, NJ, developed the Epitaxx Near Infrared Room Temperature Indium-Gallium-Arsenide (InGaAs) Photodetector based on their Goddard Space Flight Center Small Business Innovation Research (SBIR) contract work to develop a linear detector array for satellite imaging applications using InGaAs alloys that didn't need to be cooled to (difficult and expensive) cryogenic temperatures. The photodetectors can be used for remote sensing, fiber optic and laser position-sensing applications.

  6. Small Business Innovations (MISER)

    Science.gov (United States)

    1991-01-01

    Lightwave Electronics Corporation, Mountain View, CA, developed the Series 120 and 122 non-planner diode pumped ring lasers based on a low noise ring laser with voltage tuning that they delivered to Jet Propulsion Laboratory under a Small Business Innovation Research (SBIR) contract. The voltage tuning feature allows "phase-locking" the lasers, making them "electronic," similar to radio and microwave electronic oscillators. The Series 120 and 122 can be applied to fiber sensing, coherent communications and laser radar.

  7. Leadership in Small Societies

    OpenAIRE

    Stephen Younger

    2010-01-01

    Multi-agent simulation was used to study several styles of leadership in small societies. Populations of 50 and100 agents inhabited a bounded landscape containing a fixed number of food sources. Agents moved about the landscape in search of food, mated, produced offspring, and died either of hunger or at a predetermined maximum age. Leadership models focused on the collection and redistribution of food. The simulations suggest that individual households were more effective at meeting their ne...

  8. Think Small Go Big

    Institute of Scientific and Technical Information of China (English)

    汤维维

    2006-01-01

    Vepoo公司在创立之前,经历了三次创业转型。用他们的话来说,从“think big go small”转到“think small go big”用了一年的时间。这期间他们耗尽了初期筹备资金,幸运的是在最后一刻迎来了黎明的曙光。

  9. Small Cell City

    OpenAIRE

    Dehghan, S.; Steele, R.

    1997-01-01

    Traditionally, mobile operators have planned their networks to accommodate mobile terminals at ground level. Increasingly, mobile users communicate while stationary from within high-rise buildings. With mobiles operating at a variety of different heights and mobilities, plus the requirement to accommodate increasing teletraffic and multimedia services, there is a need to compact small cells into the three-dimensional city space. This article is concerned with using city buildings to act as el...

  10. Small Animal Bone Biomechanics

    OpenAIRE

    Vashishth, Deepak

    2008-01-01

    Animal models, in particular mice, offer the possibility of naturally achieving or genetically engineering a skeletal phenotype associated with disease and conducting destructive fracture tests on bone to determine the resulting change in bone’s mechanical properties. Several recent developments, including nano- and micro- indentation testing, microtensile and microcompressive testing, and bending tests on notched whole bone specimens, offer the possibility to mechanically probe small animal ...

  11. When small is beautiful. Boom time for small hydro?

    Energy Technology Data Exchange (ETDEWEB)

    Osborne, John

    2002-08-01

    Although small hydropower projects generally contribute a small proportion of most nations' electricity needs their importance often exceeds their size. The author, a freelance industrial journalist considers prospects for small hydro, how such schemes are benefiting communities throughout the world and why some countries are more successfully using small hydro than others. (Author)

  12. The small library manager's handbook

    CERN Document Server

    Graves, Alice

    2014-01-01

    The Small Library Manager's Handbook is for librarians working in all types of small libraries. It covers the everyday nuts-and-bolts operations that all librarians must perform. This handbook, written by experts who are small librarians themselves, will help all small librarians to do multiple jobs at the same time.

  13. Spacecraft at Small NEO

    CERN Document Server

    Scheeres, D

    2006-01-01

    The impact risk has been sharply curtailed for the largest km-sized Near Earth Objects (NEOs) through a concerted period of observation spanning the last decade. Thus the risk of impact has now migrated to the smaller and more numerous members of the Near Earth Object (NEO) population. Characterization and mitigation missions to these smaller objects become more complex from a spacecraft operations perspective, even as the prospects for nudging such lower mass bodies off course become more feasible. This paper details the challenges for spacecraft operations at small bodies and indicates areas where more research and analysis of this problem is needed.

  14. Rapid small lot manufacturing

    Energy Technology Data Exchange (ETDEWEB)

    Harrigan, R.W.

    1998-05-09

    The direct connection of information, captured in forms such as CAD databases, to the factory floor is enabling a revolution in manufacturing. Rapid response to very dynamic market conditions is becoming the norm rather than the exception. In order to provide economical rapid fabrication of small numbers of variable products, one must design with manufacturing constraints in mind. In addition, flexible manufacturing systems must be programmed automatically to reduce the time for product change over in the factory and eliminate human errors. Sensor based machine control is needed to adapt idealized, model based machine programs to uncontrolled variables such as the condition of raw materials and fabrication tolerances.

  15. Rolling at small scales

    DEFF Research Database (Denmark)

    Nielsen, Kim L.; Niordson, Christian F.; Hutchinson, John W.

    2016-01-01

    The rolling process is widely used in the metal forming industry and has been so for many years. However, the process has attracted renewed interest as it recently has been adapted to very small scales where conventional plasticity theory cannot accurately predict the material response. It is well...... plasticity. Metals are known to be stronger when large strain gradients appear over a few microns; hence, the forces involved in the rolling process are expected to increase relatively at these smaller scales. In the present numerical analysis, a steady-state modeling technique that enables convergence...

  16. Small changes for works

    Institute of Scientific and Technical Information of China (English)

    Jiarui Wu

    2011-01-01

    The central dogma of molecular biology indicates that genetic information from DNA is transmitted to RNA,and then translated into proteins that play various roles during all physiological and pathological processes.However,most of the proteins have to make small chemical changes on their amino acid chains,so-called post-translational modifications,before they start to work.The collection of four articles in this issue presents timely developments in the area of protein modifications,ranging from how proteins carry out their biological functions based on protein modifications to what are molecular mechanisms of protein modifications.

  17. Business Small company

    OpenAIRE

    KREJČOVÁ, Kateřina

    2011-01-01

    This diploma work deals with a Business Small company. The work moves on the concrete company, which is analyzed and on the basis of analyses the new strategy is formulated. At the theoretical part, summary of solving problems is introduced. It’s needed for the analytic part of work. At the beginning, business is defined in economic terms. Found in this part are basic concepts that are linked to business. From the general definition of business the work fluently proceeds to more concrete...

  18. Small Business Innovations (Cryostat)

    Science.gov (United States)

    1991-01-01

    General Pneumatics Corporation, Scottsdale, AZ, developed an anti- clogging cryostat that liquifies gases by expansion for high pressure through a nozzle to produce cryorefrigeration based on their Kennedy Space Center Small Business Innovation Research (SBIR) work to develop a Joule-Thomson (JT) expansion valve that is less susceptible to clogging by particles or condensed contaminants in the flow than a non-contaminating compressor in a closed cycle Linde-Hampson cryocooler used to generate cryogenic cooling for infrared sensors, super conductors, supercooled electronics and cryosurgery.

  19. Small Business Innovations (Helicopters)

    Science.gov (United States)

    1992-01-01

    The amount of engine power required for a helicopter to hover is an important, but difficult, consideration in helicopter design. The EHPIC program model produces converged, freely distorted wake geometries that generate accurate analysis of wake-induced downwash, allowing good predictions of rotor thrust and power requirements. Continuum Dynamics, Inc., the Small Business Innovation Research (SBIR) company that developed EHPIC, also produces RotorCRAFT, a program for analysis of aerodynamic loading of helicopter blades in forward flight. Both helicopter codes have been licensed to commercial manufacturers.

  20. Small mirror fusion reactors

    International Nuclear Information System (INIS)

    Basic requirements for the pilot plants are that they produce a net product and that they have a potential for commercial upgrade. We have investigated a small standard mirror fusion-fission hybrid, a two-component tandem mirror hybrid, and two versions of a field-reversed mirror fusion reactor--one a steady state, single cell reactor with a neutral beam-sustained plasma, the other a moving ring field-reversed mirror where the plasma passes through a reaction chamber with no energy addition

  1. Novel small-molecule AMPK activator orally exerts beneficial effects on diabetic db/db mice

    Energy Technology Data Exchange (ETDEWEB)

    Li, Yuan-Yuan; Yu, Li-Fang; Zhang, Li-Na; Qiu, Bei-Ying; Su, Ming-Bo; Wu, Fang; Chen, Da-Kai; Pang, Tao; Gu, Min; Zhang, Wei; Ma, Wei-Ping; Jiang, Hao-Wen; Li, Jing-Ya, E-mail: jyli@mail.shcnc.ac.cn; Nan, Fa-Jun, E-mail: fjnan@mail.shcnc.ac.cn; Li, Jia, E-mail: jli@mail.shcnc.ac.cn

    2013-12-01

    AMP-activated protein kinase (AMPK), which is a pivotal guardian of whole-body energy metabolism, has become an attractive therapeutic target for metabolic syndrome. Previously, using a homogeneous scintillation proximity assay, we identified the small-molecule AMPK activator C24 from an optimization based on the original allosteric activator PT1. In this paper, the AMPK activation mechanism of C24 and its potential beneficial effects on glucose and lipid metabolism on db/db mice were investigated. C24 allosterically stimulated inactive AMPK α subunit truncations and activated AMPK heterotrimers by antagonizing autoinhibition. In primary hepatocytes, C24 increased the phosphorylation of AMPK downstream target acetyl-CoA carboxylase dose-dependently without changing intracellular AMP/ATP ratio, indicating its allosteric activation in cells. Through activating AMPK, C24 decreased glucose output by down-regulating mRNA levels of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) in primary hepatocytes. C24 also decreased the triglyceride and cholesterol contents in HepG2 cells. Due to its improved bioavailability, chronic oral treatment with multiple doses of C24 significantly reduced blood glucose and lipid levels in plasma, and improved the glucose tolerance of diabetic db/db mice. The hepatic transcriptional levels of PEPCK and G6Pase were reduced. These results demonstrate that this orally effective activator of AMPK represents a novel approach to the treatment of metabolic syndrome. - Highlights: • C24 activates AMPK through antagonizing autoinhibition within α subunit. • C24 activates AMPK in hepatocytes and decreases glucose output via AMPK. • C24 exerts beneficial effects on diabetic db/db mice. • C24 represents a novel therapeutic for treatment of metabolic syndrome.

  2. Novel small-molecule AMPK activator orally exerts beneficial effects on diabetic db/db mice

    International Nuclear Information System (INIS)

    AMP-activated protein kinase (AMPK), which is a pivotal guardian of whole-body energy metabolism, has become an attractive therapeutic target for metabolic syndrome. Previously, using a homogeneous scintillation proximity assay, we identified the small-molecule AMPK activator C24 from an optimization based on the original allosteric activator PT1. In this paper, the AMPK activation mechanism of C24 and its potential beneficial effects on glucose and lipid metabolism on db/db mice were investigated. C24 allosterically stimulated inactive AMPK α subunit truncations and activated AMPK heterotrimers by antagonizing autoinhibition. In primary hepatocytes, C24 increased the phosphorylation of AMPK downstream target acetyl-CoA carboxylase dose-dependently without changing intracellular AMP/ATP ratio, indicating its allosteric activation in cells. Through activating AMPK, C24 decreased glucose output by down-regulating mRNA levels of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) in primary hepatocytes. C24 also decreased the triglyceride and cholesterol contents in HepG2 cells. Due to its improved bioavailability, chronic oral treatment with multiple doses of C24 significantly reduced blood glucose and lipid levels in plasma, and improved the glucose tolerance of diabetic db/db mice. The hepatic transcriptional levels of PEPCK and G6Pase were reduced. These results demonstrate that this orally effective activator of AMPK represents a novel approach to the treatment of metabolic syndrome. - Highlights: • C24 activates AMPK through antagonizing autoinhibition within α subunit. • C24 activates AMPK in hepatocytes and decreases glucose output via AMPK. • C24 exerts beneficial effects on diabetic db/db mice. • C24 represents a novel therapeutic for treatment of metabolic syndrome

  3. Securitization of Small Business Loans

    OpenAIRE

    Beshouri, Christopher P.; Peter J. Nigro

    1995-01-01

    This paper assesses the potential impact of securitization in improving small businesses’ access to credit. It begins by examining the nature of small business lending and the factors that make banks the primary providers of credit to small businesses. The paper then examines the conditions under which the benefits of securitization are fully realized and whether the nature of small business lend­ing satisfies those conditions. We argue that certain characteristics of small firm finance, espe...

  4. Measurement of 2-carboxyarabinitol 1-phosphate in plant leaves by isotope dilution

    International Nuclear Information System (INIS)

    The level of 2-carboxyarabinitol 1-phosphate (CA1P) in leaves of 12 species was determined by an isotope dilution assay. 14C-labeled standard was synthesized from [2-14C]carboxyarabinitol 1,5-bisphosphate using acid phosphatase, and was added at the initial point of leaf extraction. Leaf CA1P was purified and its specific activity determined. CA1P was found in dark-treated leaves of all species examined, including spinach (Spinacea oleracea), wheat (Triticum aestivum), Arabidopsis thaliana, and maize (Zea mays). The highest amounts were found in bean (Phaseolus vulgaris) and petunia (Petunia hybrida), which had 1.5 to 1.8 moles CA1P per mole ribulose 1,5-bisphosphate carboxylase catalytic sites. Most species had intermediate amounts of CA1P (0.2 to 0.8 mole CA1P per mole catalytic sites). Such intermediate to high levels of CA1P support the hypothesis that CA1P functions in many species as a light-dependent regulator of ribulose 1,5-bisphosphate carboxylase activity and whole leaf photosynthetic CO2 assimilation. However, CA1P levels in spinach, wheat, and A. thaliana were particularly low (less than 0.09 mole CA1P per mole catalytic sites). In such species, CA1P does not likely have a significant role in regulating ribulose 1,5-bisphosphate carboxylase activity, but could have a different physiological role

  5. 77 FR 76215 - Small Business Size Regulations, Small Business Innovation Research (SBIR) Program and Small...

    Science.gov (United States)

    2012-12-27

    ... (available at http://www.gpo.gov/fdsys/pkg/FR-2012-05-15/pdf/2012-11586.pdf ), the U.S. Small Business... ADMINISTRATION 13 CFR Part 121 RIN 3245-AG46 Small Business Size Regulations, Small Business Innovation Research... regulations governing size and eligibility for the Small Business Innovation Research (SBIR) and...

  6. 78 FR 11745 - Small Business Size Regulations, Small Business Innovation Research (SBIR) Program and Small...

    Science.gov (United States)

    2013-02-20

    ... RIN 3245-AG46 Small Business Size Regulations, Small Business Innovation Research (SBIR) Program and... related to size and eligibility for the Small Business Innovation Research (SBIR) and Small Business... . SUPPLEMENTARY INFORMATION: On May 15, 2012, at 77 FR 28520 (available at...

  7. 77 FR 28520 - Small Business Size Regulations, Small Business Innovation Research (SBIR) Program and Small...

    Science.gov (United States)

    2012-05-15

    ... ADMINISTRATION 13 CFR Part 121 RIN 3245-AG46 Small Business Size Regulations, Small Business Innovation Research... its regulations governing size and eligibility for the Small Business Innovation Research (SBIR) and.... Background On July 22, 1982, Congress enacted and the President signed into law the Small Business...

  8. Small caliber guided projectile

    Science.gov (United States)

    Jones, James F.; Kast, Brian A.; Kniskern, Marc W.; Rose, Scott E.; Rohrer, Brandon R.; Woods, James W.; Greene, Ronald W.

    2010-08-24

    A non-spinning projectile that is self-guided to a laser designated target and is configured to be fired from a small caliber smooth bore gun barrel has an optical sensor mounted in the nose of the projectile, a counterbalancing mass portion near the fore end of the projectile and a hollow tapered body mounted aft of the counterbalancing mass. Stabilizing strakes are mounted to and extend outward from the tapered body with control fins located at the aft end of the strakes. Guidance and control electronics and electromagnetic actuators for operating the control fins are located within the tapered body section. Output from the optical sensor is processed by the guidance and control electronics to produce command signals for the electromagnetic actuators. A guidance control algorithm incorporating non-proportional, "bang-bang" control is used to steer the projectile to the target.

  9. Small intestinal transplantation.

    LENUS (Irish Health Repository)

    Quigley, E M

    2012-02-03

    The past few years have witnessed a considerable shift in the clinical status of intestinal transplantation. A great deal of experience has been gained at the most active centers, and results comparable with those reported at a similar stage in the development of other solid-organ graft programs are now being achieved by these highly proficient transplant teams. Rejection and its inevitable associate, sepsis, remain ubiquitous, and new immunosuppressant regimes are urgently needed; some may already be on the near horizon. The recent success of isolated intestinal grafts, together with the mortality and morbidity attendant upon the development of advanced liver disease related to total parenteral nutrition, has prompted the bold proposal that patients at risk for this complication should be identified and should receive isolated small bowel grafts before the onset of end-stage hepatic failure. The very fact that such a suggestion has begun to emerge reflects real progress in this challenging field.

  10. Small Business Innovations

    Science.gov (United States)

    1996-01-01

    Under a Small Business Innovation Research (SBIR) contract to Kennedy Space Center, EIC Laboratories invented a Raman Spectrograph with fiber optic sampling for space applications such as sensing hazardous fuel vapors and making on-board rapid analyses of chemicals and minerals. Raman spectroscopy is a laser-based measurement technique that provides through a unique vibrational spectrum a molecular 'fingerprint,' and can function in aqueous environments. EIC combined optical fiber technology with Raman methods to develop sensors that can be operated at a distance from the spectrographic analysis instruments and the laser excitation source. EIC refined and commercialized the technology to create the Fiber Optic Raman Spectrograph and the RamanProbe. Commercial applications range from process control to monitoring hazardous materials.

  11. Seminar on small PWRs

    International Nuclear Information System (INIS)

    The first of the three papers presented, reviews the history of small and medium power reactors. Although the category includes all reactors with an output up to 600MWe, the concentration is on the range 250-300MWe. Their suitability for developing countries and their economic aspects are discussed. The other two papers are concerned with the construction, transportation, installation and management of the Shore Test Facility for the prototype of the PWR 2, a reactor designed for use in submarines. The test facility was built at Barrow-in-Furness and then moved to its site in Caithness, Scotland. The methods used to move the enormous loads involved are described in detail. The construction and installation was planned over 5 years and involved a large number of people from the construction company, the Royal Navy and the contractors who operate the facility. The project management is described. (U.K.)

  12. Small-crack test methods

    Science.gov (United States)

    Larsen, James M.; Allison, John E.

    This book contains chapters on fracture mechanics parameters for small fatigue cracks, monitoring small-crack growth by the replication method, measurement of small cracks by photomicroscopy (experiments and analysis), and experimental mechanics of microcracks. Other topics discussed are the real-time measurement of small-crack-opening behavior using an interferometric strain/displacement gage; direct current electrical potential measurement of the growth of small cracks; an ultrasonic method for the measurement of the size and opening behavior of small fatigue cracks; and the simulation of short crack and other low closure loading conditions, utilizing constant K(max) Delta-K-decreasing fatigue crack growth procedures.

  13. Small Active Radiation Monitor

    Science.gov (United States)

    Badhwar, Gautam D.

    2004-01-01

    A device, named small active radiation monitor, allows on-orbit evaluations during periods of increased radiation, after extravehicular activities, or at predesignated times for crews on such long-duration space missions as on the International Space Station. It also permits direct evaluation of biological doses, a task now performed using a combination of measurements and potentially inaccurate simulations. Indeed the new monitor can measure a full array of radiation levels, from soft x-rays to hard galactic cosmic-ray particles. With refinement, it will benefit commercial (nuclear power-plant workers, airline pilots, medical technicians, physicians/dentists, and others) and military personnel as well as the astronauts for whom thermoluminescent dosimeters are inadequate. Civilian and military personnel have long since graduated from film badges to thermoluminescent dosimeters. Once used, most dosimeters must be returned to a central facility for processing, a step that can take days or even weeks. While this suffices for radiation workers for whom exposure levels are typically very low and of brief duration, it does not work for astronauts. Even in emergencies and using express mail, the results can often be delayed by as much as 24 hours. Electronic dosimeters, which are the size of electronic oral thermometers, and tattlers, small electronic dosimeters that sound an alarm when the dose/dose rate exceeds preset values, are also used but suffer disadvantages similar to those of thermoluminescent dosimeters. None of these devices fully answers the need of rapid monitoring during the space missions. Instead, radiation is monitored by passive detectors, which are read out after the missions. Unfortunately, these detectors measure only the absorbed dose and not the biologically relevant dose equivalent. The new monitor provides a real-time readout, a time history of radiation exposures (both absorbed dose and biologically relevant dose equivalent), and a count of the

  14. Small intestine aspirate and culture

    Science.gov (United States)

    ... ency/article/003731.htm Small intestine aspirate and culture To use the sharing features on this page, please enable JavaScript. Small intestine aspirate and culture is a lab test to check for infection ...

  15. Small intestine contrast injection (image)

    Science.gov (United States)

    ... and throat, through the stomach into the small intestine. When in place, contrast dye is introduced and ... means of demonstrating whether or not the small intestine is normal when abnormality is suspected.

  16. Brighter for small power plants

    International Nuclear Information System (INIS)

    The article presents a small tunnel drilling machine aimed at using for the construction of small hydroelectric power plants and mentions briefly some advantages economically and environmentally of both the machine and the power production solution

  17. Small Area Fair Market Rent

    Data.gov (United States)

    Department of Housing and Urban Development — Due to the increasing demand for more localized rents for a variety of purposes, HUD is making Small Area FMRs for all metropolitan areas available. Small Area FMRs...

  18. Thinking small is big again

    Energy Technology Data Exchange (ETDEWEB)

    Butler, C.; Didsbury, R.; Strati, G.L.; Sur, B. [Atomic Energy of Canada Limited, Chalk River, Ontario (Canada)

    2012-12-15

    This article introduces a series of articles in the field of small reactors. Activity in this field has been on the upswing in North America. The major advantages of a small modular reactor are improved safety, lower cost and a small-risk venture. Canada has a long history of small reactor designs for research reactors as well as power reactors. Some of these reactors are ZEEP, ZED-2, NRX, NRU, WR-1, SLOWPOKE and CANDU power reactors.

  19. Globalization, Entrepreneurship and Small Business

    OpenAIRE

    Stan ªtefania Anca

    2012-01-01

    New product innovation often comes from small business. Most the radical inventions of the last century, such as the computer, come from small companies, not large ones. And this will not change. Globalization will continue to modify and mold the environment or small business and entrepreneurship. The competition from abroad places strong pressure on small business in terms of efficiency, price, quality, cost, control, marketing, expertise, customer satisfaction and innovativeness. Globalizat...

  20. Training in Chinese Small Firms

    OpenAIRE

    Dai, Zhejun

    2007-01-01

    The dissertation aims to review the currently predominant literature on the factors influencing the training in small business in the industrialized Western economies and to carry out empirical investigation on the determinants of training in Chinese small business and then analyze whether the Western-based literature is consistent with the practices of training in Chinese small firms. In addition, relevant problems with training in small business are presented and recommendations on how to r...

  1. Small Hydropower - The comeback of small hydropower stations

    International Nuclear Information System (INIS)

    This issue of the 'Erneuerbare Energien' (renewable energies) magazine published by the Swiss Solar Energy Society takes a look at small hydropower projects in Switzerland. In a number of interviews and articles, various topics concerning small hydropower are dealt with. First of all, an interview with Bruno Guggisberg, previously responsible for small hydro at the Swiss Federal Office of Energy, examines the potential of small hydro and the various political, technical and economic influences on such projects. Further articles provide an overview of the various types of small hydro schemes, including power generation using height differences in drinking-water and wastewater installations. As far as the components of small hydro schemes are concerned, various types of turbines and further system components that are needed are examined. A further article takes a look at the small hydro market and the market players involved. Ecological aspects and research activities are discussed in further articles. In a second interview with Martin Boelli, presently responsible for small hydropower at the Swiss Federal Office of Energy, the unused potential for the use of hydropower in Switzerland is discussed. Examples of small-scale hydro schemes are examined and the support offered by the Small Hydropower Program is discussed. Finally the question is asked, if the small hydro market in Switzerland is overheated as a result of promotion schemes such as cost-covering remuneration for electricity from renewable energy sources.

  2. Modified Small Business Network Security

    Directory of Open Access Journals (Sweden)

    Md. Belayet Ali

    2012-02-01

    Full Text Available This paper covers some likely threats and effectivesteps for a secure small business. It also involves a flowchart tocomprehend the overall small business network security easilyand we identify a set of security issues and applyappropriate techniques to satisfy the correspondingsecurity requirements. In respect of all, this document isstrong enough for any small business network security.

  3. Small intestine aspirate and culture

    Science.gov (United States)

    Small intestine aspirate and culture is a lab test to check for infection in the small intestine. ... A sample of fluid from the small intestine is needed. A procedure ... done to get the sample. The fluid is placed in a special dish in ...

  4. Research using small tokamaks

    International Nuclear Information System (INIS)

    The technical reports contained in this collection of papers on research using small tokamaks fall into four main categories, i.e., (i) experimental work (heating, stability, plasma radial profiles, fluctuations and transport, confinement, ultra-low-q tokamaks, wall physics, a.o.), (ii) diagnostics (beam probes, laser scattering, X-ray tomography, laser interferometry, electron-cyclotron absorption and emission systems), (iii) theory (strong turbulence, effects of heating on stability, plasma beta limits, wave absorption, macrostability, low-q tokamak configurations and bootstrap currents, turbulent heating, stability of vortex flows, nonlinear islands growth, plasma-drift-induced anomalous transport, ergodic divertor design, a.o.), and (iv) new technical facilities (varistors applied to establish constant current and loop voltage in HT-6M), lower-hybrid-current-drive systems for HT-6B and HT-6M, radio-frequency systems for HT-6M ICR heating experimentation, and applications of fiber optics for visible and vacuum ultraviolet radiation detection as applied to tokamaks and reversed-field pinches. A total number of 51 papers are included in the collection. Refs, figs and tabs

  5. Small Hydropower in Nigeria

    Institute of Scientific and Technical Information of China (English)

    P.EOKRANEFE; S.OWOLABI

    2002-01-01

    Nigeria is the most populaous black nation in the world. It has an area of 913,072 square kilometres. The country population is about 120 million. There are three major languages, Hausa, Yoruba and Igbo, although there is still more than 380 dialects in local languages. There are large number of household on the African continent that are unelectrified.In Nigeria about 70 million people remain literally in the dark without access to electricity. The majority of these numbers are in the rural areas. This workshop is apt in a number of ways. It is a joint effort between government, private sector, the academic and other practitioners in small hydro power station, it is also a promotion of business and industrial activities as well as development of renewable energy resources. I wish to commend the organizers of this workshop. The focus of the workshop is relevant to the agenda of the Federal Ministry of Power and Steel and aspirations of the Federal Government of Nigeria to provide regular and steady electricity to majority of Nigerians before the end of year 2001.

  6. Exposition regarding small windmills

    International Nuclear Information System (INIS)

    Emphasis is laid on the elucidation of the current situation of small windmills in Denmark by estimating the industry's strengths and weaknesses and evaluating the technical status and user economy relevant to these wind turbines, in addition to their technical and marketing potentials within the next 3-5 years. It is also attempted to survey and forecast their situation within a competitive market which includes marketing and sales potential outside Denmark, often by way of development projects financed by DANIDA (Danish International Development Agency, a department under the Danish Foreign Ministry that takes responsibility for Danish aid to developing countries, often by direct agreement). It is noted that it is difficult to make predictions in the light of currently rapidly changing global political aspects, but it is suggested that sales of smaller Danish windmills within the country could amount to 300-700 wind turbines by the year 2000. Regarding sales abroad, it is concluded that the current number will not increase unless great efforts are made within the export market and the instigation of close cooperation with foreign companies which use an international sales network and have access to local distribution channels. (AB) (15 refs.)

  7. 48 CFR 970.1907 - Subcontracting with Small Business, Small Disadvantaged Business and Woman-Owned Small Business...

    Science.gov (United States)

    2010-10-01

    ... MANAGEMENT AND OPERATING CONTRACTS Small, Small Disadvantaged and Women-Owned Small Business Concerns 970.1907 Subcontracting with Small Business, Small Disadvantaged Business and Woman-Owned Small Business... Business, Small Disadvantaged Business and Woman-Owned Small Business Concerns. 970.1907 Section...

  8. Joint research using small tokamaks

    International Nuclear Information System (INIS)

    Small tokamaks have an important role in fusion research. More than 40 small tokamaks are operational. Research on small tokamaks has created a scientific basis for the scaling-up to larger tokamaks. Well-known scientific and engineering schools, which are now determining the main directions of fusion science and technology, have been established through research on small tokamaks. Combined efforts within a network of small and medium size tokamaks will further enhance the contribution of small tokamaks. A new concept of interactive coordinated research using small tokamaks in the mainstream fusion science areas, in testing of new diagnostics, materials and technologies as well as in education, training and broadening of the geography of fusion research in the scope of the IAEA Coordinated Research Project, is presented

  9. Joint research using small tokamaks

    International Nuclear Information System (INIS)

    Small tokamaks have an important role in fusion research. More than 40 small tokamaks are operational. Research on small tokamaks has created a scientific basis for the scaling-up to larger tokamaks. Well-known scientific and engineering schools, which are now determining the main directions of fusion science and technology, have been established through research on small tokamaks. Combined efforts within a network of small and medium size tokamaks will further enhance the contribution of small tokamaks. A new concept of interactive co-ordinated research using small tokamaks in the mainstream fusion science areas, in testing of new diagnostics, materials and technologies as well as in education, training and broadening of the geography of fusion research in the scope of the IAEA Co-ordinated Research Project is presented. (author)

  10. Small diameter carbon nanopipettes

    Science.gov (United States)

    Singhal, Riju; Bhattacharyya, Sayan; Orynbayeva, Zulfiya; Vitol, Elina; Friedman, Gary; Gogotsi, Yury

    2010-01-01

    Nanoscale multifunctional carbon probes facilitate cellular studies due to their small size, which makes it possible to interrogate organelles within living cells in a minimally invasive fashion. However, connecting nanotubes to macroscopic devices and constructing an integrated system for the purpose of fluid and electrical signal transfer is challenging, as is often the case with nanoscale components. We describe a non-catalytic chemical vapor deposition based method for batch fabrication of integrated multifunctional carbon nanopipettes (CNPs) with tip diameters much smaller (10-30 nm) than previously reported (200 nm and above) and approaching those observed for multiwalled carbon nanotubes. This eliminates the need for complicated attachment/assembly of nanotubes into nanofluidic devices. Variable tip geometries and structures were obtained by controlled deposition of carbon inside and outside quartz pipettes. We have shown that the capillary length and gas flow rate have a marked effect on the carbon deposition. This gives us a flexible protocol, useful for growing carbon layers of different thicknesses at selective locations on a glass pipette to yield a large variety of cellular probes in bulk quantities. The CNPs possess an open channel for fluid transfer with the carbon deposited inside at 875 °C behaving like an amorphous semiconductor. Vacuum annealing of the CNP tips at temperatures up to 2000 °C yields graphitic carbon structures with an increase in conductivity of two orders of magnitude. Penetration of the integrated carbon nanoprobes into cells was shown to produce minimal Ca2+ signals, fast recovery of basal Ca2+ levels and no adverse activation of the cellular metabolism during interrogation times as long as 0.5-1 h.

  11. Small diameter carbon nanopipettes

    International Nuclear Information System (INIS)

    Nanoscale multifunctional carbon probes facilitate cellular studies due to their small size, which makes it possible to interrogate organelles within living cells in a minimally invasive fashion. However, connecting nanotubes to macroscopic devices and constructing an integrated system for the purpose of fluid and electrical signal transfer is challenging, as is often the case with nanoscale components. We describe a non-catalytic chemical vapor deposition based method for batch fabrication of integrated multifunctional carbon nanopipettes (CNPs) with tip diameters much smaller (10-30 nm) than previously reported (200 nm and above) and approaching those observed for multiwalled carbon nanotubes. This eliminates the need for complicated attachment/assembly of nanotubes into nanofluidic devices. Variable tip geometries and structures were obtained by controlled deposition of carbon inside and outside quartz pipettes. We have shown that the capillary length and gas flow rate have a marked effect on the carbon deposition. This gives us a flexible protocol, useful for growing carbon layers of different thicknesses at selective locations on a glass pipette to yield a large variety of cellular probes in bulk quantities. The CNPs possess an open channel for fluid transfer with the carbon deposited inside at 875 deg. C behaving like an amorphous semiconductor. Vacuum annealing of the CNP tips at temperatures up to 2000 deg. C yields graphitic carbon structures with an increase in conductivity of two orders of magnitude. Penetration of the integrated carbon nanoprobes into cells was shown to produce minimal Ca2+ signals, fast recovery of basal Ca2+ levels and no adverse activation of the cellular metabolism during interrogation times as long as 0.5-1 h.

  12. Small Mercury Relativity Orbiter

    Science.gov (United States)

    Bender, Peter L.; Vincent, Mark A.

    1989-01-01

    The accuracy of solar system tests of gravitational theory could be very much improved by range and Doppler measurements to a Small Mercury Relativity Orbiter. A nearly circular orbit at roughly 2400 km altitude is assumed in order to minimize problems with orbit determination and thermal radiation from the surface. The spacecraft is spin-stabilized and has a 30 cm diameter de-spun antenna. With K-band and X-band ranging systems using a 50 MHz offset sidetone at K-band, a range accuracy of 3 cm appears to be realistically achievable. The estimated spacecraft mass is 50 kg. A consider-covariance analysis was performed to determine how well the Earth-Mercury distance as a function of time could be determined with such a Relativity Orbiter. The minimum data set is assumed to be 40 independent 8-hour arcs of tracking data at selected times during a two year period. The gravity field of Mercury up through degree and order 10 is solved for, along with the initial conditions for each arc and the Earth-Mercury distance at the center of each arc. The considered parameters include the gravity field parameters of degree 11 and 12 plus the tracking station coordinates, the tropospheric delay, and two parameters in a crude radiation pressure model. The conclusion is that the Earth-Mercury distance can be determined to 6 cm accuracy or better. From a modified worst-case analysis, this would lead to roughly 2 orders of magnitude improvement in the knowledge of the precession of perihelion, the relativistic time delay, and the possible change in the gravitational constant with time.

  13. Small angle neutron scattering

    Directory of Open Access Journals (Sweden)

    Cousin Fabrice

    2015-01-01

    Full Text Available Small Angle Neutron Scattering (SANS is a technique that enables to probe the 3-D structure of materials on a typical size range lying from ∼ 1 nm up to ∼ a few 100 nm, the obtained information being statistically averaged on a sample whose volume is ∼ 1 cm3. This very rich technique enables to make a full structural characterization of a given object of nanometric dimensions (radius of gyration, shape, volume or mass, fractal dimension, specific area… through the determination of the form factor as well as the determination of the way objects are organized within in a continuous media, and therefore to describe interactions between them, through the determination of the structure factor. The specific properties of neutrons (possibility of tuning the scattering intensity by using the isotopic substitution, sensitivity to magnetism, negligible absorption, low energy of the incident neutrons make it particularly interesting in the fields of soft matter, biophysics, magnetic materials and metallurgy. In particular, the contrast variation methods allow to extract some informations that cannot be obtained by any other experimental techniques. This course is divided in two parts. The first one is devoted to the description of the principle of SANS: basics (formalism, coherent scattering/incoherent scattering, notion of elementary scatterer, form factor analysis (I(q→0, Guinier regime, intermediate regime, Porod regime, polydisperse system, structure factor analysis (2nd Virial coefficient, integral equations, characterization of aggregates, and contrast variation methods (how to create contrast in an homogeneous system, matching in ternary systems, extrapolation to zero concentration, Zero Averaged Contrast. It is illustrated by some representative examples. The second one describes the experimental aspects of SANS to guide user in its future experiments: description of SANS spectrometer, resolution of the spectrometer, optimization of

  14. Small angle neutron scattering

    International Nuclear Information System (INIS)

    Small Angle Neutron Scattering (SANS) is a technique that enables to probe the 3-D structure of materials on a typical size range lying from about 1 nm up to a few 100 nm, the obtained information being statistically averaged on a sample whose volume is ∼ 1 cm3. This very rich technique enables to make a full structural characterization of a given object of nano-metric dimensions (radius of gyration, shape, volume or mass, fractal dimension, specific area... ) through the determination of the form factor as well as the determination of the way objects are organized within in a continuous media, and therefore to describe interactions between them, through the determination of the structure factor. The specific properties of neutrons (possibility of tuning the scattering intensity by using the isotopic substitution, sensitivity to magnetism, negligible absorption, low energy of the incident neutrons) make it particularly interesting in the fields of soft matter, biophysics, magnetic materials and metallurgy. In particular, the contrast variation methods allow to extract some information that cannot be obtained by any other experimental techniques. This course is divided in two parts. The first one is devoted to the description of the principle of SANS: basics (formalism, coherent scattering/incoherent scattering, notion of elementary scatterer), form factor analysis (I(q→0), Guinier regime, intermediate regime, Porod regime, polydisperse system), structure factor analysis (2. Virial coefficient, integral equations, characterization of aggregates), and contrast variation methods (how to create contrast in an homogeneous system, matching in ternary systems, extrapolation to zero concentration, Zero Averaged Contrast). It is illustrated by some representative examples. The second one describes the experimental aspects of SANS to guide user in its future experiments: description of SANS spectrometer, resolution of the spectrometer, optimization of spectrometer

  15. Research using small tokamaks

    International Nuclear Information System (INIS)

    The technical reports in this document were presented at the IAEA Technical Committee Meeting ''Research on Small Tokamaks'', September 1990, in three sessions, viz., (1) Plasma Modes, Control, and Internal Phenomena, (2) Edge Phenomena, and (3) Advanced Configurations and New Facilities. In Section (1) experiments at controlling low mode number modes, feedback control using external coils, lower-hybrid current drive for the stabilization of sawtooth activity and continuous (1,1) mode, and unmodulated and fast modulated ECRH mode stabilization experiments were reported, as well as the relation to disruptions and transport of low m,n modes and magnetic island growth; static magnetic perturbations by helical windings causing mode locking and sawtooth suppression; island widths and frequency of the m=2 tearing mode; ultra-fast cooling due to pellet injection; and, finally, some papers on advanced diagnostics, i.e., lithium-beam activated charge-exchange spectroscopy, and detection through laser scattering of discrete Alfven waves. In Section (2), experimental edge physics results from a number of machines were presented (positive biasing on HYBTOK II enhancing the radial electric field and improving confinement; lower hybrid current drive on CASTOR improving global particle confinement, good current drive efficiency in HT-6B showing stabilization of sawteeth and Mirnov oscillations), as well as diagnostic developments (multi-chord time resolved soft and ultra-soft X-ray plasma radiation detection on MT-1; measurements on electron capture cross sections in multi-charged ion-atom collisions; development of a diagnostic neutral beam on Phaedrus-T). Theoretical papers discussed the influence of sheared flow and/or active feedback on edge microstability, large edge electric fields, and two-fluid modelling of non-ambipolar scrape-off layers. Section (3) contained (i) a proposal to construct a spherical tokamak ''Proto-Eta'', (ii) an analysis of ultra-low-q and runaway

  16. Compound 13, an α1-selective small molecule activator of AMPK, inhibits Helicobacter pylori-induced oxidative stresses and gastric epithelial cell apoptosis.

    Science.gov (United States)

    Zhao, Hangyong; Zhu, Huanghuang; Lin, Zhou; Lin, Gang; Lv, Guoqiang

    2015-08-01

    Half of the world's population experiences Helicobacter pylori (H. pylori) infection, which is a main cause of gastritis, duodenal and gastric ulcer, and gastric cancers. In the current study, we investigated the potential role of compound 13 (C13), a novel α1-selective small molecule activator of AMP-activated protein kinase (AMPK), against H. pylori-induced cytotoxicity in cultured gastric epithelial cells (GECs). We found that C13 induced significant AMPK activation, evidenced by phosphorylation of AMPKα1 and ACC (acetyl-CoA carboxylase), in both primary and transformed GECs. Treatment of C13 inhibited H. pylori-induced GEC apoptosis. AMPK activation was required for C13-mediated GEC protection. Inhibition of AMPK kinase activity by the AMPK inhibitor Compound C, or silencing AMPKα1 expression by targeted-shRNAs, alleviated C13-induced GEC protective activities against H. pylori. Significantly, C13 inhibited H. pylori-induced reactive oxygen species (ROS) production in GECs. C13 induced AMPK-dependent expression of anti-oxidant gene heme oxygenase (HO-1) in GECs. Zinc protoporphyrin (ZnPP) and tin protoporphyrin (SnPP), two HO-1 inhibitors, not only suppressed C13-mediated ROS scavenging activity, but also alleviated its activity in GECs against H. pylori. Together, these results indicate that C13 inhibits H. pylori-induced ROS production and GEC apoptosis through activating AMPK-HO-1 signaling. PMID:26022128

  17. Photorespiration and carbon concentrating mechanisms: two adaptations to high O2, low CO2 conditions.

    Science.gov (United States)

    Moroney, James V; Jungnick, Nadine; Dimario, Robert J; Longstreth, David J

    2013-11-01

    This review presents an overview of the two ways that cyanobacteria, algae, and plants have adapted to high O2 and low CO2 concentrations in the environment. First, the process of photorespiration enables photosynthetic organisms to recycle phosphoglycolate formed by the oxygenase reaction catalyzed by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Second, there are a number of carbon concentrating mechanisms that increase the CO2 concentration around Rubisco which increases the carboxylase reaction enhancing CO2 fixation. This review also presents possibilities for the beneficial modification of these processes with the goal of improving future crop yields. PMID:23771683

  18. Successful budgeting for small business

    OpenAIRE

    Cheng, Yafang

    2006-01-01

    The most important rule for financial management in small companies, although it is general rule for all sizes of company, is not to run out of cash. Being a financial manager for a small company, it is more serious to manage company's budget because the smaller enterprise faces greater challenge in raising cash. The following subjects are areas to be covered in addressing the issue of budgeting in a small business: -the current academic views and literature reviews on the subject o...

  19. Sustainability for a Small Country

    OpenAIRE

    Edwards, Geoff W.

    1992-01-01

    The main objective in this paper is to promote thinking on what "sustainable development" or "ecologically sustainable development" means for a small trading country. After a brief acknowledgement of some of the diverse literature on sustainability, there is a consideration of some realities facing a small trading economy. The balance of the paper comprises a discussion of the scope for a small country to enhance sustainability. This is done under several headings, corresponding to different ...

  20. Small Spacecraft Activities at JPL

    OpenAIRE

    Jones, Ross

    1991-01-01

    This paper presents a brief technical description of some of the small spacecraft concepts prepared by JPL for various sponsors. Some of JPL's work in microspacecraft is presented. The paper contains brief technical descriptions of the following four small spacecraft conceptual designs: 1) Lunar GAS, 2) Polar Mesoscale Explorer, 3) DARPA SHF and 4) Discovery. Since 1986, JPL has studied more than 10 small spacecraft including those to be presented here.

  1. Leadership in a small enterprise

    OpenAIRE

    Palmgren, Helena

    2010-01-01

    Leadership is often seen as the premier force behind the success and failure of enterprises and the well-being of the personnel. In spite of the vast amount of studies on leadership, leadership in small enterprises has seldom been in focus in leadership research. The increasing importance of small firms to the economic growth and competitiveness raises questions about the role and practice of leadership in small enterprises. This study aims to deepen our understanding of leadership as a socia...

  2. Organization of Small Business Accounting

    OpenAIRE

    Makarova, Oksana

    2012-01-01

    Along with continuing changes in the tax law, the requirements to the organization of small business accounting are changing, too. The article provides the analysis of normative legal documents governing small business accounting. The accounting requirements for small businesses of general and simplified tax accounts have been systematized. The necessity and essentiality of drawing primary documents has been grounded. The list and order of making tax registers and preparation of financial sta...

  3. Small Wind Site Assessment Guidelines

    Energy Technology Data Exchange (ETDEWEB)

    Olsen, Tim [Advanced Energy Systems LLC, Eugene, OR (United States); Preus, Robert [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2015-09-01

    Site assessment for small wind energy systems is one of the key factors in the successful installation, operation, and performance of a small wind turbine. A proper site assessment is a difficult process that includes wind resource assessment and the evaluation of site characteristics. These guidelines address many of the relevant parts of a site assessment with an emphasis on wind resource assessment, using methods other than on-site data collection and creating a small wind site assessment report.

  4. Deterministic Small-World Networks

    OpenAIRE

    Comellas, Francesc; Sampels, Michael

    2001-01-01

    Many real life networks, such as the World Wide Web, transportation systems, biological or social networks, achieve both a strong local clustering (nodes have many mutual neighbors) and a small diameter (maximum distance between any two nodes). These networks have been characterized as small-world networks and modeled by the addition of randomness to regular structures. We show that small-world networks can be constructed in a deterministic way. This exact approach permits a direct calculatio...

  5. Patient Safety Outcomes in Small Urban and Small Rural Hospitals

    Science.gov (United States)

    Vartak, Smruti; Ward, Marcia M.; Vaughn, Thomas E.

    2010-01-01

    Purpose: To assess patient safety outcomes in small urban and small rural hospitals and to examine the relationship of hospital and patient factors to patient safety outcomes. Methods: The Nationwide Inpatient Sample and American Hospital Association annual survey data were used for analyses. To increase comparability, the study sample was…

  6. Small white matter lesion detection in cerebral small vessel disease

    Science.gov (United States)

    Ghafoorian, Mohsen; Karssemeijer, Nico; van Uden, Inge; de Leeuw, Frank E.; Heskes, Tom; Marchiori, Elena; Platel, Bram

    2015-03-01

    Cerebral small vessel disease (SVD) is a common finding on magnetic resonance images of elderly people. White matter lesions (WML) are important markers for not only the small vessel disease, but also neuro-degenerative diseases including multiple sclerosis, Alzheimer's disease and vascular dementia. Volumetric measurements such as the "total lesion load", have been studied and related to these diseases. With respect to SVD we conjecture that small lesions are important, as they have been observed to grow over time and they form the majority of lesions in number. To study these small lesions they need to be annotated, which is a complex and time-consuming task. Existing (semi) automatic methods have been aimed at volumetric measurements and large lesions, and are not suitable for the detection of small lesions. In this research we established a supervised voxel classification CAD system, optimized and trained to exclusively detect small WMLs. To achieve this, several preprocessing steps were taken, which included a robust standardization of subject intensities to reduce inter-subject intensity variability as much as possible. A number of features that were found to be well identifying small lesions were calculated including multimodal intensities, tissue probabilities, several features for accurate location description, a number of second order derivative features as well as multi-scale annular filter for blobness detection. Only small lesions were used to learn the target concept via Adaboost using random forests as its basic classifiers. Finally the results were evaluated using Free-response receiver operating characteristic.

  7. Phosphatidylinositol-bisphosphate regulates intercellular coupling in cardiac myocytes

    DEFF Research Database (Denmark)

    Hofgaard, Johannes P; Banach, Kathrin; Mollerup, Sarah;

    2008-01-01

    that agonist-induced changes in PIP(2) can result in a reduction of the functional coupling of cardiomyocytes and, consequently, in changes in conduction velocity. Intercellular coupling was measured by Lucifer Yellow dye transfer in cultured neonatal rat cardiomyocytes. Conduction velocity was...

  8. Physical chemistry and membrane properties of two phosphatidylinositol bisphosphate isomers.

    Science.gov (United States)

    Slochower, David R; Wang, Yu-Hsiu; Radhakrishnan, Ravi; Janmey, Paul A

    2015-05-21

    The most highly charged phospholipids, polyphosphoinositides, are often involved in signaling pathways that originate at cell-cell and cell-matrix contacts, and different isomers of polyphosphoinositides have distinct biological functions that cannot be explained by separate highly specific protein ligand binding sites [Lemmon, Nat. Rev. Mol. Cell Biol., 2008, 9, 99-111]. PtdIns(3,5)P2 is a low abundance phosphoinositide localized to cytoplasmic-facing membrane surfaces, with relatively few known ligands, yet PtdIns(3,5)P2 plays a key role in controlling membrane trafficking events and cellular stress responses that cannot be duplicated by other phosphoinositides [Dove et al., Nature, 1997, 390, 187-192; Michell, FEBS J., 2013, 280, 6281-6294]. Here we show that PtdIns(3,5)P2 is structurally distinct from PtdIns(4,5)P2 and other more common phospholipids, with unique physical chemistry. Using multiscale molecular dynamics techniques on the quantum level, single molecule, and in bilayer settings, we found that the negative charge of PtdIns(3,5)P2 is spread over a larger area, compared to PtdIns(4,5)P2, leading to a decreased ability to bind divalent ions. Additionally, our results match well with experimental data characterizing the cluster forming potential of these isomers in the presence of Ca(2+) [Wang et al., J. Am. Chem. Soc., 2012, 134, 3387-3395; van den Bogaart et al., Nature, 2011, 479, 552-555]. Our results demonstrate that the different cellular roles of PtdIns(4,5)P2 and PtdIns(3,5)P2in vivo are not simply determined by their localization by enzymes that produce or degrade them, but also by their molecular size, ability to chelate ions, and the partial dehydration of those ions, which might affect the ability of PtdIns(3,5)P2 and PtdIns(4,5)P2 to form phosphoinositide-rich clusters in vitro and in vivo. PMID:25901568

  9. Targeted therapies in development for non-small cell lung cancer

    Directory of Open Access Journals (Sweden)

    Thanyanan Reungwetwattana

    2013-01-01

    Full Text Available The iterative discovery in various malignancies during the past decades that a number of aberrant tumorigenic processes and signal transduction pathways are mediated by "druggable" protein kinases has led to a revolutionary change in drug development. In non-small cell lung cancer (NSCLC, the ErbB family of receptors (e.g., EGFR [epidermal growth factor receptor], HER2 [human epidermal growth factor receptor 2], RAS (rat sarcoma gene, BRAF (v-raf murine sarcoma viral oncogene homolog B1, MAPK (mitogen-activated protein kinase c-MET (c-mesenchymal-epithelial transition, FGFR (fibroblast growth factor receptor, DDR2 (discoidin domain receptor 2, PIK3CA (phosphatidylinositol-4,5-bisphosphate3-kinase, catalytic subunit alpha, PTEN (phosphatase and tensin homolog, AKT (protein kinase B, ALK (anaplastic lym phoma kinase, RET (rearranged during transfection, ROS1 (reactive oxygen species 1 and EPH (erythropoietin-producing hepatoma are key targets of various agents currently in clinical development. These oncogenic targets exert their selective growth advantage through various intercommunicating pathways, such as through RAS/RAF/MEK, phosphoinositide 3-kinase/AKT/mammalian target of rapamycin and SRC-signal transduction and transcription signaling. The recent clinical studies, EGFR tyrosine kinase inhibitors and crizotinib were considered as strongly effective targeted therapies in metastatic NSCLC. Currently, five molecular targeted agents were approved for treatment of advanced NSCLC: Gefitinib, erlotinib and afatinib for positive EGFR mutation, crizotinib for positive echinoderm microtubule-associated protein-like 4 (EML4-ALK translocation and bevacizumab. Moreover, oncogenic mutant proteins are subject to regulation by protein trafficking pathways, specifically through the heat shock protein 90 system. Drug combinations affecting various nodes in these signaling and intracellular processes are predicted and demonstrated to be synergistic and

  10. Adenocarcinoma of the small bowel

    International Nuclear Information System (INIS)

    Adenocarcinoma of small bowel is generally a rather rare primary tumour of small bowel with a prevalence rate of 0.5 - 3.0 / 100.000 population, but the most frequent tumour of small intestine. It more often involves the duodenum and jejunum than the ileum. The aim of this paper is also to point out the value of small bowel follow through (SBFT) in the diagnosis of stenosing lesions. An 83 - year old male patient suffered from abdominal pain, malaise, vomiting, cachexia and diarrhoea for 3 months. The result of occult blood testing was negative. Haemoglobin level was normal. Proctoscopy, colonoscopy, upper gastrointestinal (GI) endoscopy, and ultrasonography (US) did not explain the patient's problems. Ileus of the small bowel was established with abdominal plain film. Small bowel follow through (SBFT) and computer tomography (CT) showed a stenosing tumour in the jejunum. Adenocarcinoma of the small bowel was established with histological examination after resection of the tumor. SBFT, with manual compression of all segments of the small bowel, can be a very accurate diagnostic investigation for evaluation of stenosing lesions in this part of the intestine. (author)

  11. What Is a Small Business?

    OpenAIRE

    Jerome S. Osteryoung; Newman, Derek

    1993-01-01

    This paper argues that a definition of small business is important for both research and governmental policy. A chronological history of definitions is discussed along with their limitations. A new definition of small business is advocated when a business has personal guarantees and has no public stock. This definition is measurable, meaningful, and congruent with the perceptions of the market system.

  12. Standard Deviation for Small Samples

    Science.gov (United States)

    Joarder, Anwar H.; Latif, Raja M.

    2006-01-01

    Neater representations for variance are given for small sample sizes, especially for 3 and 4. With these representations, variance can be calculated without a calculator if sample sizes are small and observations are integers, and an upper bound for the standard deviation is immediate. Accessible proofs of lower and upper bounds are presented for…

  13. Resourcing Change in Small Schools

    Science.gov (United States)

    Anderson, Michelle; White, Simone

    2011-01-01

    The theme of this article is the challenge that school leaders face in creating the conditions for learning in small schools. We draw on the concepts of "social capital" and "social entrepreneurship" to identify tensions and possibilities for school leaders in a case study of a small rural school as they seek to find resources for school-community…

  14. AcEST: DK960529 [AcEST

    Lifescience Database Archive (English)

    Full Text Available |TOR1A_MACFA Torsin-1A OS=Macaca fascicularis GN=TOR1A ... 33 1.7 sp|P85111|RCA_VITSX Ribulose bisphosp...|A9PFQ2_POPTR Putative uncharacterized protein OS=Popul... 228 2e-58 tr|Q06B51|Q06B51_ACERU Chloroplast ribulose-1,5-bisphosp...I 649 +K+GI Sbjct: 180 AKMGI 184 >tr|Q06B50|Q06B50_ACERU Chloroplast ribulose-1,5-bisphosphate carboxylase/o...9 Definition Adiantum capillus-veneris mRNA. clone: TST39A01NGRL0007_K16. 5' end sequence. Accession DK960529 Tissue type pr...|Q7X999|RCA2_LARTR Ribulose bisphosphate carboxylase/oxygenase activase 2, chloroplastic OS=Larrea tri

  15. AcEST: DK943967 [AcEST

    Lifescience Database Archive (English)

    Full Text Available 2. Query= DK943967|Adiantum capillus-veneris mRNA, clone: YMU02A01NGRL0004_I17, 5' (504 letters) Database: uniprot_spr...3|Y833_METJA Uncharacterized protein MJ0833 OS=Methanoca... 33 0.75 sp|O74252|STE12_EMENI Transcription factor steA OS=Emeri...Q06B48|Q06B48_ACERU Chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase activase large protein isofo...oxygenase ... 43 9e-09 sp|P58555|RCA_ANASP Ribulose bisphosphate carboxylase/oxygenase ... 37 3e-06 sp|Q5824...|Q674R7|ATG9B_HUMAN Autophagy-related protein 9B OS=Homo sapie... 30 8.3 >sp|P10896|RCA_ARATH Ribulose bisphosp

  16. AcEST: DK947455 [AcEST

    Lifescience Database Archive (English)

    Full Text Available |Q9BYV9|BACH2_HUMAN Transcription regulator protein BACH2 OS=H... 32 2.3 sp|P15257|HNF1A_RAT Hepatocyte nu...e-64 tr|Q19RP2|Q19RP2_9MAGN Ribulose-1,5-bisphosphate carboxylase/oxy... 246 7e-64 tr|A9NV87|A9NV87_PICSI Putative uncharacteri...ery= DK947455|Adiantum capillus-veneris mRNA, clone: YMU02A01NGRL0015_P10, 5' (58...|Q7X9A0|RCA1_LARTR Ribulose bisphosphate carboxylase/oxygenase activase 1, chloroplastic OS=Larrea tri...AST: a new generation of protein database search programs, Nucleic Acids Res. 25:3389-3402. Query= DK947455|Adiantum capillus-veneris

  17. Board Effectiveness in Small Firms

    OpenAIRE

    Söderqvist, Anette; WÀgar, Karolina

    2010-01-01

    Purpose: This study investigates boards of directors in small firms and explores the link between board effectiveness and the composition, roles and working styles of the boards. Design/methodology/approach: The study analyses data from a telephone survey of boards in 45 small firms. The survey included both the CEO and the chairperson of the board. Findings: The study identifies three groups of small firms: ‘paperboards’, ‘professional boards’, and ‘management ...

  18. Cosmic Strings with Small Tension

    CERN Document Server

    Halyo, Edi

    2009-01-01

    We describe cosmic F--term strings with exponentially small tension which are D3 branes wrapped on deformed $A_3$ singularities. We show that brane instanton effects which can be calculated after a geometric transition give rise to an exponentially small volume for the node on which the D3 branes wrap leading to a string with small tension. We generalize our description to the case of non--Abelian cosmic strings and argue that these strings are stable against monopole--anti monopole pair creation.

  19. Cytonuclear Evolution of Rubisco in Four Allopolyploid Lineages

    OpenAIRE

    Gong, Lei; Olson, Mischa; Wendel, Jonathan F.

    2014-01-01

    Allopolyploidization in plants entails the merger of two divergent nuclear genomes, typically with only one set (usually maternal) of parental plastidial and mitochondrial genomes and with an altered cytonuclear stoichiometry. Thus, we might expect cytonuclear coevolution to be an important dimension of allopolyploid evolution. Here, we investigate cytonuclear coordination for the key chloroplast protein rubisco (ribulose 1,5-bisphosphate carboxylase/oxygenase), which is composed of nuclear-e...

  20. SoyProLow: A protein database enriched in low abundant soybean proteins

    OpenAIRE

    Tavakolan, Mona; Alkharouf, Nadim W; Matthews, Benjamin F.; Natarajan, Savithiry S.

    2014-01-01

    Soybeans are an important legume crop that contain 2 major storage proteins, β-conglycinin and glycinin, which account about 70- 80% of total seed proteins. These abundant proteins hinder the isolation and characterization of several low abundant proteins in soybean seeds. Several protein extraction methodologies were developed in our laboratory to decrease these abundant storage proteins in seed extracts and to also decrease the amount of ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuB...

  1. Contrasting Diversity and Host Association of Ectomycorrhizal Basidiomycetes versus Root-Associated Ascomycetes in a Dipterocarp Rainforest

    OpenAIRE

    Sato, Hirotoshi; Tanabe, Akifumi S; Toju, Hirokazu

    2015-01-01

    Root-associated fungi, including ectomycorrhizal and root-endophytic fungi, are among the most diverse and important belowground plant symbionts in dipterocarp rainforests. Our study aimed to reveal the biodiversity, host association, and community structure of ectomycorrhizal Basidiomycota and root-associated Ascomycota (including root-endophytic Ascomycota) in a lowland dipterocarp rainforest in Southeast Asia. The host plant chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase large...

  2. Influence of temperature on measurements of the CO2 compensation point: differences between the Laisk and O2-exchange methods

    OpenAIRE

    Walker, Berkley J.; Cousins, Asaph B.

    2013-01-01

    The CO2 compensation point in the absence of day respiration (Γ*) is a key parameter for modelling leaf CO2 exchange. Γ* links the kinetics of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) with the stoichiometry of CO2 released per Rubisco oxygenation from photorespiration (α), two essential components of biochemical models of photosynthesis. There are two main gas-exchange methods for measuring Γ*: (i) the Laisk method, which requires estimates of mesophyll conductance to CO2 (g ...

  3. A Rubisco mutant that confers growth under a normally “inhibitory” oxygen concentration†

    OpenAIRE

    Satagopan, Sriram; Scott, Stephanie S.; Smith, Todd G.; Tabita, F. Robert

    2009-01-01

    Ribulose 1, 5-bisphosphate (RuBP) carboxylase/oxygenase (Rubisco) is a globally significant biocatalyst that facilitates the removal and sequestration of CO2 from the biosphere. Rubisco-catalyzed CO2 reduction thus provides virtually all the organic carbon utilized by living organisms. Despite catalyzing the rate-limiting step of photosynthetic and chemoautotrophic CO2 assimilation, Rubisco is markedly inefficient as the competition between O2 and CO2 for the same substrate limits the ability...

  4. PHYLOGENETIC ANALYSIS AMONG FOUR SECTIONS OF GENUS DENDROBIUM SW. (ORCHIDACEAE) IN PENINSULAR MALAYSIA USING RBCL SEQUENCE DATA

    OpenAIRE

    2013-01-01

    Phylogenetic analysis using chloroplast DNA, the ribulose-bisphosphate carboxylase gene (rbcL), was conducted to examine relationship among four sections of the genus Dendrobium (Orchidaceae): Aporum, Crumenata, Strongyle, and Bolbidium in Peninsular Malaysia. Classifications based on morphological characters have not been able to clearly divide these four sections, therefore deeper and detailed analyses are required to ascertain their status. In this study, the phylogenetic relationships amo...

  5. Isolation and kinetic characterisation of hydrophobically distinct populations of form I Rubisco

    OpenAIRE

    O’Donnelly, Kerry; Zhao, Guangyuan; Patel, Priya; Butt, M Salman; Mak, Lok Hang; Kretschmer, Simon; Woscholski, Rudiger; Barter, Laura M. C.

    2014-01-01

    Background Rubisco (Ribulose-1,5-bisphosphate carboxylase/oxygenase) is a Calvin Cycle enzyme involved in CO2 assimilation. It is thought to be a major cause of photosynthetic inefficiency, suffering from both a slow catalytic rate and lack of specificity due to a competing reaction with oxygen. Revealing and understanding the engineering rules that dictate Rubisco’s activity could have a significant impact on photosynthetic efficiency and crop yield. Results This paper describes the purifica...

  6. Isolation and Characterization of the Prochlorococcus Carboxysome Reveal the Presence of the Novel Shell Protein CsoS1D

    OpenAIRE

    Roberts, Evan W.; Cai, Fei; Cheryl A Kerfeld; Cannon, Gordon C.; Heinhorst, Sabine

    2012-01-01

    Cyanobacteria, including members of the genus Prochlorococcus, contain icosahedral protein microcompartments known as carboxysomes that encapsulate multiple copies of the CO2-fixing enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) in a thin protein shell that enhances the catalytic performance of the enzyme in part through the action of a shell-associated carbonic anhydrase. However, the exact mechanism by which compartmentation provides a catalytic advantage to the enzyme is ...

  7. Rubisco : easy purification and immunochemical determination

    OpenAIRE

    Groß, Ulrich

    2001-01-01

    Rubisco (Ribulose-1.5-bisphosphate carboxylase/oxygenase) from spinach was purified to homogeneity in one step by gel filtration. This enzyme is suitable for the generation of a specific antibody in rabbits. The enzyme concentration in spinach leaves amounted to 40 % of the total soluble protein. The specific antibody shows cross reaction with crude extracts from leaves of other higher plants. The enzymes subunits could be separated by denaturing preparative SDS gel electrophoresis.

  8. Photorespiration

    OpenAIRE

    Peterhansel, Christoph; Horst, Ina; Niessen, Markus; Blume, Christian; Kebeish, Rashad; Kürkcüoglu, Sophia; Kreuzaler, Fritz

    2010-01-01

    Photorespiration is initiated by the oxygenase activity of ribulose-1,5-bisphosphate-carboxylase/oxygenase (RUBISCO), the same enzyme that is also responsible for CO2 fixation in almost all photosynthetic organisms. Phosphoglycolate formed by oxygen fixation is recycled to the Calvin cycle intermediate phosphoglycerate in the photorespiratory pathway. This reaction cascade consumes energy and reducing equivalents and part of the afore fixed carbon is again released as CO2. Because of this, ph...

  9. Photorespiration: metabolic pathways and their role in stress protection

    OpenAIRE

    Wingler, A; Lea, P J; Quick, W.P.; Leegood, R C

    2000-01-01

    Photorespiration results from the oxygenase reaction catalysed by ribulose-1,5-bisphosphate carboxylase/oxygenase. In this reaction glycollate-2-phosphate is produced and subsequently metabolized in the photorespiratory pathway to form the Calvin cycle intermediate glycerate-3-phosphate. During this metabolic process, CO2 and NH3 are produced and ATP and reducing equivalents are consumed, thus making photorespiration a wasteful process. However, precisely because of this inefficiency, photore...

  10. Small Probe Reentry System Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Global Aerospace Corporation (GAC), and its research partner, Cal Poly San Luis Obispo (CPSLO), will develop an integrated Small Probe Reentry System (SPRS) for low...

  11. JPL Small Body Database Browser

    Data.gov (United States)

    National Aeronautics and Space Administration — The JPL Small-Body Database Browser provides data for all known asteroids and many comets. Newly discovered objects and their orbits are added on a daily basis....

  12. Small Sat Analysis Laboratory Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The NASA Low-Cost Small Spacecraft Program is focused on the technologies, subsystems, methodologies, and mission concepts for space missions which lower the...

  13. Small Sat Analysis Laboratory Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Develop Small Satellite Analysis Laboratory (SatLab): A simulation-of-simulations framework to integrate component and engineering simulations into a single larger...

  14. Quantum Small-world Networks

    CERN Document Server

    Wei, Zong-Wen; Han, Xiao-Pu

    2011-01-01

    Quantum networks are critical to quantum communication and distributed quantum computing. Here we propose a small-world model of large-scale quantum repeater networks, where "small-world" is a fundamental concept rooted in complex networks, which describe a broad range of real systems. The core of the model is to relate the hierarchical fashion of measurements to coarse-graining process, when quantum repeater protocols are implemented. We demonstrate that quantum repeater networks with fractal structure can be enlarged with certain length scale in geographic space, while preserving topology by performing renormalization. Actually, renormalization here serves as an organizing principle determining the distribution of long-range entangled links over quantum networks, which gives rise to fractal to small-world transition. Furthermore, by iterative implementation of renormalization on the former coarse-grained network, we eventually obtain an onion-like, hierarchical quantum small-world network, where the distanc...

  15. Development project of small accelerator

    CERN Document Server

    Yamada, S

    2002-01-01

    The object of this project is demonstration of a small proton and heavy ion synchrotron and a small hard X-ray photon radiation source by using new technology and application of them to therapy, diagnosis, material science and life science. In this paper, a part of small proton and heavy ion synchrotron is discussed. Nine organizations joined in this project. There are four development themes such as optimization of laser-ion 100 TW class source target, beam storage and cooling device, small synchrotron ring and FFAG accelerator. Outline and contents of development of them are explained. This project is planning to generate a few MeV/u carbon ions in fully ionized states by impact of laser with about 100 TW output. 3 T maximum bending magnetic field using normal conduction AC magnet will be actualized for synchrotron with 200 MeV proton beam. (S.Y.)

  16. Small for gestational age (SGA)

    Science.gov (United States)

    ... is called intrauterine growth restriction. The most common definition of small for gestational age (SGA) is a ... M. is also a founding member of Hi-Ethics and subscribes to the principles of the Health ...

  17. Business Opportunities for Small Reactors

    International Nuclear Information System (INIS)

    This report assesses the market potential and identifies a number of potential paths for developing the small nuclear reactor business. There are several potential opportunities identified and evaluated. Selecting a specific approach for the business development requires additional information related to a specific market and sources of capital to support the investment. If and how a market for small nuclear plants may develop is difficult to predict because of the complexity of the economic and institutional factors that will influence such development. Key factors are; economics, safety, proliferation resistance and investment risk. The economic and political interest of any of the identified markets is also dependent on successful demonstration of the safety and reliability of small nuclear reactor. Obtaining a US-NRC Standard Design approval would be an important development step toward establishing a market for small reactors. (authors)

  18. Small satellites and their regulation

    CERN Document Server

    Jakhu, Ram S

    2014-01-01

    Since the launch of UoSat-1 of the University of Surrey (United Kingdom) in 1981, small satellites proved regularly to be useful, beneficial, and cost-effective tools. Typical tasks cover education and workforce development, technology demonstration, verification and validation, scientific and engineering research as well as commercial applications. Today the launch masses range over almost three orders of magnitude starting at less than a kilogram up to a few hundred kilograms, with budgets of less than US$ 100.00 and up to millions within very short timeframes of sometimes less than two years. Therefore each category of small satellites provides specific challenges in design, development and operations. Small satellites offer great potentials to gain responsive, low-cost access to space within a short timeframe for institutions, companies, regions and countries beyond the traditional big players in the space arena. For these reasons (particularly the low cost of construction, launch and operation), small (m...

  19. Very small ionization pressure gauge

    International Nuclear Information System (INIS)

    A very small ionization pressure gauge has been developed to operate in the pressure range 10-6 Torr to 100μ. A metal construction and external cooling fins result in a very rugged device with exceptionally small outgassing properties. The gauge also incorporates a replaceable filament-grid assembly on a single plug which allows the simple replacement of both the filament and the grid when needed

  20. Ecology in Small Aquatic Ecosystems

    DEFF Research Database (Denmark)

    Andersen, Mikkel René

    % supersaturated in oxygen while the bottom waters becomes anoxic. Dense charophyte stands influenced the hydrodynamics and created favorable conditions for the apical parts in the surface waters, while the basal parts withstood anoxia for up to 12 hours in the bottom waters. Nocturnal convective mixing oxygenated......) both in the small lakes and in the small ephemeral streams on the Great Alvar. Downstream respiration was decoupled from GPP as respiration rates were much higher due to agricultural impact....

  1. Small Bowel Review - Part I

    OpenAIRE

    Thomson, ABR; Wild, G.

    1997-01-01

    Significant advances have been made in the study of the small bowel. Part I of this two-part review of the small bowel examines carbohydrates, including brush border membrane hydrolysis and sugar transport; amino acids, dipeptides, proteins and food allergy, with a focus on glutamine, peptides and macromolecules, and nucleosides, nucleotides and polyamines; salt and water absorption, and diarrhea, including antidiarrheal therapy and oral rehydration treatment; lipids (digestion and absorption...

  2. Small windpower systems are beautiful

    Energy Technology Data Exchange (ETDEWEB)

    Conway, A.

    1988-11-01

    Today's small wind turbine generators perform well and reliably, thanks to brushless alternators, composite blades and protection against high winds. Wind turbines of such dimensions would often be employed for battery chargers. Bigger, geared machines, with rotors of up to 17 m diameter, driving synchronous or induction generators, would be coupled with diesel sets for supplying small communities. Battery chargers, island and hybrid systems and possible sites are discussed.

  3. Unitarity at small Bjorken x

    OpenAIRE

    Weigert, Heribert

    2000-01-01

    This paper presents a solution to the nonlinear small x ``projectile side'' evolution equations as derived by Balitskii in 1996. The solution is based on functional Fokker-Planck methods. The fixed point at small x is explicitly calculated and all correlation functions in this limit are determined. They show clear saturation and unitarization properties. Scaling laws that hold during the saturation phase and throughout the whole course of the evolution are established. The corresponding Lange...

  4. Effect of fructose-1,6-bisphosphate on the myocardial ultrastructure and oxidative stress in Diabetic cardiomyopathy rats%1,6-二磷酸果糖对糖尿病大鼠心肌超微结构及氧化应激的影响

    Institute of Scientific and Technical Information of China (English)

    孙冬梅; 薛瑞凤; 郭淑香; 刘洋; 孙萌

    2010-01-01

    Objective To study the effect of Fructose-1,6-Bisphosphate ( FBP) on the myocardial ultrastructure and oxidative stress in rats with STZ-induced diabetic cardiomyopathy(DCM). Methods Diabetes mellitus model in healthy male Sprague-Dawley(SD) rats was induced by single injection of streptozotocin (STZ,Sigma)into intraperitoneal with a dose of 65mg/kg. Intervening subjects( FBP group) were given 500 mg/( kg · d) body weight of FBP intraperitoneal administration, while diabetes mellitus subjects ( DM group) and normal control subjects ( NC group) received 500 mg/( kg · d) of saline After one month, all rats were killed and the myocardial ultrastructures were studied by transmission electron microscopy (TEM). The contents of malondialdehyde(MDA) , activity of superoxide dismutase( SOD), nitric oxide synthase (NOS) and inductive nitric oxide synthase (iNOS) were measured in myocardial homogenate. Results Compared with NC group, more ruptured myofibril was found in DM group, as well as increased mitochondria, vacuolar degeneration and increased electron concentration of capillary basement membrane. At the same time, the concentration of MDA and activation of NOS and iNOS increased (P <0.05) , while the activation of NOS and iNOS in FBP group decreased (P < 0. 05). Conclusion FBP can protect diabetic myocardium and decrease oxidative stress.%目的 观察1,6-二磷酸果糖(FBP)对糖尿病大鼠心肌超微结构、氧化应激的影响.方法 将25只8周龄SPF级健康雄性SD大鼠按抽签法选择19只建立SD大鼠糖尿病模型,余下6只作为正常组.将造模成功的大鼠完全随机分成糖尿病组和FBP干预组(干预组,FBP 500 mg/kg·d腹腔注射),糖尿病组及正常组给予等量生理盐水腹腔注射.1个月后处死大鼠,取心肌组织进行透射电镜(TEM)观察,并测定心肌组织内氧化应激指标后进行比较.结果 与正常组相比,糖尿病组大鼠心肌肌原纤维断裂,线粒体增多、空泡变性,毛细血管基

  5. World Small Hydropower Development Report

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Heng; Esser, Lara (ICSGP (China)); Masera, Diego (UNIDO, Vienna (Austria))

    2013-07-01

    Currently, small hydropower plants with a capacity of 10 MW, exist in 148 countries or territories worldwide. Four other countries have been identified with resource potential. This report aims to identify the development status and resource potential of small hydro in various countries, territories and regions throughout the world. Working with experts at the ground level to compile and share existing information, experiences and challenges, one comprehensive report was created. Decision-makers, stakeholders and potential investors clearly need this comprehensive information to more effectively promote small hydropower as a renewable and rural energy source for sustainable development and to overcome the existing development barriers. The findings of this report show that small hydropower potential globally is approximated at almost 173 GW. The figure is arrived by totaling data from a wide range of sources with potential compromise of data integrity to varying degrees. For example, research data on economically feasible potential were more readily available in developed countries than those in the least developed or developing countries. More than half of the world's known hydropower potential is located in Asia, around one third can be found in Europe and the Americas. It is possible in the future that more small hydropower potential might be identified both on the African and American continents. The installed small hydropower capacity (up to 10 MW) is estimated to be 75 GW in 2011/2012. The report provides detailed data for each country/region, including recommendations on the national, regional and international level.

  6. Small

    Energy Technology Data Exchange (ETDEWEB)

    Montoya, Joseph

    2013-07-18

    Representing the Center on Nanostructuring for Efficient Energy Conversion (CNEEC), this document is one of the entries in the Ten Hundred and One Word Challenge. As part of the challenge, the 46 Energy Frontier Research Centers were invited to represent their science in images, cartoons, photos, words and original paintings, but any descriptions or words could only use the 1000 most commonly used words in the English language, with the addition of one word important to each of the EFRCs and the mission of DOE energy. The mission of CNEEC is to understand how nanostructuring can enhance efficiency for energy conversion and solve fundamental cross-cutting problems in advanced energy conversion and storage systems.

  7. 78 FR 48537 - Small Business Innovation Research and Small Business Technology Transfer Programs...

    Science.gov (United States)

    2013-08-08

    ... ADMINISTRATION Small Business Innovation Research and Small Business Technology Transfer Programs... Administration (SBA) is publishing the Small Business Innovation Research (SBIR) and Small Business Technology... Director, Office of Innovation, Small Business Administration, 409 Third Street SW., Washington, DC...

  8. 78 FR 59410 - Small Business Innovation Research and Small Business Technology Transfer Programs...

    Science.gov (United States)

    2013-09-26

    ... ADMINISTRATION Small Business Innovation Research and Small Business Technology Transfer Programs... period for the Small Business Innovation Research (SBIR) and Small Business Technology Transfer (STTR... Edsel Brown, Jr., Assistant Director, Office of Innovation, Small Business Administration, 409...

  9. Runoff processes and small watersheds

    International Nuclear Information System (INIS)

    Full text: Small watersheds are a fundamental landscape unit for quantifying inputs and outputs of water, sediment and nutrients. Small watersheds have been used historically for defining runoff processes and flood response to storm precipitation. Early conceptualizations of runoff production during the International Hydrological Decade in the 1960s focused on the importance and movement of event water as overland flow to the stream channel. Use of mass balance mixing models using stable isotope tracers in the 1970s and 1980s directly challenged early ideas of where water goes when it rains, residence time of catchment waters and flow paths of subsurface runoff towards the stream. These data showed that the majority of water in the stream during a precipitation event was water that existed in the watershed prior to the event. While credible physical mechanisms of old water mobilization have only been defined in the past decade, stable isotope tracer approaches are now mature enough to offer new potential for informing new model structures of how small watersheds work. Isotope tracer data in small watersheds and mass balance separations also represent new ways of validating and calibrating watershed models. This presentation will chronicle the use of isotope tracers in small watersheds and provide examples of how these data can be used in models of runoff processes and for providing valuable input for water resource management at larger basin scales. (author)

  10. Small satellites for tropical applications

    Science.gov (United States)

    Montpetit, Marie-Jose; Bonn, Ferdinand

    1993-11-01

    A number of mission studies were performed to assess the suitability of small satellite systems for tropical data acquisition. These studies took into account the specifics of the tropical user communities and were focused on remote sensing and resource management issues. The requirements and potential solutions for four application areas are discussed. For monitoring of forest and agricultural vegetation, a small synthetic aperture radar is considered with P, C, or X band imaging, possibly supplemented by a high resolution multispectral imager. The radar would have the capability to monitor below cloud cover which is often found in tropical regions. Optical, microwave, or spectrographic imaging would also be useful in small satellites for disaster monitoring (notably of floods), land management, and air pollution monitoring. A small satellite with data storage and forwarding capability is also envisioned to collect data from dependable, low-power, and low-cost ground sensors via a simple ultrahigh frequency uplink and download the data on a very high frequency downlink. All the small satellites would be launched in low inclination orbits to ensure a number of consecutive passes over the targeted tropical area.

  11. Compound 13, an α1-selective small molecule activator of AMPK, inhibits Helicobacter pylori-induced oxidative stresses and gastric epithelial cell apoptosis

    International Nuclear Information System (INIS)

    Half of the world's population experiences Helicobacter pylori (H. pylori) infection, which is a main cause of gastritis, duodenal and gastric ulcer, and gastric cancers. In the current study, we investigated the potential role of compound 13 (C13), a novel α1-selective small molecule activator of AMP-activated protein kinase (AMPK), against H. pylori-induced cytotoxicity in cultured gastric epithelial cells (GECs). We found that C13 induced significant AMPK activation, evidenced by phosphorylation of AMPKα1 and ACC (acetyl-CoA carboxylase), in both primary and transformed GECs. Treatment of C13 inhibited H. pylori-induced GEC apoptosis. AMPK activation was required for C13-mediated GEC protection. Inhibition of AMPK kinase activity by the AMPK inhibitor Compound C, or silencing AMPKα1 expression by targeted-shRNAs, alleviated C13-induced GEC protective activities against H. pylori. Significantly, C13 inhibited H. pylori-induced reactive oxygen species (ROS) production in GECs. C13 induced AMPK-dependent expression of anti-oxidant gene heme oxygenase (HO-1) in GECs. Zinc protoporphyrin (ZnPP) and tin protoporphyrin (SnPP), two HO-1 inhibitors, not only suppressed C13-mediated ROS scavenging activity, but also alleviated its activity in GECs against H. pylori. Together, these results indicate that C13 inhibits H. pylori-induced ROS production and GEC apoptosis through activating AMPK–HO–1 signaling. - Highlights: • We synthesized compound 13 (C13), a α1-selective small molecule AMPK activator. • C13-induced AMPK activation requires α1 subunit in gastric epithelial cells (GECs). • C13 enhances Helicobacter pylori-induced pro-survival AMPK activation to inhibit GEC apoptosis. • C13 inhibits H. pylori-induced reactive oxygen species (ROS) production in GECs. • AMPK-heme oxygenase (HO-1) activation is required for C13-mediated anti-oxidant activity

  12. Compound 13, an α1-selective small molecule activator of AMPK, inhibits Helicobacter pylori-induced oxidative stresses and gastric epithelial cell apoptosis

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Hangyong; Zhu, Huanghuang; Lin, Zhou; Lin, Gang; Lv, Guoqiang, E-mail: lvguoqiangwuxivip@163.com

    2015-08-07

    Half of the world's population experiences Helicobacter pylori (H. pylori) infection, which is a main cause of gastritis, duodenal and gastric ulcer, and gastric cancers. In the current study, we investigated the potential role of compound 13 (C13), a novel α1-selective small molecule activator of AMP-activated protein kinase (AMPK), against H. pylori-induced cytotoxicity in cultured gastric epithelial cells (GECs). We found that C13 induced significant AMPK activation, evidenced by phosphorylation of AMPKα1 and ACC (acetyl-CoA carboxylase), in both primary and transformed GECs. Treatment of C13 inhibited H. pylori-induced GEC apoptosis. AMPK activation was required for C13-mediated GEC protection. Inhibition of AMPK kinase activity by the AMPK inhibitor Compound C, or silencing AMPKα1 expression by targeted-shRNAs, alleviated C13-induced GEC protective activities against H. pylori. Significantly, C13 inhibited H. pylori-induced reactive oxygen species (ROS) production in GECs. C13 induced AMPK-dependent expression of anti-oxidant gene heme oxygenase (HO-1) in GECs. Zinc protoporphyrin (ZnPP) and tin protoporphyrin (SnPP), two HO-1 inhibitors, not only suppressed C13-mediated ROS scavenging activity, but also alleviated its activity in GECs against H. pylori. Together, these results indicate that C13 inhibits H. pylori-induced ROS production and GEC apoptosis through activating AMPK–HO–1 signaling. - Highlights: • We synthesized compound 13 (C13), a α1-selective small molecule AMPK activator. • C13-induced AMPK activation requires α1 subunit in gastric epithelial cells (GECs). • C13 enhances Helicobacter pylori-induced pro-survival AMPK activation to inhibit GEC apoptosis. • C13 inhibits H. pylori-induced reactive oxygen species (ROS) production in GECs. • AMPK-heme oxygenase (HO-1) activation is required for C13-mediated anti-oxidant activity.

  13. Industry Research and Recommendations for Small Buildings and Small Portfolios

    Energy Technology Data Exchange (ETDEWEB)

    Langner, Rois [National Renewable Energy Lab. (NREL), Golden, CO (United States); Hendron, Bob [National Renewable Energy Lab. (NREL), Golden, CO (United States); Pless, Shanti [National Renewable Energy Lab. (NREL), Golden, CO (United States); Huppert, Mark [National Trust for Historic Preservation, Washington, DC (United States); Cochrane, Ric [National Trust for Historic Preservation, Washington, DC (United States)

    2013-12-01

    Small buildings have been left behind in the energy efficiency marketplace because financial and technical resources have flowed to larger commercial buildings. DOE's Building Technologies Office works with the commercial building industry to accelerate the uptake of energy efficiency technologies and techniques in existing and new commercial buildings (DOE 2013). BTO recognizes the SBSP sector'spotential for significant energy savings and the need for investments in resources that are tailored to this sector's unique needs. The industry research and recommendations described in this report identify potential approaches and strategic priorities that BTO could explore over the next 3-5 years that will support the implementation of high-potential energy efficiency opportunities for thisimportant sector. DOE is uniquely positioned to provide national leadership, objective information, and innovative tools, technologies, and services to support cost-effective energy savings in the fragmented and complex SBSP sector. Properly deployed, the DOE effort could enhance and complement current energy efficiency approaches. Small portfolios are loosely and qualitatively defined asportfolios of buildings that include only a small number of small buildings. This distinction is important because the report targets portfolio owners and managers who generally do not have staff and other resources to track energy use and pursue energy efficiency solutions.

  14. Marketing the Uniqueness of Small Towns. Small Town Strategy.

    Science.gov (United States)

    Hogg, David H.; Dunn, Douglas

    A small town can strengthen its local economy as a result of business people and concerned citizens collectively identifying that community's uniqueness and then capitalizing on it via advertising, personal selling, sales promotion, or publicity. This publication relates the science of marketing to communities. Seven simple techniques are provided…

  15. Studies in small field inflation

    International Nuclear Information System (INIS)

    We explore some issues in slow roll inflation in situations where field excursions are small compared to Mp. We argue that for small field inflation, minimizing fine tuning requires low energy supersymmetry and a tightly constrained structure. Hybrid inflation is almost an inevitable outcome. The resulting theory can be described in terms of a supersymmetric low energy effective action and inflation completely characterized in terms of a small number of parameters. Demanding slow roll inflation significantly constrains these parameters. In this context, the generic level of fine tuning can be described as a function of the number of light fields, there is an upper bound on the scale of inflation, and an (almost) universal prediction for the spectral index. Models of this type need not suffer from a cosmological moduli problem

  16. Small Business Social Responsibility Communication

    DEFF Research Database (Denmark)

    Morsing, Mette; Spence, Laura J.

    2015-01-01

    Purpose: Corporate social responsibility communication by small and medium sized enterprises is theorized to form the concept of Small Business Social Responsibility (SBSR) Communication. Design/methodology/approach: This is a conceptual paper that draws on Foucault’s theory of governmentality to....../implications: The conceptualization of SBSR communication would be enhanced by in-depth, nuanced empirical research. Practical implications: This paper has policy implications for those seeking to understand CSR among SMEs using formal communication mechanisms as a metric and control. Our paper suggests this to be...... intrinsic motivations) are confronting owner-managers’ ethos. We explain theoretically how small business managers respond to the challenge when they are required to formalize and display for external surveillance that which would otherwise be informal and part of the non-public or private sphere....

  17. Studies in Small Field Inflation

    CERN Document Server

    Dine, Michael

    2011-01-01

    We explore some issues in slow roll inflation in situations where field excursions are small compared to $M_p$. We argue that for small field inflation, minimizing fine tuning requires low energy supersymmetry and a tightly constrained structure. Hybrid inflation is {\\it almost} an inevitable outcome. The resulting theory can be described in terms of a supersymmetric low energy effective action and inflation completely characterized in terms of a small number of parameters. Demanding slow roll inflation significantly constrains these parameters. In this context, the generic level of fine tuning can be described as a function of the number of light fields, there is an upper bound on the scale of inflation, and an (almost) universal prediction for the spectral index. Models of this type need not suffer from a cosmological moduli problem.

  18. Trust Propagation in Small Worlds

    DEFF Research Database (Denmark)

    Gray, Elizabeth; Seigneur, Jean-Marc; Chen, Yong; Jensen, Christian D.

    do not scale well. We aim to develop trust-based security mechanisms using small world concepts to optimise formation and propagation of trust amongst entities in these vast networks. In this regard, we surmise that in a very large mobile ad hoc network, trust, risk, and recommendations can be...... propagated through relatively short paths connecting entities. Our work describes the design of trust-formation and risk-assessment systems, as well as that of an entity recognition scheme, within the context of the small world network topology....

  19. Small Wind Turbine Technology Assessment

    International Nuclear Information System (INIS)

    The result of the study carried out under the scope of the ATYCA project Test Plant of Wind Systems for Isolated Applications, about the state of art of the small wind turbine technology (wind turbines with swept area smaller than 40 m2) is presented. The study analyzes the collected information on 60 models of wind turbines from 23 manufacturers in the worldwide market. Data from Chinese manufacturers, that have a large participation in the total number of small wind turbines in operation, are not included, due to the unavailability of the technical information. (Author) 15 refs

  20. Orbit Mechanics about Small Asteroids

    Science.gov (United States)

    Scheeres, D. J.

    2007-01-01

    Space missions to small solar system bodies must deal with multiple perturbations acting on the spacecraft. These include strong perturbations from the gravity field and solar tide, but for small bodies the most important perturbations may arise from solar radiation pressure (SRP) acting on the spacecraft. Previous research has generally investigated the effect of the gravity field, solar tide, and SRP acting on a spacecraft trajectory about an asteroid in isolation and has not considered their joint effect. In this paper a more general theoretical discussion of the joint effects of these forces is given.

  1. Particles in small volume injections.

    Science.gov (United States)

    Taylor, S A; Spence, J

    1983-12-01

    The level of particulate contamination in small volume injections has been examined using the light blockage (HIAC) and electrical sensing zone (Coulter counter) techniques, the HIAC system being found to be the more suitable. Particle counts on the same batch of injection showed a large and variable difference between the HIAC and the Coulter counter results, especially below 5 micron. None of the injections examined complied with the British Pharmacopoeia limits for particulates in large volume parenterals, suggesting the unsuitability of the limits for small volume parenterals. PMID:6141237

  2. The growth of soybean under free air [CO(2)] enrichment (FACE) stimulates photosynthesis while decreasing in vivo Rubisco capacity.

    Science.gov (United States)

    Bernacchi, Carl J; Morgan, Patrick B; Ort, Donald R; Long, Stephen P

    2005-01-01

    Down-regulation of light-saturated photosynthesis (A(sat)) at elevated atmospheric CO(2) concentration, [CO(2)], has been demonstrated for many C(3) species and is often associated with inability to utilize additional photosynthate and/or nitrogen limitation. In soybean, a nitrogen-fixing species, both limitations are less likely than in crops lacking an N-fixing symbiont. Prior studies have used controlled environment or field enclosures where the artificial environment can modify responses to [CO(2)]. A soybean free air [CO(2)] enrichment (FACE) facility has provided the first opportunity to analyze the effects of elevated [CO(2)] on photosynthesis under fully open-air conditions. Potential ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylation (V(c,max)) and electron transport through photosystem II (J(max)) were determined from the responses of A(sat) to intercellular [CO(2)] (C(i)) throughout two growing seasons. Mesophyll conductance to CO(2) (g(m)) was determined from the responses of A(sat) and whole chain electron transport (J) to light. Elevated [CO(2)] increased A(sat) by 15-20% even though there was a small, statistically significant, decrease in V(c,max). This differs from previous studies in that V(c,max)/J(max) decreased, inferring a shift in resource investment away from Rubisco. This raised the C(i) at which the transition from Rubisco-limited to ribulose-1,5-bisphosphate regeneration-limited photosynthesis occurred. The decrease in V(c,max) was not the result of a change in g(m), which was unchanged by elevated [CO(2)]. This first analysis of limitations to soybean photosynthesis under fully open-air conditions reveals important differences to prior studies that have used enclosures to elevate [CO(2)], most significantly a smaller response of A(sat) and an apparent shift in resources away from Rubisco relative to capacity for electron transport. PMID:15252734

  3. Small molecule inhibition of 6-phosphofructo-2-kinase suppresses t cell activation

    Directory of Open Access Journals (Sweden)

    Telang Sucheta

    2012-05-01

    Full Text Available Abstract Background T cell activation is associated with a rapid increase in intracellular fructose-2,6-bisphosphate (F2,6BP, an allosteric activator of the glycolytic enzyme, 6-phosphofructo-1-kinase. The steady state concentration of F2,6BP in T cells is dependent on the expression of the bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases (PFKFB1-4 and the fructose-2,6-bisphosphatase, TIGAR. Of the PFKFB family of enzymes, PFKFB3 has the highest kinase:bisphosphatase ratio and has been demonstrated to be required for T cell proliferation. A small molecule antagonist of PFKFB3, 3-(3-pyridinyl-1-(4-pyridinyl-2-propen-1-one (3PO, recently has been shown to reduce F2,6BP synthesis, glucose uptake and proliferation in transformed cells. We hypothesized that the induction of PFKFB3 expression may be required for the stimulation of glycolysis in T cells and that exposure to the PFKFB3 antagonist, 3PO, would suppress T cell activation. Methods We examined PFKFB1-4 and TIGAR expression and F2,6BP concentration in purified CD3+ T cells stimulated with microbead-conjugated agonist antibodies specific for CD3 and the co-stimulatory receptor, CD28. We then determined the effect of 3PO on anti-CD3/anti-CD28-induced T cell activation, F2,6BP synthesis, 2-[1-14C]-deoxy-d-glucose uptake, lactate secretion, TNF-α secretion and proliferation. Finally, we examined the effect of 3PO administration on the development of delayed type hypersensitivity to methylated BSA and on imiquimod-induced psoriasis in mice. Results We found that purified human CD3+ T cells express PFKFB2, PFKFB3, PFKFB4 and TIGAR, and that anti-CD3/anti-CD28 conjugated microbeads stimulated a >20-fold increase in F2,6BP with a coincident increase in protein expression of the PFKFB3 family member and a decrease in TIGAR protein expression. We then found that exposure to the PFKFB3 small molecule antagonist, 3PO (1–10 μM, markedly attenuated the stimulation of F2,6BP

  4. Small Business Administration Semiannual Regulatory Agenda

    Science.gov (United States)

    2010-04-26

    ... Part XVI Small Business Administration Semiannual Regulatory Agenda ] SMALL BUSINESS ADMINISTRATION (SBA) SMALL BUSINESS ADMINISTRATION 13 CFR Ch. I Semiannual Regulatory Agenda AGENCY: U.S. Small... General Counsel for Legislation and Appropriations, U.S. Small Business Administration, 409 Third...

  5. Small Parts Assembler Work Sample.

    Science.gov (United States)

    Shawsheen Valley Regional Vocational-Technical High School, Billerica, MA.

    This manual contains a work sample intended to assess a handicapped student's interest in and potential to enter a training program in small parts assembly or in a similar job. Section 1 describes the assessment, correlates the work performed and worker traits required for completing the work sample, and lists related occupations and DOT codes.…

  6. Small Business. Lesson Plan Three.

    Science.gov (United States)

    Dicke, Thomas S.

    1996-01-01

    Presents a lesson plan that requires students to use their research and analytical skills in studying the social and economic importance of small business. Activities include interviewing local business owners and producing a written summary, developing a survey showing and explaining area land use, and researching opposition to Wal-Mart stores.…

  7. Joint research using small tokamaks

    Czech Academy of Sciences Publication Activity Database

    Gryaznevich, M.P.; Del Bosco, E.; Malaquias, A.; Mank, G.; Van Oost, G.; He, Yexi; Hegazy, H.; Hirose, A.; Hron, Martin; Kuteev, B.; Ludwig, G.O.; Nascimento, I.C.; Silva, C.; Vorobyev, G.M.

    2005-01-01

    Roč. 45, č. 10 (2005), S245-S254. ISSN 0029-5515. [Fusion Energy Conference contributions. Vilamoura, 1.11.2004-6.11.2004] Institutional research plan: CEZ:AV0Z20430508 Keywords : small tokamaks * thermonuclear fusion Subject RIV: BL - Plasma and Gas Discharge Physics Impact factor: 3.418, year: 2005

  8. Small Business Innovations (Crystal Components)

    Science.gov (United States)

    1991-01-01

    Scientific Materials Corporation, Bozeman, MT developed the SciMax line of improved Nd:Yag crystals under an Small Business Innovation Research (SBIR) contract with Langley Research Center. They reduced the amount of water trapped in the crystals during growth to improve the optical quality and efficiency. Applications of the crystals include fiber optics, telecommunications, welding, drilling, eye surgery and medical instrumentation.

  9. VEGA, a small launch vehicle

    Science.gov (United States)

    Duret, François; Fabrizi, Antonio

    1999-09-01

    Several studies have been performed in Europe aiming to promote the full development of a small launch vehicle to put into orbit one ton class spacecrafts. But during the last ten years, the european workforce was mainly oriented towards the qualification of the heavy class ARIANE 5 launch vehicle.Then, due also to lack of visibility on this reduced segment of market, when comparing with the geosatcom market, no proposal was sufficiently attractive to get from the potentially interrested authorities a clear go-ahead, i.e. a financial committment. The situation is now rapidly evolving. Several european states, among them ITALY and FRANCE, are now convinced of the necessity of the availability of such a transportation system, an important argument to promote small missions, using small satellites. Application market will be mainly scientific experiments and earth observation; some telecommunications applications may be also envisaged such as placement of little LEO constellation satellites, or replacement after failure of big LEO constellation satellites. FIAT AVIO and AEROSPATIALE have proposed to their national agencies the development of such a small launch vehicle, named VEGA. The paper presents the story of the industrial proposal, and the present status of the project: Mission spectrum, technical definition, launch service and performance, target development plan and target recurring costs, as well as the industrial organisation for development, procurement, marketing and operations.

  10. Small animal radiotherapy research platforms

    International Nuclear Information System (INIS)

    Advances in conformal radiation therapy and advancements in pre-clinical radiotherapy research have recently stimulated the development of precise micro-irradiators for small animals such as mice and rats. These devices are often kilovolt x-ray radiation sources combined with high-resolution CT imaging equipment for image guidance, as the latter allows precise and accurate beam positioning. This is similar to modern human radiotherapy practice. These devices are considered a major step forward compared to the current standard of animal experimentation in cancer radiobiology research. The availability of this novel equipment enables a wide variety of pre-clinical experiments on the synergy of radiation with other therapies, complex radiation schemes, sub-target boost studies, hypofractionated radiotherapy, contrast-enhanced radiotherapy and studies of relative biological effectiveness, to name just a few examples. In this review we discuss the required irradiation and imaging capabilities of small animal radiation research platforms. We describe the need for improved small animal radiotherapy research and highlight pioneering efforts, some of which led recently to commercially available prototypes. From this, it will be clear that much further development is still needed, on both the irradiation side and imaging side. We discuss at length the need for improved treatment planning tools for small animal platforms, and the current lack of a standard therein. Finally, we mention some recent experimental work using the early animal radiation research platforms, and the potential they offer for advancing radiobiology research. (topical review)

  11. Small animal radiotherapy research platforms

    Science.gov (United States)

    Verhaegen, Frank; Granton, Patrick; Tryggestad, Erik

    2011-06-01

    Advances in conformal radiation therapy and advancements in pre-clinical radiotherapy research have recently stimulated the development of precise micro-irradiators for small animals such as mice and rats. These devices are often kilovolt x-ray radiation sources combined with high-resolution CT imaging equipment for image guidance, as the latter allows precise and accurate beam positioning. This is similar to modern human radiotherapy practice. These devices are considered a major step forward compared to the current standard of animal experimentation in cancer radiobiology research. The availability of this novel equipment enables a wide variety of pre-clinical experiments on the synergy of radiation with other therapies, complex radiation schemes, sub-target boost studies, hypofractionated radiotherapy, contrast-enhanced radiotherapy and studies of relative biological effectiveness, to name just a few examples. In this review we discuss the required irradiation and imaging capabilities of small animal radiation research platforms. We describe the need for improved small animal radiotherapy research and highlight pioneering efforts, some of which led recently to commercially available prototypes. From this, it will be clear that much further development is still needed, on both the irradiation side and imaging side. We discuss at length the need for improved treatment planning tools for small animal platforms, and the current lack of a standard therein. Finally, we mention some recent experimental work using the early animal radiation research platforms, and the potential they offer for advancing radiobiology research.

  12. Sonography of the small intestine

    Institute of Scientific and Technical Information of China (English)

    Kim Nylund; Svein (φ)degaard; Trygve Hausken; Geir Folvik; Gülen Arslan Lied; Ivan Viola; Helwig Hauser; Odd-Helge Gilja

    2009-01-01

    In the last two decades, there has been substantial development in the diagnostic possibilities for examining the small intestine. Compared with computerized tomography, magnetic resonance imaging, capsule endoscopy and double-balloon endoscopy, ultrasonography has the advantage of being cheap, portable, flexible and user- and patient-friendly, while at the same time providing the clinician with image data of high temporal and spatial resolution. The method has limitations with penetration in obesity and with intestinal air impairing image quality. The flexibility ultrasonography offers the examiner also implies that a systematic approach during scanning is needed. This paper reviews the basic scanning techniques and new modalities such as contrast-enhanced ultrasound, elastography, strain rate imaging, hydrosonography, allergosonography, endoscopic sonography and nutritional imaging, and the literature on disease-specific findings in the small intestine. Some of these methods have shown clinical benefit, while others are under research and development to establish their role in the diagnostic repertoire. However, along with improved overall image quality of new ultrasound scanners, these methods have enabled more anatomical and physiological changes in the small intestine to be observed. Accordingly, ultrasound of the small intestine is an attractive clinical tool to study patients with a range of diseases.

  13. Small Signal Loudspeaker Impedance Emulator

    DEFF Research Database (Denmark)

    Iversen, Niels Elkjær; Knott, Arnold

    2014-01-01

    from driver to driver. Therefore, a loudspeaker emulator capable of adjusting its impedance to that of a given driver is desired for measurement purposes. This paper proposes a loudspeaker emulator circuit for small signals. Simulations and experimental results are compared and show that it is possible...

  14. Small firm transformation through IS

    NARCIS (Netherlands)

    Levy, Margi; Powell, Philip

    2008-01-01

    Globally, Small and Medium-sized Enterprises (SMEs) are encouraged, particularly by governments, to embrace c-business. Fully adopting e-business involves substantial change in firms, both internally and externally. However, there is little understanding of the mechanisms by which such business tran

  15. Small Turing universal signal machines

    Directory of Open Access Journals (Sweden)

    Jérôme Durand-Lose

    2009-06-01

    Full Text Available This article aims at providing signal machines as small as possible able to perform any computation (in the classical understanding. After presenting signal machines, it is shown how to get universal ones from Turing machines, cellular-automata and cyclic tag systems. Finally a halting universal signal machine with 13 meta-signals and 21 collision rules is presented.

  16. Microwave Levitation Of Small Objects

    Science.gov (United States)

    Watkins, John L.; Jackson, Henry W.

    1991-01-01

    Microwave radiation in resonant cavities used to levitate small objects, according to proposal. Feedback control and atmosphere not needed. Technique conceived for use in experiments on processing of materials in low gravitation of outer space, also used in normal Earth gravitation, albeit under some limitations.

  17. Small bowel transplantation: An overview

    NARCIS (Netherlands)

    R.W.F. de Bruin (Ron); E. Heineman (Erik); R.L. Marquet (Richard)

    1994-01-01

    textabstractSmall bowel transplantation (SBT) would, in theory, be the treatment of choice for patients suffering from the short bowel syndrome. Although SBT has been done with a considerable degree of success in some centers [36,145], it is by no means an established or widely applicable therapy fo

  18. Proceedings of Small Hydro 2009

    International Nuclear Information System (INIS)

    This conference was attended by speakers from government, industry and client organizations who addressed the growing interest in small hydro. Small hydro has been steadily increasing for years and will likely continue to increase as the supply of fossil fuels dwindles. The conference provided an opportunity to assess this renewable resources in terms of new estimates of micro (less than 100 kW), mini (100 kW to 1 MW) and small hydro (1 MW to 50 MW) facilities. Some presentations identified new locations for potential hydroelectric facilities and the development of projects at existing structures. New development mechanisms were reviewed, such as government funding initiatives, prospects for private funding, risk assessment and allocation. Local issues regarding small hydro development include quick routes to licensing and assessing and minimizing environmental impacts. The conference also provided an opportunity to highlight advances in technology to increase efficiency; refurbish and uprate existing, aging hydropower plants; and software developments. The conference featured 25 presentations, of which 10 have been catalogued separately for inclusion in this database. refs., tabs., figs

  19. Small rocket research and technology

    Science.gov (United States)

    Schneider, Steven; Biaglow, James

    1993-01-01

    Small chemical rockets are used on nearly all space missions. The small rocket program provides propulsion technology for civil and government space systems. Small rocket concepts are developed for systems which encompass reaction control for launch and orbit transfer systems, as well as on-board propulsion for large space systems and earth orbit and planetary spacecraft. Major roles for on-board propulsion include apogee kick, delta-V, de-orbit, drag makeup, final insertions, north-south stationkeeping, orbit change/trim, perigee kick, and reboost. The program encompasses efforts on earth-storable, space storable, and cryogenic propellants. The earth-storable propellants include nitrogen tetroxide (NTO) as an oxidizer with monomethylhydrazine (MMH) or anhydrous hydrazine (AH) as fuels. The space storable propellants include liquid oxygen (LOX) as an oxidizer with hydrazine or hydrocarbons such as liquid methane, ethane, and ethanol as fuels. Cryogenic propellants are LOX or gaseous oxygen (GOX) as oxidizers and liquid or gaseous hydrogen as fuels. Improved performance and lifetime for small chemical rockets are sought through the development of new predictive tools to understand the combustion and flow physics, the introduction of high temperature materials to eliminate fuel film cooling and its associated combustion inefficiency, and improved component designs to optimize performance. Improved predictive technology is sought through the comparison of both local and global predictions with experimental data. Results indicate that modeling of the injector and combustion process in small rockets needs improvement. High temperature materials require the development of fabrication processes, a durability data base in both laboratory and rocket environments, and basic engineering property data such as strength, creep, fatigue, and work hardening properties at both room and elevated temperature. Promising materials under development include iridium-coated rhenium and a

  20. Small is beautiful: models of small neuronal networks

    Science.gov (United States)

    Lamb, Damon G; Calabrese, Ronald L

    2013-01-01

    Modeling has contributed a great deal to our understanding of how individual neurons and neuronal networks function. In this review, we focus on models of the small neuronal networks of invertebrates, especially rhythmically active CPG networks. Models have elucidated many aspects of these networks, from identifying key interacting membrane properties to pointing out gaps in our understanding, for example missing neurons. Even the complex CPGs of vertebrates, such as those that underlie respiration, have been reduced to small network models to great effect. Modeling of these networks spans from simplified models, which are amenable to mathematical analyses, to very complicated biophysical models. Some researchers have now adopted a population approach, where they generate and analyze many related models that differ in a few to several judiciously chosen free parameters; often these parameters show variability across animals and thus justify the approach. Models of small neuronal networks will continue to expand and refine our understanding of how neuronal networks in all animals program motor output, process sensory information and learn. PMID:22364687

  1. Small Bodies, Big Discoveries: NASA's Small Bodies Education Program

    Science.gov (United States)

    Mayo, L.; Erickson, K. J.

    2014-12-01

    2014 is turning out to be a watershed year for celestial events involving the solar system's unsung heroes, small bodies. This includes the close flyby of comet C/2013 A1 / Siding Spring with Mars in October and the historic Rosetta mission with its Philae lander to comet 67P/Churyumov-Gerasimenko. Beyond 2014, the much anticipated 2015 Pluto flyby by New Horizons and the February Dawn Mission arrival at Ceres will take center stage. To deliver the excitement and wonder of our solar system's small bodies to worldwide audiences, NASA's JPL and GSFC education teams in partnership with NASA EDGE will reach out to the public through multiple venues including broadcast media, social media, science and math focused educational activities, observing challenges, interactive visualization tools like "Eyes on the Solar System" and more. This talk will highlight NASA's focused education effort to engage the public in small bodies mission science and the role these objects play in our understanding of the formation and evolution of the solar system.

  2. General Information about Small Cell Lung Cancer

    Science.gov (United States)

    ... Cancer Prevention Lung Cancer Screening Research Small Cell Lung Cancer Treatment (PDQ®)–Patient Version General Information About Small Cell Lung Cancer Go to Health Professional Version Key Points Small ...

  3. Stages of Small Cell Lung Cancer

    Science.gov (United States)

    ... Cancer Prevention Lung Cancer Screening Research Small Cell Lung Cancer Treatment (PDQ®)–Patient Version General Information About Small Cell Lung Cancer Go to Health Professional Version Key Points Small ...

  4. Treatment Option Overview (Small Cell Lung Cancer)

    Science.gov (United States)

    ... Cancer Prevention Lung Cancer Screening Research Small Cell Lung Cancer Treatment (PDQ®)–Patient Version General Information About Small Cell Lung Cancer Go to Health Professional Version Key Points Small ...

  5. C4 Photosynthetic Gene Expression in Light- and Dark-Grown Amaranth Cotyledons.

    Science.gov (United States)

    Wang, J. L.; Long, J. J.; Hotchkiss, T.; Berry, J. O.

    1993-08-01

    The patterns of expression for genes encoding several C4 photosynthetic enzymes were examined in light-grown and dark-grown (etiolated) cotyledons of amaranth (Amaranthus hypochondriacus), a dicotyledonous C4 plant. The large subunit and small subunit of ribulose-1,5-bisphosphate carboxylase (RuBPCase), phosphoenolpyruvate carboxylase (PEPCase), and pyruvate orthophosphate dikinase (PPdK) were all present in the cotyledons by d 2 after planting when the seedlings first emerged from the seed coat. Kranz anatomy was apparent in light-grown cotyledons throughout development, and the overall patterns of C4 gene expression were similar to those recently described for developing amaranth leaves (J.L. Wang, D.F. Klessig, J.O. Berry [1992] Plant Cell 4: 173-184). RuBPCase mRNA and proteins were present in both bundle sheath and mesophyll cells in a C3-like pattern during early development and became progressively more localized to bundle sheath cells in the C4-type pattern as the cotyledons expanded over 2 to 7 d. PEPCase and PPdK polypeptides were localized to mesophyll cells throughout development, even though PEPCase transcripts were detected in both bundle sheath and mesophyll cells. Kranz anatomy also developed in cotyledons grown in complete darkness. In 7-d-old dark-grown cotyledons, RuBPCase, PPdK, and PEPCase were all localized to the appropriate cell types, although at somewhat lower levels than in light-grown cotyledons. These findings demonstrate that the leaves and postembryonic cotyledons of amaranth undergo common developmental programs of C4 gene expression during maturation. Furthermore, light is not required for the cell-type-specific expression of genes encoding RuBPCase and other photosynthetic enzymes in this dicotyledonous C4 plant. PMID:12231890

  6. Structural analysis of CsoS1A and the protein shell of the Halothiobacillus neapolitanus carboxysome.

    Directory of Open Access Journals (Sweden)

    Yingssu Tsai

    2007-06-01

    Full Text Available The carboxysome is a bacterial organelle that functions to enhance the efficiency of CO2 fixation by encapsulating the enzymes ribulose bisphosphate carboxylase/oxygenase (RuBisCO and carbonic anhydrase. The outer shell of the carboxysome is reminiscent of a viral capsid, being constructed from many copies of a few small proteins. Here we describe the structure of the shell protein CsoS1A from the chemoautotrophic bacterium Halothiobacillus neapolitanus. The CsoS1A protein forms hexameric units that pack tightly together to form a molecular layer, which is perforated by narrow pores. Sulfate ions, soaked into crystals of CsoS1A, are observed in the pores of the molecular layer, supporting the idea that the pores could be the conduit for negatively charged metabolites such as bicarbonate, which must cross the shell. The problem of diffusion across a semiporous protein shell is discussed, with the conclusion that the shell is sufficiently porous to allow adequate transport of small molecules. The molecular layer formed by CsoS1A is similar to the recently observed layers formed by cyanobacterial carboxysome shell proteins. This similarity supports the argument that the layers observed represent the natural structure of the facets of the carboxysome shell. Insights into carboxysome function are provided by comparisons of the carboxysome shell to viral capsids, and a comparison of its pores to the pores of transmembrane protein channels.

  7. Propulsion Challenges for Small Spacecraft: 2005

    Institute of Scientific and Technical Information of China (English)

    Vadim Zakirov; LI Luming

    2006-01-01

    Small (<100 kg) spacecrafts are being developed in many countries but their propulsion systems still have many challenges. Although there is demand for small spacecraft propulsion, the number of missions at present is small due to several commercial and technical reasons. Poor performance of existing small spacecraft propulsion systems is one of the main reasons for the small number of missions. Several reasons are given for the poor performance of existing small spacecraft propulsion. Suggested improvements focus on small spacecraft and propulsion hardware mass optimization rather than on specific impulse enhancement. Propellantless propulsion systems are also recommended for small spacecraft interplanetary missions.

  8. Going global - growing small businesses

    International Nuclear Information System (INIS)

    The Going Global Energy Steering Committee was established to help small and medium-sized Canadian enterprises to compete in the electrical power industry in the Asia-Pacific region primarily, but also in Eastern Europe and Latin America. The aim is to provide market intelligence, and help with forming consortia for financing. A small to medium-sized business can be defined as one with 50 to 500 employees. Big businesses no longer have the same competitive advantages that they once had, because automated systems can make short production runs just as cost-effective as long ones, and because computerization, automation and rising productivity mean that fewer workers are required than formerly

  9. Trust Propagation in Small Worlds

    DEFF Research Database (Denmark)

    Gray, Elizabeth; Seigneur, Jean-Marc; Chen, Yong;

    2003-01-01

    do not scale well. We aim to develop trust-based security mechanisms using small world concepts to optimise formation and propagation of trust amongst entities in these vast networks. In this regard, we surmise that in a very large mobile ad hoc network, trust, risk, and recommendations can be...... propagated through relatively short paths connecting entities. Our work describes the design of trust-formation and risk-assessment systems, as well as that of an entity recognition scheme, within the context of the small world network topology.......The possibility of a massive, networked infrastructure of diverse entities partaking in collaborative applications with each other increases more and more with the proliferation of mobile devices and the development of ad hoc networking technologies. In this context, traditional security measures...

  10. Remotely sensed small reservoir monitoring

    Science.gov (United States)

    Eilander, Dirk; Annor, Frank; Iannini, Lorenzo; van de Giesen, Nick

    2013-04-01

    A new 'growing' maximum likelihood classification algorithm for small reservoir delineation has been developed and is tested with Radarsat-2 data for reservoirs in the semi-arid Upper East Region, Ghana. The delineation algorithm is able to find the land-water boundary from SAR imagery for different weather and environmental conditions. As such, the algorithm allows for remote sensed operational monitoring of small reservoirs. Multipurpose small reservoirs (1-100 ha) are important for many livelihoods in rural semi-arid West Africa. In order to manage and plan these reservoirs and to assess their hydrological impact at a river basin scale, it is important to monitor their water storage fluctuation. Several studies on remotely sensed reservoir mapping have recently been published, but no single method yields good results for all weather and environmental conditions. Detection of small reservoirs from optical satellite imagery using supervised maximum likelihood classification is a well proved method. The application of this method for the monitoring of small reservoirs is however limited because of its dependence on cloud-free day-acquisitions. Delineation from SAR images is promising, but because of difficulties with wind induced Bragg-scattering and low contrast between the water surface and the dried-out surroundings at the end of the dry season, only quasi manual methods have been applied successfully. A smart combination of optical satellite based detection combined with a delineation method for SAR imagery is proposed. From the optical satellite based small reservoir detection the reservoir window is determined in which the 'growing' maximum likelihood classification on SAR images is performed. A water-class seed and land-class seed are implemented and grown dependent on the likelihood of a pixel to belong to one class. The likelihood is calculated based on the probability distributions of the growing land and water populations. Combinations of single

  11. Unveiling small sphere's scattering behavior

    CERN Document Server

    Tzarouchis, Dimitrios C; Sihvola, Ari

    2016-01-01

    A classical way for exploring the scattering behavior of a small sphere is to approximate Mie coefficients with a Taylor series expansion. This ansatz delivered a plethora of insightful results, mostly for small spheres supporting electric localized plasmonic resonances. However, many scattering aspects are still uncharted, especially for the case of magnetic resonances. Here, an alternative system ansatz is proposed based on the Pad\\'e approximants for the Mie coefficients. The extracted results reveal new aspects, such as the existence of a self-regulating radiative damping mechanism for the first magnetic resonance. Hence, a systematic way of exploring the scattering behavior is introduced, sharpening our understanding about sphere's scattering behavior and its emergent functionalities.

  12. Knowledge Management in Small Firms

    Science.gov (United States)

    Panyasorn, Jessada; Panteli, Niki; Powell, Philip

    This paper explores knowledge management in small and medium-sized firms (SMEs). It investigates the use of Lotus Notes in SMEs of a developing country as a counterpoint to the large firm, developed country emphasis of existing research. It develops taxonomy of Lotus Notes use within the context of different knowledge management processes; notably communicating, co-ordinating and collaborating. The study employs an interpretive approach using three case studies. The key findings suggest that publishing, searching, sharing and retrieving are the user modes for enabling sharing and storing information. Evidence of knowledge creation is found at the departmental level but not at the organizational level. Further, small firms may explore more groupware potential than large organizations and this reflects their different context. Finally, implications for further research are identified.

  13. [Sarcomas of the small intestine].

    Science.gov (United States)

    Beyrouti, M L; Abid, M; Beyrouti, R; Ben Amar, M; Gargouri, F; Frikha, F; Affes, N; Boujelbene, S; Ghorbel, A

    2005-03-12

    Sarcomas of the small intestine are rare, clearly differentiated, malignant, mesenchymatous tumours that can be of smooth muscle, Schwann cell or fibroblastic origin. From a clinical point of view, the pain and abdominal mass are the 2 types of symptoms that frequently reveal the disease. In rare cases, sarcomas of the small intestine are manifested by an acute complication. No imaging method can clearly confirm the diagnosis. Before immunohistochemistry, differential diagnosis was made on undifferentiated mesenchymatous "stromal" tumours, which are also rare. Exeresis must be complete and without perforation of the tumour because of the risk of locoregional relapse. The benefits provided by chemotherapy and radiotherapy are limited because of the low mitotic activity of the tumour cells and its weak vascularisation. Long-term survival is limited by poor prognosis criteria: high grade malignancy, size greater than 5 cm, tumour extension, perforation of the tumour, quality of surgical resection and histological type. PMID:15859576

  14. Parton distributions at small x

    International Nuclear Information System (INIS)

    We perform a next-to-leading order quantum chromodynamics (QCD) analysis of the recent data for deep-inelastic lepton-nucleon scattering and related processes, in which we pay particular attention to the forms of the parton distributions at very small x. We discuss in detail, and we incorporate in the analysis, the theoretical QCD results leading to the singular x-1/2 type of behaviour of the gluon and sea quark distributions, as well as the modifications due to shadowing effects. We find the QCD shadowing corrections are significant for x ''approx -3 even though the parton distributions are below their saturation limit. We give predictions for the structure functions F2 and FL accessible at HERA, and for W and Z productions up to linear hard core pinch devices (LHC) and superconducting supercollider (SSC) energies. We discuss the possibility of experiments at these colliders probing the parton distributions in the very small x region. (author)

  15. Unitarity at small Bjorken x

    CERN Document Server

    Weigert, H

    2002-01-01

    This paper presents a solution to the nonlinear small x ``projectile side'' evolution equations as derived by Balitskii in 1996. The solution is based on functional Fokker-Planck methods. The fixed point at small x is explicitly calculated and all correlation functions in this limit are determined. They show clear saturation and unitarization properties. Scaling laws that hold during the saturation phase and throughout the whole course of the evolution are established. The corresponding Langevin equations are given as a basis for numerical simulations opening the field for future studies of dynamical issues of the evolution not analytically accessible. The methods used may be extended to the ``target side'' equations of Jalilian-Marian, Kovner, Leonidov and Weigert.

  16. Unitarity at small Bjorken x

    International Nuclear Information System (INIS)

    This paper presents a solution to the nonlinear small-x 'projectile side' evolution equations as derived by Balitskii in 1996. The solution is based on functional Fokker-Planck methods. The fixed point at small x is explicitly calculated and all correlation functions in this limit are determined. They show clear saturation and unitarization properties. Scaling laws that hold during the saturation phase and throughout the whole course of the evolution are established. The corresponding Langevin equations are given as a basis for numerical simulations opening the field for future studies of dynamical issues of the evolution not analytically accessible. The methods used may be extended to the 'target side' equations of Jalilian-Marian, Kovner, Leonidov and Weigert

  17. Unitarity at small Bjorken x

    Energy Technology Data Exchange (ETDEWEB)

    Weigert, Heribert E-mail: heribert.weigert@physik.uni-regensburg.de

    2002-06-03

    This paper presents a solution to the nonlinear small-x 'projectile side' evolution equations as derived by Balitskii in 1996. The solution is based on functional Fokker-Planck methods. The fixed point at small x is explicitly calculated and all correlation functions in this limit are determined. They show clear saturation and unitarization properties. Scaling laws that hold during the saturation phase and throughout the whole course of the evolution are established. The corresponding Langevin equations are given as a basis for numerical simulations opening the field for future studies of dynamical issues of the evolution not analytically accessible. The methods used may be extended to the 'target side' equations of Jalilian-Marian, Kovner, Leonidov and Weigert.

  18. Small black holes on cylinders

    International Nuclear Information System (INIS)

    We find the metric of small black holes on cylinders, i.e. neutral and static black holes with a small mass in d-dimensional Minkowski space times a circle. The metric is found using an ansatz for black holes on cylinders proposed in J. High Energy Phys. 05, 032 (2002). We use the new metric to compute corrections to the thermodynamics which is seen to deviate from that of the (d+1)-dimensional Schwarzschild black hole. Moreover, we compute the leading correction to the relative binding energy which is found to be non-zero. We discuss the consequences of these results for the general understanding of black holes and we connect the results to the phase structure of black holes and strings on cylinders

  19. Neutronic design of small reactors

    International Nuclear Information System (INIS)

    Small reactors design is one of the main activities of AREVA TA. At the time, AREVA TA main projects are oriented towards research reactors and reactors for military naval propulsion. Due to differences in the physics and performances to meet, each kind of small reactor leads to specific modelling needs. Many computing tools have been developed in order to successfully carry out these projects. These schemes are mainly based on the use of TRIPOLI, MCNP, APOLLO2 and CRONOS2 codes. In that framework, a multi-purpose pre/post processing tool named CHARM is being developed by AREVA NP in partnership with AREVA TA in order to integrate small reactors specification. CHARM is used to elaborate APOLLO2 input data while various dedicated tools are used to automatically generate TRIPOLI and MNCP input data. These 3D numerical models need a very accurate spatial description to perform specific calculations. As an example, for the JHR design, after calculating 3D burn up by APOLLO2/MOC models, the data is fed back into a TRIPOLI model used for safety analyses. This paper presents our methodology for the small core design and 3 examples: 1) The calculation scheme for the JHR (Jules Horowitz Reactor) neutronic studies. These design studies are a recent illustration of combined use of both deterministic and probabilistic codes, 2) The use of CHARM, with the modelling of a JHR core. The purpose of CHARM- V2, based on Open Cascade Technology, is to provide a pre/post processing tool for APOLLO2/MOC, TRIPOLI4 and MCNP solvers, 3) The depletion Monte Carlo calculation of a MTR core. (author)

  20. Doing Astronomy with Small Telescopes

    OpenAIRE

    Kangujam Yugindro Singh; Irom Ablu Meitei; Salam Ajitkumar Singh

    2014-01-01

    We are playing a lead role for growth of astronomy and its quality teaching and research in Manipur, a State located at northeast India (longitude = 93°58'E; latitude = 24°44'N; altitude = 782 m). We have innovatively designed and constructed three cost effective observatories, each costing a few hundred USD. These observatories are completely different in design and are perfectly usable for doing serious work on astronomical observation and measurements, using small ground-...

  1. Metagenomic small molecule discovery methods

    OpenAIRE

    Charlop-Powers, Zachary; Milshteyn, Aleksandr; Brady, Sean F

    2014-01-01

    Metagenomic approaches to natural product discovery provide the means of harvesting bioactive small molecules synthesized by environmental bacteria without the requirement of first culturing these organisms. Advances in sequencing technologies and general metagenomic methods are beginning to provide the tools necessary to unlock the unexplored biosynthetic potential encoded by the genomes of uncultured environmental bacteria. Here, we highlight recent advances in sequence- and functional- bas...

  2. Nucleation barrier in small volumes

    Czech Academy of Sciences Publication Activity Database

    Kožíšek, Zdeněk; Demo, Pavel; Sveshnikov, Alexey; Tichá, Petra

    Bratislava: N, 2009 - (Koman, M.; Mikloš, D.), s. 36-37 ISBN 978-80-89088-81-2. [Development of Materials Science in Research and Education - DMS -RE 2009 /19./. Závažná Poruba (SK), 31.08.2009-04.09.2009] R&D Projects: GA AV ČR IAA100100806 Institutional research plan: CEZ:AV0Z10100521 Keywords : nucleation * phase transition in small volumes Subject RIV: BM - Solid Matter Physics ; Magnetism

  3. Design of small table saws

    OpenAIRE

    Stuchlík, Petr

    2016-01-01

    The main aim of this thesis is the engineering design of a small table circular saw according to the specified parameters. At first, the types of machines available on the market are named and analyzed in detail. Afterwards, the engineering design of the machine is created. The following part is concerned with the individual parts,, tilting of the saw blade, maximum cutting pull through linkage with motion screw and checking of deformation of he critical parts of the machine. The previously s...

  4. Small gauge vitrectomy: Recent update

    OpenAIRE

    Sumeet Khanduja; Ashish Kakkar; Saptrishi Majumdar; Rajpal Vohra; Satpal Garg

    2013-01-01

    Small gauge vitrectomy, also known as minimally invasive vitreous surgery (MIVS), is a classic example of progress in biomedical engineering. Disparity in conjunctival and scleral wound location and reduction in wound diameter are its core principles. Fluidic changes include increased pressure head loss with consequent reduction in infusional flow rate and use of higher aspiration vacuum at the cutter port. Increase An increase in port open/port closed time maintains an adequate rate of vitre...

  5. Small intestinal bacterial overgrowth syndrome

    OpenAIRE

    Jan Bures, Jiri Cyrany, Darina Kohoutova, Miroslav Förstl, Stanislav Rejchrt, Jaroslav Kvetina, Viktor Vorisek, Marcela Kopacova

    2010-01-01

    Human intestinal microbiota create a complex polymicrobial ecology. This is characterised by its high population density, wide diversity and complexity of interaction. Any dysbalance of this complex intestinal microbiome, both qualitative and quantitative, might have serious health consequence for a macro-organism, including small intestinal bacterial overgrowth syndrome (SIBO). SIBO is defined as an increase in the number and/or alteration in the type of bacteria in the upper gastrointestina...

  6. Small intestinal bacterial overgrowth syndrome.

    OpenAIRE

    Bures, J.; Cyrany, J.; Kohoutova, D.; Förstl, M.; Rejchrt, S.; Kvetina, J.; Vorisek, V.; Kopacova, M.

    2010-01-01

    Human intestinal microbiota create a complex polymicrobial ecology. This is characterised by its high population density, wide diversity and complexity of interaction. Any dysbalance of this complex intestinal microbiome, both qualitative and quantitative, might have serious health consequence for a macro-organism, including small intestinal bacterial overgrowth syndrome (SIBO). SIBO is defined as an increase in the number and/or alteration in the type of bacteria in the upper gastrointestina...

  7. The small world of psychopathology

    OpenAIRE

    Denny Borsboom; Cramer, Angélique O. J.; Schmittmann, Verena D.; Sacha Epskamp; Waldorp, Lourens J.

    2011-01-01

    BACKGROUND: Mental disorders are highly comorbid: people having one disorder are likely to have another as well. We explain empirical comorbidity patterns based on a network model of psychiatric symptoms, derived from an analysis of symptom overlap in the Diagnostic and Statistical Manual of Mental Disorders-IV (DSM-IV). PRINCIPAL FINDINGS: We show that a) half of the symptoms in the DSM-IV network are connected, b) the architecture of these connections conforms to a small world structure, fe...

  8. The small world of psychopathology

    OpenAIRE

    Borsboom, D; Cramer, A.O.J.; Schmittmann, V.D.; Epskamp, S.; Waldorp, L.J.

    2011-01-01

    Background Mental disorders are highly comorbid: people having one disorder are likely to have another as well. We explain empirical comorbidity patterns based on a network model of psychiatric symptoms, derived from an analysis of symptom overlap in the Diagnostic and Statistical Manual of Mental Disorders-IV (DSM-IV). Principal Findings We show that a) half of the symptoms in the DSM-IV network are connected, b) the architecture of these connections conforms to a small world structure, feat...

  9. Rhapsody on small processor platforms

    OpenAIRE

    Andersson, Per-Oskar

    2008-01-01

    Rhapsody is a Model-Driven Development (MDD) tool for embedded and real-time system design. The purpose of this thesis is to determine if Rhapsody can be used for software development on small processor platforms such as the Atmel AVR. Rhapsody is normally used on platforms running an operating system. Therefore certain adaptations are needed in order to use it on platforms without an operating system. These adaptations and their affect on the usability of the tool, advantages and disadvantag...

  10. Twisted Yangians of small rank

    Science.gov (United States)

    Guay, Nicolas; Regelskis, Vidas; Wendlandt, Curtis

    2016-04-01

    We study quantized enveloping algebras called twisted Yangians associated with the symmetric pairs of types CI, BDI, and DIII (in Cartan's classification) when the rank is small. We establish isomorphisms between these twisted Yangians and the well known Olshanskii's twisted Yangians of types AI and AII, and also with the Molev-Ragoucy reflection algebras associated with symmetric pairs of type AIII. We also construct isomorphisms with twisted Yangians in Drinfeld's original presentation.

  11. Innovation in small food firms

    OpenAIRE

    Avermaete, Tessa

    2012-01-01

    Internationalisation of the world food market, structural changes in the food chain, changing consumption patterns and the strengthening of food regulation lie on the basis of the fast developments in the food and drink industry. Facing increased competition, small food firms are generally limited in exploiting economies of scale. Alternatively, they are more likely to seek for strategies that are based on the introduction of new products, optimising processing, the reorganisation of manageme...

  12. Small Business Innovations (Robotic Wrist)

    Science.gov (United States)

    1991-01-01

    Under a Langley Research Center Small Business Innovation Research (SBIR) contract, Ross-Hime Designs, Inc. Minneapolis, MN, developed the Omni-Wrist actuator, which has a 25-pound capacity, 180 degrees of pitch/yaw, and 360 degrees of roll. Company literature calls it "the first successful singularity-free high-precision (robotic) wrist." Applications include spray painting, sealing, ultrasonic testing, welding and a variety of nuclear industry, aerospace and military uses.

  13. Small Business Innovations (Integrated Database)

    Science.gov (United States)

    1992-01-01

    Because of the diversity of NASA's information systems, it was necessary to develop DAVID as a central database management system. Under a Small Business Innovation Research (SBIR) grant, Ken Wanderman and Associates, Inc. designed software tools enabling scientists to interface with DAVID and commercial database management systems, as well as artificial intelligence programs. The software has been installed at a number of data centers and is commercially available.

  14. Small Business Innovations (Fiber Optics)

    Science.gov (United States)

    1991-01-01

    Foster-Miller, Inc. Waltham, MA developed the In-Situ Fiber Optic Polymer Reaction Monitor which could lead to higher yields and lower costs in complex composite manufacturing. The monitor, developed under a Small Business Innovation Research (SBIR) contract with Langley Research Center, uses an infrared, fiber optic sensor to track the molecular vibrational characteristics of a composite part while it is being cured. It is the first analytical system capable of directly measuring the chemistry of advanced composite materials.

  15. Vibration Testing for Small Satellites

    OpenAIRE

    Wilson, Delbert

    1989-01-01

    Most people involved in the design and construction of small satellites are unfamiliar with vibration testing. Yet most satellites undergo vibration testing to qualify them for flight. Some familiarity with the basic aspects of vibration testing is needed to insure that a vibration test on a satellite is valid. This paper sets forth the basic equipment, practices and concepts of vibration testing. It provides guidelines for specifying a vibration test, designing fixtures, attaching instrument...

  16. Implantable telemetry for small animals

    Science.gov (United States)

    1982-03-01

    A series of totally implantable telemetry devices for use in measuring deep body parameters in small animals were developed. Under a collaborative agreement with NASA, several of these systems; the continuous wave Doppler ultrasonic flowmeter, the multichannel telemetry system, and the inductively-powered dual channel cardiac pacer were evaluated in a series of ten mongrel dogs (15 to 20 kg.). These systems were used to measure ascending aortic and coronary blood flow, aortic pressure, and subcutaneous EKG.

  17. The evolution of small change

    OpenAIRE

    Thomas J. Sargent; Francois R. Velde

    1997-01-01

    Western Europe was plagued with currency shortages from the 14th to the 19th century, at which time a `standard formula' had been devised to cure the problem. We document the evolution of mon- etary theory, policy experiments and minting tech- nology over the course of six hundred years. In a companion paper, we use a cash-in-advance model of commodity money to provide an analytical frame- work for the problem of small change.

  18. Small intestinal bacterial overgrowth syndrome

    Institute of Scientific and Technical Information of China (English)

    Jan; Bures; Jiri; Cyrany; Darina; Kohoutova; Miroslav; Frstl; Stanislav; Rejchrt; Jaroslav; Kvetina; Viktor; Vorisek; Marcela; Kopacova

    2010-01-01

    Human intestinal microbiota create a complex polymi-crobial ecology. This is characterised by its high population density, wide diversity and complexity of interaction. Any dysbalance of this complex intestinal microbiome, both qualitative and quantitative, might have serious health consequence for a macro-organism, including small intestinal bacterial overgrowth syndrome (SIBO).SIBO is defined as an increase in the number and/or alteration in the type of bacteria in the upper gastro-intestinal tract. There...

  19. Evolution of small prokaryotic genomes

    Directory of Open Access Journals (Sweden)

    David José Martínez-Cano

    2015-01-01

    Full Text Available As revealed by genome sequencing, the biology of prokaryotes with reduced genomes is strikingly diverse. These include free-living prokaryotes with ~800 genes as well as endosymbiotic bacteria with as few as ~140 genes. Comparative genomics is revealing the evolutionary mechanisms that led to these small genomes. In the case of free-living prokaryotes, natural selection directly favored genome reduction, while in the case of endosymbiotic prokaryotes neutral processes played a more prominent role. However, new experimental data suggest that selective processes may be at operation as well for endosymbiotic prokaryotes at least during the first stages of genome reduction. Endosymbiotic prokaryotes have evolved diverse strategies for living with reduced gene sets inside a host-defined medium. These include utilization of host-encoded functions (some of them coded by genes acquired by gene transfer from the endosymbiont and/or other bacteria; metabolic complementation between co-symbionts; and forming consortiums with other bacteria within the host. Recent genome sequencing projects of intracellular mutualistic bacteria showed that previously believed universal evolutionary trends like reduced G+C content and conservation of genome synteny are not always present in highly reduced genomes. Finally, the simplified molecular machinery of some of these organisms with small genomes may be used to aid in the design of artificial minimal cells. Here we review recent genomic discoveries of the biology of prokaryotes endowed with small gene sets and discuss the evolutionary mechanisms that have been proposed to explain their peculiar nature.

  20. Superconductivity of small metal grains

    Institute of Scientific and Technical Information of China (English)

    ZHENG; Renrong; CHEN; Zhiqian; ZHU; Shunquan

    2005-01-01

    The formulas of the energy gap and superconducting critical temperature appropriate for systems with both odd and even number of electrons are derived; the bases of the derivations are BCS theory and energy level statistics. Numerical results qualitatively agree with the experimental phenomena. i.e., the superconductivity of small metallic grains will first enhance then decrease to zero when the grain are getting smaller and smaller. The calculations indicate that the above phenomena happen in the metallic grains belonging to Gaussian Orthogonal Ensemble (GOE) and Gaussian Unitary ensemble (GUE) with zero spin; The superconductivity of small metallic grains in Gaussian Symplectic Ensemble (GSE) will monotonically decrease to zero with the decreasing of the grain size. The analyses suggest that the superconductivity enhancements come from pairing and the balance of the strengths between spin-orbital coupling and external magnetic field. In order to take the latter into account, it is necessary to include the level statistics given by Random Matrix Theory (RMT) in describing small metallic grains.

  1. Small Break Air Ingress Experiment

    Energy Technology Data Exchange (ETDEWEB)

    Chang Oh; Eung Soo Kim

    2011-09-01

    The small break air-ingress experiment, described in this report, is designed to investigate air-ingress phenomena postulated to occur in pipes in a very high temperature gas-cooled reactor (VHTRs). During this experiment, air-ingress rates were measured for various flow and break conditions through small holes drilled into a pipe of the experimental apparatus. The holes were drilled at right angles to the pipe wall such that a direction vector drawn from the pipe centerline to the center of each hole was at right angles with respect to the pipe centerline. Thus the orientation of each hole was obtained by measuring the included angle between the direction vector of each hole with respect to a reference line anchored on the pipe centerline and pointing in the direction of the gravitational force. Using this reference system, the influence of several important parameters on the air ingress flow rate were measured including break orientation, break size, and flow velocity . The approach used to study the influence of these parameters on air ingress is based on measuring the changes in oxygen concentrations at various locations in the helium flow circulation system as a function of time using oxygen sensors (or detectors) to estimate the air-ingress rates through the holes. The test-section is constructed of a stainless steel pipe which had small holes drilled at the desired locations.

  2. Chronic Hemodialysis in Small Children.

    Science.gov (United States)

    Novljan, Gregor; Rus, Rina R; Premru, Vladimir; Ponikvar, Rafael; Battelino, Nina

    2016-06-01

    When peritoneal dialysis is inapplicable, chronic hemodialysis (HD) becomes the only available treatment option in small children. Due to small patient size, central venous catheters (CVC) are mainly used for vascular access. Over the past 4 years, four children weighing less than 15 kg received chronic HD in our unit. A total of 848 dialysis sessions were performed. Altogether, 21 catheters were inserted. In all but one occasion, uncuffed catheters were used. Catheter revision was performed 15 times during the study period, either due to infection or catheter malfunction. The median number of catheter revisions and the median line survival was 3.0/patient-year and 53 days (range; 6-373 days), respectively. There were 14 episodes of catheter related infections requiring 11 CVC revisions (78.6%). The median rate of line infections was 2.8/patient-year. Chronic HD in small children is demanding and labor intensive. Issues pertain mainly to CVCs and limit its long-term use. PMID:27312919

  3. Study of the properties of Ribulose 1,5-biphosphate carboxylase/oxygenase from maize (Zea mays) and wheat (Triticum aestivum) by incorporation of 14{sub C}O2; Estudio de las propiedades de la Ribulosa-1,5-Difosfato Carboxilasa/Oxigenasa de maiz (Zea Mais) y de trigo (Triticum Aestivum), por incorporacion de CO{sub 2} marcado con 14{sub C}O2

    Energy Technology Data Exchange (ETDEWEB)

    Garcia, M. D.; Saez, R. M.

    1982-07-01

    After a bibliographic review of the properties of RuBP-carboxylase/oxygenase, a methodology is described which allows the treatment of a large number of samples for the assay of the enzyme activity. 14{sup C}O{sub 3}HNa is used as a marker for the counting of the incorporated radioactivity as acid insoluble material. 14''CC{sub 2} from the labeled sodium bicarbonate is the species used by the enzyme both as an activator as well as a substrate. The following experiments are described and its results given: Determination of the optimal conditions for the activation of the enzyme; study of the kinetics of the catalytic action; effect of the Mg{sup 2} concentration and determination of the Km{sub (s)} from CO{sub 2} and ribulose 1,5-biphosphate; also determination of the optimum pH at different concentrations of CO{sub 2}2 and Mg{sup 2}. (Author) 64 refs.

  4. 76 FR 61626 - Small Business Subcontracting

    Science.gov (United States)

    2011-10-05

    ... ADMINISTRATION 13 CFR Parts 121 and 125 RIN 3245-AG22 Small Business Subcontracting AGENCY: U.S. Small Business Administration. ACTION: Proposed rule. SUMMARY: The U.S. Small Business Administration (SBA or Agency) is... submissions: Dean Koppel, U.S. Small Business Administration, Office of Government Contracting, 409...

  5. Small satellite solar array substrate

    Science.gov (United States)

    Fiore, John N.; Rosanova, Giulio

    1994-01-01

    The SMall EXplorer (SMEX) Fast Auroral SnapshoT (FAST) spacecraft was developed to investigate plasma physics of auroral phenomena at high orbital altitude. The FAST satellite comprises a variety of deployable booms with sensors on the ends, and instruments that protrude from the main body of the spacecraft to obtain the plasma and electromagnetic fields data. This required the plasma disturbance around the satellite to be kept to a minimum. A non deployable, body mounted solar array was implemented. This led to the design of a light weight solar array substrate with a high degree of structural integrity.

  6. Emissions from small heating plants

    International Nuclear Information System (INIS)

    This study treats emissions from and economy of small biofuelled heating plants. In order to reduce emissions of particulates below 100 mg/nm3, the plants must be equipped with flue gas cleaning systems superior to cyclones/multicyclones. The use of pellets or pulverized fuel does not necessarily give emissions lower than 100 mg/nm3. However, the study shows that the CO-emissions can be kept low. An economic analysis for biofuelled plants is also presented 18 tabs, 18 figs

  7. GEF small grants programme - overview

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-12-01

    This paper describes the GEF small grants program which seeks to enhance the role of households and communities in conserving global biodiversity, mitigating global climate change, and protecting international waters. Grants up to $50k have been granted for projects in 33 countries, with plans for 12 other countries. The author describes the framework that the program works under, and the methodology followed in developing and planning projects. The approach to climate change concerns is to emphasize the development of non-carbon energy development activities to provide energy sources and economic development.

  8. Valorization of small uranium deposits

    International Nuclear Information System (INIS)

    For the valorization of small uranium deposits, the Total Mining Co. has developed an original leaching process in immersed live stalls, allied to the treatment of uraniferous liquors on ion exchanger resins. This technology was put in operation at Saint-Pierre (Cantal) in 1976, at La Ribiere (Creuse) in 1982 and in Bertholene (Aveyron) in 1985. In the last mentioned installation, 75 t of uranium can be produced per year on the basis of 60 000 t of ore containing 1.5 kg of uranium per ton. The investments per kg of uranium have thus been reduced to a quarter of their value in a conventional plant

  9. Small angle scattering and polymers

    International Nuclear Information System (INIS)

    The determination of polymer structure is a problem of interest for both statistical physics and industrial applications. The average polymer structure is defined. Then, it is shown why small angle scattering, associated with isotopic substitution, is very well suited to the measurement of the chain conformation. The corresponding example is the old, but pedagogic, measurement of the chain form factor in the polymer melt. The powerful contrast variation method is illustrated by a recent determination of the concentration profile of a polymer interface. (author) 12 figs., 48 refs

  10. Small helical flux compression amplifiers

    International Nuclear Information System (INIS)

    Small, explosively compressed, magnetic flux transducers with many closely spaced helical turns are investigated theoretically and experimentally. The analysis is limited to linear operation, but takes into account load influence, proximity effects, and switching delays. The latter are due to retarded breakdown in the wire insulation and to the finite decay time of the magnetic field in the wire. More than 150 experiments showed considerable data scatter. Shots which exhibited low clocking and high amplification were in good agreement with the theory. The main conclusion is that device performance is limited not only by flux loss, but by flux remaining in the generator after compression

  11. Small angle scattering and polymers

    Energy Technology Data Exchange (ETDEWEB)

    Cotton, J.P. [Laboratoire Leon Brillouin (LLB) - Centre d`Etudes de Saclay, 91 - Gif-sur-Yvette (France)

    1996-12-31

    The determination of polymer structure is a problem of interest for both statistical physics and industrial applications. The average polymer structure is defined. Then, it is shown why small angle scattering, associated with isotopic substitution, is very well suited to the measurement of the chain conformation. The corresponding example is the old, but pedagogic, measurement of the chain form factor in the polymer melt. The powerful contrast variation method is illustrated by a recent determination of the concentration profile of a polymer interface. (author) 12 figs., 48 refs.

  12. Minimum Q Electrically Small Antennas

    DEFF Research Database (Denmark)

    Kim, O. S.

    2012-01-01

    Theoretically, the minimum radiation quality factor Q of an isolated resonance can be achieved in a spherical electrically small antenna by combining TM1m and TE1m spherical modes, provided that the stored energy in the antenna spherical volume is totally suppressed. Using closed-form expressions...... results for a multiarm spherical helix antenna confirm the theoretical predictions. For example, a 4-arm spherical helix antenna with a magnetic-coated perfectly electrically conducting core (ka=0.254) exhibits the Q of 0.66 times the Chu lower bound, or 1.25 times the minimum Q....

  13. Small signal microwave amplifier design

    CERN Document Server

    Grosch, Theodore

    2000-01-01

    This book explains techniques and examples for designing stable amplifiers for high-frequency applications in which the signal is small and the amplifier circuit is linear. An in-depth discussion of linear network theory provides the foundation needed to develop actual designs. Examples throughout the book will show you how to apply the knowledge gained in each chapter leading to the complex design of low noise amplifiers. Many exercises at the end of each chapter will help students to practice their skills. The solutions to these design problems are available in an accompanying solutions book

  14. 高脂饮食对老年大鼠骨骼肌脂肪酸含量及乙酰辅酶A羧化酶表达和活性的影响%Effects of high-fat diet on fatty acid metabolism, expression and activity of acetyl-coenzyme A carboxylase in skeletal muscle in aged rats

    Institute of Scientific and Technical Information of China (English)

    胡淑国; 宋光耀; 王敬; 高宇

    2010-01-01

    目的 探讨增龄和高脂饮食对大鼠骨骼肌脂肪酸含量及乙酰辅酶A羧化酶(acetyl-coenzyme A carboxylase,ACC)表达和活性的影响.方法 将22~24月龄雄性Wistar大鼠随机分为老年对照组和高脂组;4~5月龄大鼠作为青年对照组.老年对照组和青年对照组给基础饲料,高脂组给予高脂饲料,喂养8周.用高胰岛素-正葡萄糖钳夹实验评价各组大鼠胰岛素敏感性,用全自动生化分析仪测定骨骼肌三酰甘油,用荧光分光光度计测定骨骼肌总的长链脂酰辅酶A含量,用Western-blot方法测定骨骼肌ACC、和磷酸化ACC(P-ACC)蛋白表达.结果 (1)老年对照组空腹血糖、胰岛索和游离脂肪酸高于青年对照组,高脂组上述几项指标进一步升高,并且出现血清三酰甘油和总胆固醇水平增高;(2)老年对照组葡萄糖输注率(glucose infusion rates,GIR)低于青年对照组,高脂组GIR低于老年对照组,高脂组GIR在8周末低于4周末;(3)老年对照组骨骼肌三酰甘油及长链脂酰辅酶A含量高于青年对照组,高脂组与老年对照组比较进一步升高;(4)老年对照组与青年对照组之间、高脂组与老年对照组之间骨骼肌ACC蛋白表达均无明显变化(P>0.05);骨骼肌P-ACC蛋白水平在老年对照组低于青年对照组,高脂组与老年对照组比较进一步降低(P0.05). The protein levels of P-ACC in skeletal muscle were lower in OC group, and much lower in HF group than in YC group (P<0.05 or P<0.01). Conclusions Compared with young rats, abnormal fatty acid metabolism and insulin resistance always exist in aged rats. High-fat feeding results in a significant increase in lipid content in skeletal muscle. Alterations of ACC activity may contribute to fat accumulation in skeletal muscle and insulin resistance.

  15. Crescimento e competitividade de biótipos de capim-colchão resistente e suscetível aos herbicidas inibidores da acetil coenzima A carboxilase Growth and competitiveness of biotypes of crabgrass resistant and susceptible to acetyl coenzyme A carboxylase inhibiting herbicides

    Directory of Open Access Journals (Sweden)

    Ramiro Fernando López Ovejero

    2007-01-01

    Full Text Available O objetivo deste trabalho foi comparar o crescimento e a habilidade competitiva de dois biótipos de capim-colchão (Digitaria ciliaris, um resistente (R e outro suscetível (S aos herbicidas inibidores da acetil coenzima A carboxilase. O crescimento dos biótipos foi determinado pela coleta da matéria seca das plantas, aos 14, 21, 25, 28, 34, 42, 49, 57, 65, 72, 78, 86, 101, 111 e 118 dias após emergência (DAE. Os dados de massa de matéria seca foram ajustados ao modelo logístico e, também, utilizados para a obtenção da taxa de crescimento absoluto. Para avaliar a habilidade competitiva intra-específica e interespecífica, foram instalados cinco experimentos com o uso do método substitutivo. Compararam-se os biótipos R e S entre si e cada um destes com a cultura da soja, quando semeada no mesmo dia ou sete dias após a semeadura das plantas daninhas. As proporções de plantas entre as espécies ou biótipos utilizados foram: 5:0; 4:1; 3:2; 2:3; 1:4 e 0:5. Os biótipos de capim-colchão apresentaram acúmulo de matéria seca, crescimento absoluto e competição interespecífica semelhantes, e a redução da matéria seca da soja foi similar na presença dos biótipos R e S, o que sugere que ambos os biótipos de capim-colchão possuem a mesma adaptabilidade ecológica.The objective of this work was to compare the growth and the competitive ability of two crabgrass (Digitaria ciliaris biotypes, one resistant (R and other susceptible (S to the acetyl coenzyme A carboxylase inhibiting herbicides. Biotypes growth was determined by collecting the plant dry mass at 14, 21, 25, 28, 34, 42, 49, 57, 65, 72, 78, 86, 101, 111, and 118 days after emergence (DAE. Data of dry mass were adjusted to a logistic model, and were also used to calculate the absolute growth rate. Five experiments were installed to evaluate the intraspecific and interspecific competitive ability, using the substitutive method. The biotypes R and S were compared between

  16. Fuzzy Small Submodule and Jacobson -Radical

    OpenAIRE

    Saifur Rahman; Helen K. Saikia

    2011-01-01

    Using the notion of fuzzy small submodules of a module, we introduce the concept of fuzzy coessential extension of a fuzzy submodule of a module. We attempt to investigate various properties of fuzzy small submodules of a module. A necessary and sufficient condition for fuzzy small submodules is established. We investigate the nature of fuzzy small submodules of a module under fuzzy direct sum. Fuzzy small submodules of a module are characterized in terms of fuzzy quotient modules. This chara...

  17. Small bowel neoplasia in coeliac disease

    OpenAIRE

    Rampertab, S D; Forde, K A; Green, P. H. R.

    2003-01-01

    There is an increased risk of small bowel adenocarcinoma in patients with coeliac disease compared with the normal population. It has been suggested that adenocarcinoma of the small intestine in coeliac disease arises through an adenoma-carcinoma sequence but there has been only one reported case of a small bowel adenoma in a patient with coeliac disease. We report three additional cases of a small bowel adenoma in the setting of coeliac disease. In addition, four cases of small bowel adenoca...

  18. Small crater populations on Vesta

    CERN Document Server

    Marchi, S; O'Brien, D P; Schenk, P; Mottola, S; De Sanctis, M C; Kring, D A; Williams, D A; Raymond, C A; Russell, C T

    2013-01-01

    The NASA Dawn mission has extensively examined the surface of asteroid Vesta, the second most massive body in the main belt. The high quality of the gathered data provides us with an unique opportunity to determine the surface and internal properties of one of the most important and intriguing main belt asteroids (MBAs). In this paper, we focus on the size frequency distributions (SFDs) of sub-kilometer impact craters observed at high spatial resolution on several selected young terrains on Vesta. These small crater populations offer an excellent opportunity to determine the nature of their asteroidal precursors (namely MBAs) at sizes that are not directly observable from ground-based telescopes (i.e., below ~100 m diameter). Moreover, unlike many other MBA surfaces observed by spacecraft thus far, the young terrains examined had crater spatial densities that were far from empirical saturation. Overall, we find that the cumulative power-law index (slope) of small crater SFDs on Vesta is fairly consistent with...

  19. Railgun launch of small bodies

    Energy Technology Data Exchange (ETDEWEB)

    Drobyshevski, E.M.; Zhukov, B.G.; Sakharov, V.A. [Russian Academy of Sciences, St. Petersburg (Russian Federation). Ioffe Physico-Technical Inst.

    1995-01-01

    The small body launching using gas or plasma faces the fundamental problem caused by excess energy loss due to great wall surface/volume of the barrel ratio. That is why the efficiency of the plasma armature (PA) railgun acceleration is maximum for 8--10 mm-size bodies and drops as their size decreases. For the nuclear fusion applications, where {number_sign}1--2 mm-size pellets at 5--10 km/s velocity are desirable, one is forced to search for compromise between the body size (3--4 mm) and its velocity (3 km/s). Under these conditions, EM launchers did not demonstrate an advantage over the light-gas guns. When elaborating the {number_sign}1 mm railgun, the authors made use of the ideology of the body launching at constant acceleration close to the body strength or the electrode skin-layer explosion limits. That shortened the barrel length sufficiently. The system becomes highly compact thus permitting rapid test of new operation modes and different modifications of the design including the magnetic field augmentation. As a result of these refinements, the difficulties caused by the catastrophic supply of mass ablated from the electrodes were overcome and regimes of {number_sign}1 mm body non-sabot speed-up to 4.5 km/s were found. Potentialities of the small system created are far from being exhausted.

  20. Small gauge vitrectomy: Recent update

    Directory of Open Access Journals (Sweden)

    Sumeet Khanduja

    2013-01-01

    Full Text Available Small gauge vitrectomy, also known as minimally invasive vitreous surgery (MIVS, is a classic example of progress in biomedical engineering. Disparity in conjunctival and scleral wound location and reduction in wound diameter are its core principles. Fluidic changes include increased pressure head loss with consequent reduction in infusional flow rate and use of higher aspiration vacuum at the cutter port. Increase An increase in port open/port closed time maintains an adequate rate of vitreous removal. High Intensity Discharge (HID lamps maintain adequate illumination in spite of a decrease in the number of fiberoptic fibers. The advantages of MIVS are, a shorter surgical time, minimal conjunctival damage, and early postoperative recovery. Most complications are centered on wound stability and risk of postoperative hypotony, endophthalmitis, and port site retinal break formation. MIVS is suited in most cases, however, it can cause dehiscence of recent cataract wounds. Retraction of the infusion cannula in the suprachoroidal space may occur in eyes with scleral thinning. As a lot has been published and discussed about sutureless vitrectomy a review of this subject is necessary. A PubMed search was performed in December 2011 with terms small gauge vitrectomy, 23-gauge vitrectomy, 25-gauge vitrectomy, and 27 gauge vitrectomy, which were revised in August 2012. There were no restrictions on the date of publication but it was restricted to articles in English or other languages, if there abstracts were available in English.