WorldWideScience

Sample records for nadh dehydrogenase subunit

  1. NADH dehydrogenase subunit 1 and cytochrome c oxidase subunit I sequences compared for members of the genus Taenia (Cestoda).

    Science.gov (United States)

    Gasser, R B; Zhu, X; McManus, D P

    1999-12-01

    Nine members of the genus Taenia (Taenia taeniaeformis, Taenia hydatigena, Taenia pisiformis, Taenia ovis, Taenia multiceps, Taenia serialis, Taenia saginata, Taenia solium and the Asian Taenia) were characterised by their mitochondrial NADH dehydrogenase subunit 1 gene sequences and their genetic relationships were compared with those derived from the cytochrome c oxidase subunit 1 sequence data. The extent of inter-taxon sequence difference in NADH dehydrogenase subunit 1 (approximately 5.9-30.8%) was usually greater than in cytochrome c oxidase subunit 1 (approximately 2.5-18%). Although topology of the phenograms derived from NADH dehydrogenase subunit 1 and cytochrome c oxidase subunit 1 sequence data differed, there was concordance in that T. multiceps, T. serialis (of canids), T. saginata and the Asian Taenia (of humans) were genetically most similar, and those four members were genetically more similar to T. ovis and T. solium than they were to T. hydatigena and T. pisiformis (of canids) or T. taeniaeformis (of cats). The NADH dehydrogenase subunit 1 sequence data may prove useful in studies of the systematics and population genetic structure of the Taeniidae.

  2. Unassigned MURF1 of kinetoplastids codes for NADH dehydrogenase subunit 2

    Directory of Open Access Journals (Sweden)

    Burger Gertraud

    2008-10-01

    Full Text Available Abstract Background In a previous study, we conducted a large-scale similarity-free function prediction of mitochondrion-encoded hypothetical proteins, by which the hypothetical gene murf1 (maxicircle unidentified reading frame 1 was assigned as nad2, encoding subunit 2 of NADH dehydrogenase (Complex I of the respiratory chain. This hypothetical gene occurs in the mitochondrial genome of kinetoplastids, a group of unicellular eukaryotes including the causative agents of African sleeping sickness and leishmaniasis. In the present study, we test this assignment by using bioinformatics methods that are highly sensitive in identifying remote homologs and confront the prediction with available biological knowledge. Results Comparison of MURF1 profile Hidden Markov Model (HMM against function-known profile HMMs in Pfam, Panther and TIGR shows that MURF1 is a Complex I protein, but without specifying the exact subunit. Therefore, we constructed profile HMMs for each individual subunit, using all available sequences clustered at various identity thresholds. HMM-HMM comparison of these individual NADH subunits against MURF1 clearly identifies this hypothetical protein as NAD2. Further, we collected the relevant experimental information about kinetoplastids, which provides additional evidence in support of this prediction. Conclusion Our in silico analyses provide convincing evidence for MURF1 being a highly divergent member of NAD2.

  3. Mitochondrial genome from the facultative anaerobe and petite-positive yeast Dekkera bruxellensis contains the NADH dehydrogenase subunit genes.

    Science.gov (United States)

    Procházka, Emanuel; Poláková, Silvia; Piskur, Jure; Sulo, Pavol

    2010-08-01

    The progenitor of the Dekkera/Brettanomyces clade separated from the Saccharomyces/Kluyveromyces clade over 200 million years ago. However, within both clades, several lineages developed similar physiological traits. Both Saccharomyces cerevisiae and Dekkera bruxellensis are facultative anaerobes; in the presence of excess oxygen and sugars, they accumulate ethanol (Crabtree effect) and they both spontaneously generate respiratory-deficient mutants (petites). In order to understand the role of respiratory metabolism, the mitochondrial DNA (mtDNA) molecules of two Dekkera/Brettanomyces species were analysed. Dekkera bruxellensis mtDNA shares several properties with S. cerevisiae, such as the large genome size (76 453 bp), and the organization of the intergenic sequences consisting of spacious AT-rich regions containing a number of hairpin GC-rich cluster-like elements. In addition to a basic set of the mitochondrial genes coding for the components of cytochrome oxidase, cytochrome b, subunits of ATPase, two rRNA subunits and 25 tRNAs, D. bruxellensis also carries genes for the NADH dehydrogenase complex. Apparently, in yeast, the loss of this complex is not a precondition to develop a petite-positive, Crabtree-positive and anaerobic nature. On the other hand, mtDNA from a petite-negative Brettanomyces custersianus is much smaller (30 058 bp); it contains a similar gene set and has only short intergenic sequences.

  4. Genetic diversity of Echinococcus granulosus in southwest China determined by the mitochondrial NADH dehydrogenase subunit 2 gene.

    Science.gov (United States)

    Wang, Jiahai; Wang, Ning; Hu, Dandan; Zhong, Xiuqin; Wang, Shuxian; Gu, Xiaobin; Peng, Xuerong; Yang, Guangyou

    2014-01-01

    We evaluated genetic diversity and structure of Echinococcus granulosus by analyzing the complete mitochondrial NADH dehydrogenase subunit 2 (ND2) gene in 51 isolates of E. granulosus sensu stricto metacestodes collected at three locations in this region. We detected 19 haplotypes, which formed a distinct clade with the standard sheep strain (G1). Hence, all 51 isolates were identified as E. granulosus sensu stricto (G1-G3). Genetic relationships among haplotypes were not associated with geographical divisions, and fixation indices (Fst) among sampling localities were low. Hence, regional populations of E. granulosus in the southwest China are not differentiated, as gene flow among them remains high. This information is important for formulating unified region-wide prevention and control measures. We found large negative Fu's Fs and Tajima's D values and a unimodal mismatch distribution, indicating that the population has undergone a demographic expansion. We observed high genetic diversity among the E. granulosus s. s. isolates, indicating that the parasite population in this important bioregion is genetically robust and likely to survive and spread. The data from this study will prove valuable for future studies focusing on improving diagnosis and prevention methods and developing robust control strategies.

  5. NADH dehydrogenase subunit-2 237 Leu/Met polymorphism modifies effects of cigarette smoking on risk of elevated levels of serum liver enzyme in male Japanese health check-up examinees: a cross-sectional study

    OpenAIRE

    Kokaze, Akatsuki; Yoshida, Masao; Ishikawa, Mamoru; Matsunaga, Naomi; Karita, Kanae; Ohtsu, Tadahiro; Ochiai, Hirotaka; Shirasawa, Takako; Nanri, Hinako; Baba, Yuta; HOSHINO, Hiromi; Takashima, Yutaka

    2014-01-01

    Background NADH dehydrogenase subunit-2 237 leucine/methionine (ND2-237 Leu/Met) polymorphism reportedly influences the effects of cigarette smoking on respiratory function, risk of dyslipidemia, serum non-high-density lipoprotein cholesterol levels, hematological parameters and intraocular pressure. The objective of this study was to investigate whether ND2-237 Leu/Met polymorphism modifies the effects of cigarette smoking on serum liver enzyme levels in male Japanese health check-up examine...

  6. Identification of a new human mtDNA polymorphism (A14290G in the NADH dehydrogenase subunit 6 gene

    Directory of Open Access Journals (Sweden)

    M. Houshmand

    2006-06-01

    Full Text Available Leber's hereditary optic neuropathy (LHON is a maternally inherited form of retinal ganglion cell degeneration leading to optic atrophy in young adults. Several mutations in different genes can cause LHON (heterogeneity. The ND6 gene is one of the mitochondrial genes that encodes subunit 6 of complex I of the respiratory chain. This gene is a hot spot gene. Fourteen Persian LHON patients were analyzed with single-strand conformational polymorphism and DNA sequencing techniques. None of these patients had four primary mutations, G3460A, G11788A, T14484C, and G14459A, related to this disease. We identified twelve nucleotide substitutions, G13702C, T13879C, T14110C, C14167T, G14199T, A14233G, G14272C, A14290G, G14365C, G14368C, T14766C, and T14798C. Eleven of twelve nucleotide substitutions had already been reported as polymorphism. One of the nucleotide substitutions (A14290G has not been reported. The A14290G nucleotide substitution does not change its amino acid (glutamic acid. We looked for base conservation using DNA star software (MEGALIGN program as a criterion for pathogenic or nonpathogenic nucleotide substitution in A14290G. The results of ND6 gene alignment in humans and in other species (mouse, cow, elegans worm, and Neurospora crassa mold revealed that the 14290th base was not conserved. Fifty normal controls were also investigated for this polymorphism in the Iranian population and two had A14290G polymorphism (4%. This study provides evidence that the mtDNA A14290G allele is a new nonpathogenic polymorphism. We suggest follow-up studies regarding this polymorphism in different populations.

  7. Longevity-associated NADH Dehydrogenase Subunit-2 237 Leu/Met Polymorphism Modulates the Effects of Daily Alcohol Drinking on Yearly Changes in Serum Total and LDL Cholesterol in Japanese Men

    Directory of Open Access Journals (Sweden)

    Takashima,Yutaka

    2009-12-01

    Full Text Available Reduced nicotinamide adenine dinucleotide (NADH dehydrogenase subunit 2 237 leucine/methionine (ND2-237 Leu/Met polymorphism, is reportedly associated with longevity in the Japanese population. The ND2-237Met genotype may exert resistance to atherogenic diseases, such as myocardial infarction or cerebrovascular disorders. To investigate whether ND2-237 Leu/Met polymorphism is associated with yearly changes in serum lipid levels, we conducted a longitudinal study of 107 healthy Japanese male subjects. Analysis of covariance revealed that the interaction between the ND2-237 Leu/Met genotypes and habitual drinking was significantly associated with yearly changes in serum total cholesterol (TC and low-density lipoprotein cholesterol (LDLC levels (p0.036 and p0.006, respectively. In multiple regression analysis, daily drinking was significantly and positively associated with yearly changes in serum LDLC levels in men with ND2-237Met (p0.026. After adjusting for covariates, yearly changes in serum LDLC levels were significantly lower in non-daily drinkers with ND2-237Met than in those with ND2-237Leu (p0.047. These results suggest that ND2-237Met has a beneficial impact on yearly changes in serum LDLC in non-daily drinkers but not in daily drinkers.

  8. Fifteen novel mutations in the mitochondrial NADH dehydrogenase subunit 1, 2, 3, 4, 4L, 5 and 6 genes from Iranian patients with Leber's hereditary optic neuropathy (LHON).

    Science.gov (United States)

    Rezvani, Zahra; Didari, Elmira; Arastehkani, Ahoura; Ghodsinejad, Vadieh; Aryani, Omid; Kamalidehghan, Behnam; Houshmand, Massoud

    2013-12-01

    Leber's hereditary optic neuropathy (LHON) is an optic nerve dysfunction resulting from mutations in mitochondrial DNA (mtDNA), which is transmitted in a maternal pattern of inheritance. It is caused by three primary point mutations: G11778A, G3460A and T14484C; in the mitochondrial genome. These mutations are sufficient to induce the disease, accounting for the majority of LHON cases, and affect genes that encode for the different subunits of mitochondrial complexes I and III of the mitochondrial respiratory chain. Other mutations are secondary mutations associated with the primary mutations. The purpose of this study was to determine MT-ND variations in Iranian patients with LHON. In order to determine the prevalence and distribution of mitochondrial mutations in the LHON patients, their DNA was studied using PCR and DNA sequencing analysis. Sequencing of MT-ND genes from 35 LHON patients revealed a total of 44 nucleotide variations, in which fifteen novel variations-A14020G, A13663G, C10399T, C4932A, C3893G, C10557A, C12012A, C13934T, G4596A, T12851A, T4539A, T4941A, T13255A, T14353C and del A 4513-were observed in 27 LHON patients. However, eight patients showed no variation in the ND genes. These mutations contribute to the current database of mtDNA polymorphisms in LHON patients and may facilitate the definition of disease-related mutations in human mtDNA. This research may help to understand the disease mechanism and open up new diagnostic opportunities for LHON.

  9. Reduced mitochondrial Ca(2+) transients stimulate autophagy in human fibroblasts carrying the 13514A>G mutation of the ND5 subunit of NADH dehydrogenase.

    Science.gov (United States)

    Granatiero, V; Giorgio, V; Calì, T; Patron, M; Brini, M; Bernardi, P; Tiranti, V; Zeviani, M; Pallafacchina, G; De Stefani, D; Rizzuto, R

    2016-02-01

    Mitochondrial disorders are a group of pathologies characterized by impairment of mitochondrial function mainly due to defects of the respiratory chain and consequent organellar energetics. This affects organs and tissues that require an efficient energy supply, such as brain and skeletal muscle. They are caused by mutations in both nuclear- and mitochondrial DNA (mtDNA)-encoded genes and their clinical manifestations show a great heterogeneity in terms of age of onset and severity, suggesting that patient-specific features are key determinants of the pathogenic process. In order to correlate the genetic defect to the clinical phenotype, we used a cell culture model consisting of fibroblasts derived from patients with different mutations in the mtDNA-encoded ND5 complex I subunit and with different severities of the illness. Interestingly, we found that cells from patients with the 13514A>G mutation, who manifested a relatively late onset and slower progression of the disease, display an increased autophagic flux when compared with fibroblasts from other patients or healthy donors. We characterized their mitochondrial phenotype by investigating organelle turnover, morphology, membrane potential and Ca(2+) homeostasis, demonstrating that mitochondrial quality control through mitophagy is upregulated in 13514A>G cells. This is due to a specific downregulation of mitochondrial Ca(2+) uptake that causes the stimulation of the autophagic machinery through the AMPK signaling axis. Genetic and pharmacological manipulation of mitochondrial Ca(2+) homeostasis can revert this phenotype, but concurrently decreases cell viability. This indicates that the higher mitochondrial turnover in complex I deficient cells with this specific mutation is a pro-survival compensatory mechanism that could contribute to the mild clinical phenotype of this patient.

  10. The Kinetic Reaction Mechanism of the Vibrio cholerae Sodium-dependent NADH Dehydrogenase*♦

    Science.gov (United States)

    Tuz, Karina; Mezic, Katherine G.; Xu, Tianhao; Barquera, Blanca; Juárez, Oscar

    2015-01-01

    The sodium-dependent NADH dehydrogenase (Na+-NQR) is the main ion transporter in Vibrio cholerae. Its activity is linked to the operation of the respiratory chain and is essential for the development of the pathogenic phenotype. Previous studies have described different aspects of the enzyme, including the electron transfer pathways, sodium pumping structures, cofactor and subunit composition, among others. However, the mechanism of the enzyme remains to be completely elucidated. In this work, we have studied the kinetic mechanism of Na+-NQR with the use of steady state kinetics and stopped flow analysis. Na+-NQR follows a hexa-uni ping-pong mechanism, in which NADH acts as the first substrate, reacts with the enzyme, and the oxidized NAD leaves the catalytic site. In this conformation, the enzyme is able to capture two sodium ions and transport them to the external side of the membrane. In the last step, ubiquinone is bound and reduced, and ubiquinol is released. Our data also demonstrate that the catalytic cycle involves two redox states, the three- and five-electron reduced forms. A model that gathers all available information is proposed to explain the kinetic mechanism of Na+-NQR. This model provides a background to understand the current structural and functional information. PMID:26004776

  11. NADH dehydrogenase-like behavior of nitrogen-doped graphene and its application in NAD(+)-dependent dehydrogenase biosensing.

    Science.gov (United States)

    Gai, Pan-Pan; Zhao, Cui-E; Wang, Ying; Abdel-Halim, E S; Zhang, Jian-Rong; Zhu, Jun-Jie

    2014-12-15

    A novel electrochemical biosensing platform for nicotinamide adenine dinucleotide (NAD(+))-dependent dehydrogenase catalysis was designed using the nitrogen-doped graphene (NG), which had properties similar to NADH dehydrogenase (CoI). NG mimicked flavin mononucleotide (FMN) in CoI and efficiently catalyzed NADH oxidation. NG also acted as an electron transport "bridge" from NADH to the electrode due to its excellent conductivity. In comparison with a bare gold electrode, an 800 mV decrease in the overpotential for NADH oxidation and CoI-like behavior were observed at NG-modified electrode, which is the largest decrease in overpotential for NADH oxidation reported to date. The catalytic rate constant (k) for the CoI-like behavior of NG was estimated to be 2.3×10(5) M(-1) s(-1), which is much higher than that of other previously reported FMN analogs. The Michaelis-Menten constant (Km) of NG was 26 μM, which is comparable to the Km of CoI (10 μM). Electrodes modified with NG and NG/gold nanoparticals/formate dehydrogenase (NG/AuNPs/FDH) showed excellent analytical performance for the detection of NADH and formate. This electrode fabrication strategy could be used to create a universal biosensing platform for developing NAD(+)-dependent dehydrogenase biosensors and biofuel cells. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. In situ Regeneration of NADH via Lipoamide Dehydrogenase-catalyzed Electron Transfer Reaction Evidenced by Spectroelectrochemistry

    Energy Technology Data Exchange (ETDEWEB)

    Tam, Tsz Kin; Chen, Baowei; Lei, Chenghong; Liu, Jun

    2012-08-01

    NAD/NADH is a coenzyme found in all living cells, carrying electrons from one reaction to another. We report on characterizations of in situ regeneration of NADH via lipoamide dehydrogenase (LD)-catalyzed electron transfer reaction to regenerate NADH using UV-vis spectroelectrochemistry. The Michaelis-Menten constant (Km) and maximum velocity (Vmax) of NADH regeneration were measured as 0.80 {+-} 0.15 mM and 1.91 {+-} 0.09 {micro}M s-1 in a 1-mm thin-layer spectroelectrochemical cell using gold gauze as the working electrode at the applied potential -0.75 V (vs. Ag/AgCl). The electrocatalytic reduction of the NAD system was further coupled with the enzymatic conversion of pyruvate to lactate by lactate dehydrogenase to examine the coenzymatic activity of the regenerated NADH. Although the reproducible electrocatalytic reduction of NAD into NADH is known to be difficult compared to the electrocatalytic oxidation of NADH, our spectroelectrochemical results indicate that the in situ regeneration of NADH via LD-catalyzed electron transfer reaction is fast and sustainable and can be potentially applied to many NAD/NADH-dependent enzyme systems.

  13. Initial Evidence for Adaptive Selection on the NADH Subunit Two of Freshwater Dolphins by Analyses of Mitochondrial Genomes

    Science.gov (United States)

    Caballero, Susana; Duchêne, Sebastian; Garavito, Manuel F.; Slikas, Beth; Baker, C. Scott

    2015-01-01

    A small number of cetaceans have adapted to an entirely freshwater environment, having colonized rivers in Asia and South America from an ancestral origin in the marine environment. This includes the ‘river dolphins’, early divergence from the odontocete lineage, and two species of true dolphins (Family Delphinidae). Successful adaptation to the freshwater environment may have required increased demands in energy involved in processes such as the mitochondrial osmotic balance. For this reason, riverine odontocetes provide a compelling natural experiment in adaptation of mammals from marine to freshwater habitats. Here we present initial evidence of positive selection in the NADH dehydrogenase subunit 2 of riverine odontocetes by analyses of full mitochondrial genomes, using tests of selection and protein structure modeling. The codon model with highest statistical support corresponds to three discrete categories for amino acid sites, those under positive, neutral, and purifying selection. With this model we found positive selection at site 297 of the NADH dehydrogenase subunit 2 (dN/dS>1.0,) leading to a substitution of an Ala or Val from the ancestral state of Thr. A phylogenetic reconstruction of 27 cetacean mitogenomes showed that an Ala substitution has evolved at least four times in cetaceans, once or more in the three ‘river dolphins’ (Families Pontoporidae, Lipotidae and Inidae), once in the riverine Sotalia fluviatilis (but not in its marine sister taxa), once in the riverine Orcaella brevirostris from the Mekong River (but not in its marine sister taxa) and once in two other related marine dolphins. We located the position of this amino acid substitution in an alpha-helix channel in the trans-membrane domain in both the E. coli structure and Sotalia fluviatilis model. In E. coli this position is located in a helix implicated in a proton translocation channel of respiratory complex 1 and may have a similar role in the NADH dehydrogenases of

  14. Initial Evidence for Adaptive Selection on the NADH Subunit Two of Freshwater Dolphins by Analyses of Mitochondrial Genomes.

    Directory of Open Access Journals (Sweden)

    Susana Caballero

    Full Text Available A small number of cetaceans have adapted to an entirely freshwater environment, having colonized rivers in Asia and South America from an ancestral origin in the marine environment. This includes the 'river dolphins', early divergence from the odontocete lineage, and two species of true dolphins (Family Delphinidae. Successful adaptation to the freshwater environment may have required increased demands in energy involved in processes such as the mitochondrial osmotic balance. For this reason, riverine odontocetes provide a compelling natural experiment in adaptation of mammals from marine to freshwater habitats. Here we present initial evidence of positive selection in the NADH dehydrogenase subunit 2 of riverine odontocetes by analyses of full mitochondrial genomes, using tests of selection and protein structure modeling. The codon model with highest statistical support corresponds to three discrete categories for amino acid sites, those under positive, neutral, and purifying selection. With this model we found positive selection at site 297 of the NADH dehydrogenase subunit 2 (dN/dS>1.0, leading to a substitution of an Ala or Val from the ancestral state of Thr. A phylogenetic reconstruction of 27 cetacean mitogenomes showed that an Ala substitution has evolved at least four times in cetaceans, once or more in the three 'river dolphins' (Families Pontoporidae, Lipotidae and Inidae, once in the riverine Sotalia fluviatilis (but not in its marine sister taxa, once in the riverine Orcaella brevirostris from the Mekong River (but not in its marine sister taxa and once in two other related marine dolphins. We located the position of this amino acid substitution in an alpha-helix channel in the trans-membrane domain in both the E. coli structure and Sotalia fluviatilis model. In E. coli this position is located in a helix implicated in a proton translocation channel of respiratory complex 1 and may have a similar role in the NADH dehydrogenases of

  15. Chloroplast NDH: A different enzyme with a structure similar to that of respiratory NADH dehydrogenase.

    Science.gov (United States)

    Shikanai, Toshiharu

    2016-07-01

    Eleven genes encoding chloroplast NADH dehydrogenase-like (NDH) complex have been discovered in plastid genomes on the basis of their homology with genes encoding respiratory complex I. Despite this structural similarity, chloroplast NDH and its evolutionary origin NDH-1 in cyanobacteria accept electrons from ferredoxin (Fd), indicating that chloroplast NDH is an Fd-dependent plastoquinone (PQ) reductase rather than an NAD(P)H dehydrogenase. In Arabidopsis thaliana, chloroplast NDH interacts with photosystem I (PSI); this interaction is needed to stabilize NDH, especially under high light. On the basis of these distinct characters of chloroplast and cyanobacterial NDH, it can be distinguished as a photosynthetic NDH from respiratory complex I. In fact, chloroplast NDH forms part of the machinery of photosynthesis by mediating the minor pathway of PSI cyclic electron transport. Along with the antimycin A-sensitive main pathway of PSI cyclic electron transport, chloroplast NDH compensates the ATP/NADPH production ratio in the light reactions of photosynthesis. In this review, I revisit the original concept of chloroplast NDH on the basis of its similarity to respiratory complex I and thus introduce current progress in the field to researchers focusing on respiratory complex I. I summarize recent progress on the basis of structure and function. Finally, I introduce the results of our examination of the process of assembly of chloroplast NDH. Although the process requires many plant-specific non-subunit factors, the core processes of assembly are conserved between chloroplast NDH and respiratory complex I. This article is part of a Special Issue entitled Respiratory complex I, edited by Volker Zickermann and Ulrich Brandt. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. 湖南省泡状带绦虫线粒体nad1基因的序列测定及分析%Sequence Measurement and Analysis of Mitochondrial NADH Dehydrogenase Subunit 1 of Taenia Hydatigena Collected from Hunan Province

    Institute of Scientific and Technical Information of China (English)

    伍慧兰

    2011-01-01

    To analyze the mitochondrial NADH dehydrogenase subunit 1 (nadI) gene of Taenia hydatigena collected from Hunan province, one should follow the three steps. Firstly, the partial nadl (pnadl) is amplified from each Taen/a hyclatigena sample. Then pnadl sequences are aligned by using the ClustalX 1.81. Lastly, sequence homology analyfis is conducted by using the Megalign program of the software DNAStar version 5.0. The result shows that the length of pnadl is 391bp. This result has provided a foundation for further studies of molecular identification and molecular genetics of Taenia hydatigena.%以从我国湖南长沙和湘西犬小肠中采集的2条泡状带绦虫作为研究对象,用引物JB11及JB12扩增泡状带绦虫的pnad1片段,应用ClustalX1.81程序对序列进行比对,同时利用DNAscar5.0中的Megalign程序进行同源性分析。结果显示来自湖南长沙和湘西的2条泡状带绦虫的pnad1序列均为391bp。研究结果为泡状带绦虫进一步的分类、鉴定和遗传变异研究奠定了基础。

  17. New complexes containing the internal alternative NADH dehydrogenase (Ndi1) in mitochondria of Saccharomyces cerevisiae.

    Science.gov (United States)

    Matus-Ortega, M G; Cárdenas-Monroy, C A; Flores-Herrera, O; Mendoza-Hernández, G; Miranda, M; González-Pedrajo, B; Vázquez-Meza, H; Pardo, J P

    2015-10-01

    Mitochondria of Saccharomyces cerevisiae lack the respiratory complex I, but contain three rotenone-insensitive NADH dehydrogenases distributed on both the external (Nde1 and Nde2) and internal (Ndi1) surfaces of the inner mitochondrial membrane. These enzymes catalyse the transfer of electrons from NADH to ubiquinone without the translocation of protons across the membrane. Due to the high resolution of the Blue Native PAGE (BN-PAGE) technique combined with digitonin solubilization, several bands with NADH dehydrogenase activity were observed on the gel. The use of specific S. cerevisiae single and double mutants of the external alternative elements (ΔNDE1, ΔNDE2, ΔNDE1/ΔNDE2) showed that the high and low molecular weight complexes contained the Ndi1. Some of the Ndi1 associations took place with complexes III and IV, suggesting the formation of respirasome-like structures. Complex II interacted with other proteins to form a high molecular weight supercomplex with a molecular mass around 600 kDa. We also found that the majority of the Ndi1 was in a dimeric form, which is in agreement with the recently reported three-dimensional structure of the protein.

  18. Optical isopropanol biosensor using NADH-dependent secondary alcohol dehydrogenase (S-ADH).

    Science.gov (United States)

    Chien, Po-Jen; Ye, Ming; Suzuki, Takuma; Toma, Koji; Arakawa, Takahiro; Iwasaki, Yasuhiko; Mitsubayashi, Kohji

    2016-10-01

    Isopropanol (IPA) is an important solvent used in industrial activity often found in hospitals as antiseptic alcohol rub. Also, IPA may have the potential to be a biomarker of diabetic ketoacidosis. In this study, an optical biosensor using NADH-dependent secondary alcohol dehydrogenase (S-ADH) for IPA measurement was constructed and evaluated. An ultraviolet light emitting diode (UV-LED, λ=340nm) was employed as the excitation light to excite nicotinamide adenine dinucleotide (NADH). A photomultiplier tube (PMT) was connected to a two-way branch optical fiber for measuring the fluorescence emitted from the NADH. S-ADH was immobilized on the membrane to catalyze IPA to acetone and reduce NAD(+) to be NADH. This IPA biosensor shows highly sensitivity and selectivity, the calibration range is from 500 nmol L(-1) to 1mmolL(-1). The optimization of buffer pH, temperature, and the enzyme-immobilized method were also evaluated. The detection of IPA in nail related cosmetic using our IPA biosensor was also carried out. The results showed that large amounts of IPA were used in these kinds of cosmetics. This IPA biosensor comes with the advantages of rapid reaction, good reproducibility, and wide dynamic range, and is also expected to use for clinical IPA detections in serum or other medical and health related applications.

  19. Aqueous soluble tetrazolium/formazan MTS as an indicator of NADH- and NADPH-dependent dehydrogenase activity.

    Science.gov (United States)

    Dunigan, D D; Waters, S B; Owen, T C

    1995-10-01

    Recently a new tetrazolium was described for the use of monitoring cell viability in culture. This tetrazolium, commonly referred to as MTS [3-(4,5-dimethylthiazol-2-yl)- 5-(3-carboxymethonyphenol)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt], has the unusual property that it can be reduced to a water-soluble formazan. beta-Nicotinamide adenine dinucleotide/reduced (NADH) and beta-nicotinamide adenine dinucleotide phosphate/reduced (NADPH) are examples of physiologically important reducing agents. In cell-free studies, MTS was reduce to the soluble formazan in the presence of NADH and NADPH, and reaction were compared to those with dithiothreitol (DTT) or 2-mercaptoethanol (2-ME). The efficiency of these reactions was enhanced 1000-fold by the presence of phenazine methosulfate. Selectivity in the electron transfer from NADPH was slightly greater than NADH, and NADPH or NADH was much greater than the thiols DTT or 2-ME. Generation of either NADH or NADPH in solution by malate dehydrogenase or isocitrate dehydrogenase, respectively, was monitored by the MTS reduction reaction. The rate of formazan formation was comparable to the formation of NADH or NADPH. This system represents a useful tool for evaluating reaction kinetics in solutions of NAD- or NADP-dependent dehydrogenase enzymes, and these reactions can be performed in typical biological buffers containing reducing agents without significant interference to the MTS/formazan system.

  20. Purification of two putative type II NADH dehydrogenases with different substrate specificities from alkaliphilic Bacillus pseudofirmus OF4.

    Science.gov (United States)

    Liu, Jun; Krulwich, Terry A; Hicks, David B

    2008-05-01

    A putative Type II NADH dehydrogenase from Halobacillus dabanensis was recently reported to have Na+/H+ antiport activity (and called Nap), raising the possibility of direct coupling of respiration to antiport-dependent pH homeostasis. This study characterized a homologous type II NADH dehydrogenase of genetically tractable alkaliphilic Bacillus pseudofirmus OF4, in which evidence supports antiport-based pH homeostasis that is mediated entirely by secondary antiport. Two candidate type II NADH dehydrogenase genes with canonical GXGXXG motifs were identified in a draft genome sequence of B. pseudofirmus OF4. The gene product designated NDH-2A exhibited homology to enzymes from Bacillus subtilis and Escherichia coli whereas NDH-2B exhibited homology to the H. dabanensis Nap protein and its alkaliphilic Bacillus halodurans C-125 homologue. The ndh-2A, but not the ndh-2B, gene complemented the growth defect of an NADH dehydrogenase-deficient E. coli mutant. Neither gene conferred Na+-resistance on an antiporter-deficient E. coli strain, nor did they confer Na+/H+ antiport activity in vesicle assays. The purified hexa-histidine-tagged gene products were approximately 50 kDa, contained noncovalently bound FAD and oxidized NADH. They were predominantly cytoplasmic in E. coli, consonant with the absence of antiport activity. The catalytic properties of NDH-2A were more consistent with a major respiratory role than those of NDH-2B.

  1. The kinetics behavior of the reduction of formaldehyde catalyzed by Alcohol Dehydrogenase (ADH) and partial uncompetitive substrate inhibition by NADH.

    Science.gov (United States)

    Wen, Nuan; Liu, Wenfang; Hou, Yanhui; Zhao, Zhiping

    2013-05-01

    Alcohol dehydrogenase (ADH) catalyzes the final step in the biosynthesis of methanol from CO2. Here, we report the steady-state kinetics for ADH, using a homogeneous enzyme preparation with formaldehyde as the substrate and nicotinamide adenine dinucleotide (NADH) as the cofactor. When changing NADH concentrations with the fixed concentrations of HCHO (more or less than NADH), kinetic studies revealed a particular zigzag phenomenon for the first time. Increasing formaldehyde concentration can weaken substrate inhibition and improve catalytic efficiency. The kinetic mechanism of ADH was analyzed using the secondary fitting method. The double reciprocal plots (1/v∼1/[HCHO] and 1/[NADH]) strongly demonstrated that the substrate inhibition by NADH was uncompetitive versus formaldehyde and partial. In the direction of formaldehyde reduction, ADH has an ordered kinetic mechanism with formaldehyde adding to enzyme first and product methanol released last. The second reactant NADH can combine with the enzyme-methanol complex and then methanol dissociates from it at a slower rate than from enzyme-methanol. The reaction velocity depends on the relative rates of the alternative pathways. The addition of NADH also accelerates the releasing of methanol. As a result, substrate inhibition and activation occurred intermittently, and the zigzag double reciprocal plot (1/v∼1/[NADH]) was obtained.

  2. Highly stable and reusable immobilized formate dehydrogenases: Promising biocatalysts for in situ regeneration of NADH

    Directory of Open Access Journals (Sweden)

    Barış Binay

    2016-02-01

    Full Text Available This study aimed to prepare robust immobilized formate dehydrogenase (FDH preparations which can be used as effective biocatalysts along with functional oxidoreductases, in which in situ regeneration of NADH is required. For this purpose, Candida methylica FDH was covalently immobilized onto Immobead 150 support (FDHI150, Immobead 150 support modified with ethylenediamine and then activated with glutaraldehyde (FDHIGLU, and Immobead 150 support functionalized with aldehyde groups (FDHIALD. The highest immobilization yield and activity yield were obtained as 90% and 132%, respectively when Immobead 150 functionalized with aldehyde groups was used as support. The half-life times (t1/2 of free FDH, FDHI150, FDHIGLU and FDHIALD were calculated as 10.6, 28.9, 22.4 and 38.5 h, respectively at 35 °C. FDHI150, FDHIGLU and FDHIALD retained 69, 38 and 51% of their initial activities, respectively after 10 reuses. The results show that the FDHI150, FDHIGLU and FDHIALD offer feasible potentials for in situ regeneration of NADH.

  3. FAD binding properties of a cytosolic version of Escherichia coli NADH dehydrogenase-2.

    Science.gov (United States)

    Villegas, Josefina M; Valle, Lorena; Morán Vieyra, Faustino E; Rintoul, María R; Borsarelli, Claudio D; Rapisarda, Viviana A

    2014-03-01

    Respiratory NADH dehydrogenase-2 (NDH-2) of Escherichia coli is a peripheral membrane-bound flavoprotein. By eliminating its C-terminal region, a water soluble truncated version was obtained in our laboratory. Overall conformation of the mutant version resembles the wild-type protein. Considering these data and the fact that the mutant was obtained as an apo-protein, the truncated version is an ideal model to study the interaction between the enzyme and its cofactor. Here, the FAD binding properties of this version were characterized using far-UV circular dichroism (CD), differential scanning calorimetry (DSC), limited proteolysis, and steady-state and dynamic fluorescence spectroscopy. CD spectra, thermal unfolding and DSC profiles did not reveal any major difference in secondary structure between apo- and holo-protein. In addition, digestion site accessibility and tertiary conformation were similar for both proteins, as seen by comparable chymotryptic cleavage patterns. FAD binding to the apo-protein produced a parallel increment of both FAD fluorescence quantum yield and steady-state emission anisotropy. On the other hand, addition of FAD quenched the intrinsic fluorescence emission of the truncated protein, indicating that the flavin cofactor should be closely located to the protein Trp residues. Analysis of the steady-state and dynamic fluorescence data confirms the formation of the holo-protein with a 1:1 binding stoichiometry and an association constant KA=7.0(±0.8)×10(4)M(-1). Taken together, the FAD-protein interaction is energetically favorable and the addition of FAD is not necessary to induce the enzyme folded state. For the first time, a detailed characterization of the flavin:protein interaction was performed among alternative NADH dehydrogenases.

  4. Properties and subunit structure of pig heart pyruvate dehydrogenase.

    Science.gov (United States)

    Hamada, M; Hiraoka, T; Koike, K; Ogasahara, K; Kanzaki, T

    1976-06-01

    Pyruvate dehydrogenase [EC 1.2.4.1] was separated from the pyruvate dehydrogenase complex and its molecular weight was estimated to be about 150,000 by sedimentation equilibrium methods. The enzyme was dissociated into two subunits (alpha and beta), with estimated molecular weights of 41,000 (alpha) and 36,000 (beta), respectively, by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The subunits were separated by phosphocellulose column chromatography and their chemical properties were examined. The subunit structure of the pyruvate dehydrogenase was assigned as alpha2beta2. The content of right-handed alpha-helix in the enzyme molecule was estimated to be about 29 and 28% by optical rotatory dispersion and by circular dichroism, respectively. The enzyme contained no thiamine-PP, and its dehydrogenase activity was completely dependent on added thiamine-PP and partially dependent on added Mg2+ and Ca2+. The Km value of pyruvate dehydrogenase for thiamine diphosphate was estimated to be 6.5 X 10(-5) M in the presence of Mg2+ or Ca2+. The enzyme showed highly specific activity for thiamine-PP dependent oxidation of both pyruvate and alpha-ketobutyrate, but it also showed some activity with alpha-ketovalerate, alpha-ketoisocaproate, and alpha-ketoisovalerate. The pyruvate dehydrogenase activity was strongly inhibited by bivalent heavy metal ions and by sulfhydryl inhibitors; and the enzyme molecule contained 27 moles of 5,5'-dithiobis(2-nitrobenzoic acid)-reactive sulfhydryl groups and a total of 36 moles of sulfhydryl groups. The inhibitory effect of p-chloromercuribenzoate was prevented by preincubating the enzyme with thiamine-PP plus pyruvate. The structure of pyruvate dehydrogenase necessary for formation of the complex is also reported.

  5. The Na+-translocating NADH:quinone oxidoreductase (Na+-NQR) from Vibrio cholerae enhances insertion of FeS in overproduced NqrF subunit.

    Science.gov (United States)

    Tao, Minli; Fritz, Günter; Steuber, Julia

    2008-01-01

    The Na+-translocating NADH:quinone oxidoreductase (Na+-NQR) from Vibrio cholerae is a membrane-bound, respiratory Na+ pump. Its NqrF subunit contains one FAD and a [2Fe-2S] cluster and catalyzes the initial oxidation of NADH. A soluble variant of NqrF lacking its hydrophobic, N-terminal helix (NqrF') was produced in V. cholerae wild type and nqr deletion strain. Under identical conditions of growth and induction, the yield of NqrF' increased by 30% in the presence of the Na+-NQR. FAD-containing NqrF' species with or without the FeS cluster were observed, indicating that assembly of the FeS center, but not insertion of the flavin cofactor, was limited during overproduction in V. cholerae. A comparison of these distinct NqrF' species with regard to specific NADH dehydrogenase activity, pH dependence of activity and thermal inactivation showed that NqrF' lacking the [2Fe-2S] cluster was less stable, partially unfolded, and therefore prone to proteolytic degradation in V. cholerae. We conclude that the overall yield of NqrF' critically depends on the amount of fully assembled, FeS-containing NqrF' in the V. cholerae host cells. The Na+-NQR is proposed to increase the stability of NqrF' by stimulating the maturation of FeS centers.

  6. Metabolic impact of an NADH-producing glucose-6-phosphate dehydrogenase in Escherichia coli

    DEFF Research Database (Denmark)

    Olavarria, K.; De Ingeniis, J.; Zielinski, D. C.

    2014-01-01

    PDH(R46E,Q47E). Through homologous recombination, the zwf loci (encoding G6PDH) in the chromosomes of WT and Δpgi E. coli strains were replaced by DNA encoding LmG6PDH(R46E,Q47E). Contrary to some predictions performed with flux balance analysis, the replacements caused a substantial effect......, we studied the metabolic response of this bacterium to the replacement of its glucose-6-phosphate dehydrogenase (G6PDH) by an NADH-producing variant. The homologous enzyme from Leuconostoc mesenteroides was studied by molecular dynamics and site-directed mutagenesis to obtain the NAD-preferring LmG6...... on the growth rates, increasing 59 % in the Δpgi strain, while falling 44 % in the WT. Quantitative PCR (qPCR) analysis of the zwf locus showed that the expression level of the mutant enzyme was similar to the native enzyme and the expression of genes encoding key enzymes of the central pathways also showed...

  7. Life without complex I: proteome analyses of an Arabidopsis mutant lacking the mitochondrial NADH dehydrogenase complex.

    Science.gov (United States)

    Fromm, Steffanie; Senkler, Jennifer; Eubel, Holger; Peterhänsel, Christoph; Braun, Hans-Peter

    2016-05-01

    The mitochondrial NADH dehydrogenase complex (complex I) is of particular importance for the respiratory chain in mitochondria. It is the major electron entry site for the mitochondrial electron transport chain (mETC) and therefore of great significance for mitochondrial ATP generation. We recently described an Arabidopsis thaliana double-mutant lacking the genes encoding the carbonic anhydrases CA1 and CA2, which both form part of a plant-specific 'carbonic anhydrase domain' of mitochondrial complex I. The mutant lacks complex I completely. Here we report extended analyses for systematically characterizing the proteome of the ca1ca2 mutant. Using various proteomic tools, we show that lack of complex I causes reorganization of the cellular respiration system. Reduced electron entry into the respiratory chain at the first segment of the mETC leads to induction of complexes II and IV as well as alternative oxidase. Increased electron entry at later segments of the mETC requires an increase in oxidation of organic substrates. This is reflected by higher abundance of proteins involved in glycolysis, the tricarboxylic acid cycle and branched-chain amino acid catabolism. Proteins involved in the light reaction of photosynthesis, the Calvin cycle, tetrapyrrole biosynthesis, and photorespiration are clearly reduced, contributing to the significant delay in growth and development of the double-mutant. Finally, enzymes involved in defense against reactive oxygen species and stress symptoms are much induced. These together with previously reported insights into the function of plant complex I, which were obtained by analysing other complex I mutants, are integrated in order to comprehensively describe 'life without complex I'.

  8. Identity of the subunits and the stoicheiometry of prosthetic groups in trimethylamine dehydrogenase and dimethylamine dehydrogenase.

    Science.gov (United States)

    Kasprzak, A A; Papas, E J; Steenkamp, D J

    1983-01-01

    Trimethylamine dehydrogenases from bacterium W3A1 and Hyphomicrobium X and the dimethylamine dehydrogenase from Hyphomicrobium X were found to contain only one kind of subunit. The millimolar absorption coefficient of a single [4Fe-4S] cluster in trimethylamine dehydrogenase from bacterium W3A1 was estimated to be 14.8 mM-1 . cm-1 at 443 nm. From this value a 1:1 stoicheiometry of the prosthetic groups, 6-S-cysteinyl-FMN and the [4Fe-4S] cluster, was established. Millimolar absorption coefficients of the three enzymes were in the range 49.4-58.7 mM-1 . cm-1 at approx. 440 nm. This range of values is consistent with the presence of two [4Fe-4S] clusters and two flavin residues, for which the millimolar absorption coefficient had earlier been found to be 12.3 mM-1 . cm-1 at 437 nm. The N-terminal amino acid was alanine in each of the three enzymes. Sequence analysis of the first 15 residues from the N-terminus of dimethylamine dehydrogenase indicated a single unique sequence. Two identical subunits, each containing covalently bound 6-S-cysteinyl-FMN and a [4Fe-4S] cluster, in each of the enzymes are therefore indicated. Images Fig. 1. PMID:6882357

  9. Annotated compound data for modulators of detergent-solubilised or lipid-reconstituted respiratory type II NADH dehydrogenase activity obtained by compound library screening

    OpenAIRE

    Dunn, Elyse A.; Cook, Gregory M.; Adam Heikal

    2015-01-01

    The energy-generating membrane protein NADH dehydrogenase (NDH-2), a proposed antibacterial drug target (see “Inhibitors of type II NADH:menaquinone oxidoreductase represent a class of antitubercular drugs” Weinstein et al. 2005 [1]), was screened for modulators of activity in either detergent-solublised or lipid reconstituted (proteolipsome) form. Here we present an annotated list of compounds identified in a small-scale screen against NDH-2. The dataset contains information regarding the li...

  10. Crystal Structure of Human Dihydrolipoamide Dehydrogenase: NAD[superscript +]/NADH Binding and the Structural Basis of Disease-causing Mutations

    Energy Technology Data Exchange (ETDEWEB)

    Brautigam, Chad A.; Chuang, Jacinta L.; Tomchick, Diana R.; Machius, Mischa; Chuang, David T. (U. of Texas-SMED)

    2010-07-13

    Human dihydrolipoamide dehydrogenase (hE3) is an enzymatic component common to the mitochondrial {alpha}-ketoacid dehydrogenase and glycine decarboxylase complexes. Mutations to this homodimeric flavoprotein cause the often-fatal human disease known as E3 deficiency. To catalyze the oxidation of dihydrolipoamide, hE3 uses two molecules: noncovalently bound FAD and a transiently bound substrate, NAD{sup +}. To address the catalytic mechanism of hE3 and the structural basis for E3 deficiency, the crystal structures of hE3 in the presence of NAD{sup +} or NADH have been determined at resolutions of 2.5 {angstrom} and 2.1 {angstrom}, respectively. Although the overall fold of the enzyme is similar to that of yeast E3, these two structures differ at two loops that protrude from the proteins and at their FAD-binding sites. The structure of oxidized hE3 with NAD{sup +} bound demonstrates that the nicotinamide moiety is not proximal to the FAD. When NADH is present, however, the nicotinamide base stacks directly on the isoalloxazine ring system of the FAD. This is the first time that this mechanistically requisite conformation of NAD{sup +} or NADH has been observed in E3 from any species. Because E3 structures were previously available only from unicellular organisms, speculations regarding the molecular mechanisms of E3 deficiency were based on homology models. The current hE3 structures show directly that the disease-causing mutations occur at three locations in the human enzyme: the dimer interface, the active site, and the FAD and NAD{sup +}-binding sites. The mechanisms by which these mutations impede the function of hE3 are discussed.

  11. Roles of subunit NuoL in the proton pumping coupling mechanism of NADH:ubiquinone oxidoreductase (complex I) from Escherichia coli.

    Science.gov (United States)

    Narayanan, Madhavan; Sakyiama, Joseph A; Elguindy, Mahmoud M; Nakamaru-Ogiso, Eiko

    2016-10-01

    Respiratory complex I has an L-shaped structure formed by the hydrophilic arm responsible for electron transfer and the membrane arm that contains protons pumping machinery. Here, to gain mechanistic insights into the role of subunit NuoL, we investigated the effects of Mg(2+), Zn(2+) and the Na(+)/H(+) antiporter inhibitor 5-(N-ethyl-N-isopropyl)-amiloride (EIPA) on proton pumping activities of various isolated NuoL mutant complex I after reconstitution into Escherichia coli double knockout (DKO) membrane vesicles lacking complex I and the NADH dehydrogenase type 2. We found that Mg(2+) was critical for proton pumping activity of complex I. At 2 µM Zn(2+), proton pumping of the wild-type was selectively inhibited without affecting electron transfer; no inhibition in proton pumping of D178N and D400A was observed, suggesting the involvement of these residues in Zn(2+) binding. Fifteen micromolar of EIPA caused up to ∼40% decrease in the proton pumping activity of the wild-type, D303A and D400A/E, whereas no significant change was detected in D178N, indicating its possible involvement in the EIPA binding. Furthermore, when menaquinone-rich DKO membranes were used, the proton pumping efficiency in the wild-type was decreased significantly (∼50%) compared with NuoL mutants strongly suggesting that NuoL is involved in the high efficiency pumping mechanism in complex I.

  12. Characterization of the NADH-linked acetylacetoin reductase/2,3-butanediol dehydrogenase gene from Bacillus cereus YUF-4.

    Science.gov (United States)

    Hosaka, T; Ui, S; Ohtsuki, T; Mimura, A; Ohkuma, M; Kudo, T

    2001-01-01

    A 1.4-kbp DNA fragment, including the NADH-linked acetylacetoin reductase/2,3-butanediol dehydrogenase (AACRII/BDH) gene from the chromosomal DNA of Bacillus cereus YUF-4, was cloned in Escherichia coli DH5alpha after its insertion into pUC119, and the resulting plasmid was named pAACRII119. The AACRII/BDH gene had an open reading frame consisting of 1047 bp encoding 349 amino acids. The enzyme exhibited not only AACR activity, but also BDH activity. However, the gene was not located in a 2,3-butanediol (BD) operon, as is the case in the BDH gene of Klebsiella pneumoniae and that of K. terrigena. In addition, there was no BD-cycle-related enzyme gene in the region surrounding the AACRII/BDH gene. The AACR and BDH activities in E. coli DH5alpha/pAACRII119 were 200-fold higher than those in the original B. cereus YUF-4. The characteristics of the AACRII/BDH from E. coli DH 5alpha/pAACRII119 are similar to those of the AACRII/BDH from B. cereus YUF-4. The AACRII/BDH was considered to belong to the NAD(P)- and zinc-dependent long-chain alcohol dehydrogenase (group I ADH) family on the basis of the following distinctive characteristics: it possessed 14 strictly conserved residues of microbial group I ADH and consisted of about 350 amino acids. The enzymatic and genetic characteristics of AACRII/BDH were completely different from those of BDHs belonging to the short-chain dehydrogenase/reductase family. These findings indicated that the AACRII/BDH could be considered a new type of BDH.

  13. Caenorhabditis elegans expressing the Saccharomyces cerevisiae NADH alternative dehydrogenase Ndi1p, as a tool to identify new genes involved in complex I related diseases

    Directory of Open Access Journals (Sweden)

    Raynald eCossard

    2015-06-01

    Full Text Available Isolated complex I deficiencies are one of the most commonly observed biochemical features in patients suffering from mitochondrial disorders. In the majority of these clinical cases the molecular bases of the diseases remain unknown suggesting the involvement of unidentified factors that are critical for complex I function.The Saccharomyces cerevisiae NDI1 gene, encoding the mitochondrial internal NADH dehydrogenase was previously shown to complement a complex I deficient strain in Caenorhabitis elegans with notable improvements in reproduction, whole organism respiration. These features indicate that Ndi1p can functionally integrate the respiratory chain, allowing complex I deficiency complementation. Taking into account the Ndi1p ability to bypass complex I, we evaluate the possibility to extend the range of defects/mutations causing complex I deficiencies that can be alleviated by NDI1 expression.We report here that NDI1 expressing animals unexpectedly exhibit a slightly shortened lifespan, a reduction in the progeny and a depletion of the mitochondrial genome. However, Ndi1p is expressed and targeted to the mitochondria as a functional protein that confers rotenone resistance to those animals and without affecting their respiration rate and ATP content.We show that the severe embryonic lethality level caused by the RNAi knockdowns of complex I structural subunit encoding genes (e.g. NDUFV1, NDUFS1, NDUFS6, NDUFS8 or GRIM-19 human orthologs in wild type animals is significantly reduced in the Ndi1p expressing worm.All together these results open up the perspective to identify new genes involved in complex I function, assembly or regulation by screening an RNAi library of genes leading to embryonic lethality that should be rescued by NDI1 expression.

  14. Determination of the in vivo NAD:NADH ratio in Saccharomyces cerevisiae under anaerobic conditions, using alcohol dehydrogenase as sensor reaction.

    Science.gov (United States)

    Bekers, K M; Heijnen, J J; van Gulik, W M

    2015-08-01

    With the current quantitative metabolomics techniques, only whole-cell concentrations of NAD and NADH can be quantified. These measurements cannot provide information on the in vivo redox state of the cells, which is determined by the ratio of the free forms only. In this work we quantified free NAD:NADH ratios in yeast under anaerobic conditions, using alcohol dehydrogenase (ADH) and the lumped reaction of glyceraldehyde-3-phosphate dehydrogenase and 3-phosphoglycerate kinase as sensor reactions. We showed that, with an alternative accurate acetaldehyde determination method, based on rapid sampling, instantaneous derivatization with 2,4 diaminophenol hydrazine (DNPH) and quantification with HPLC, the ADH-catalysed oxidation of ethanol to acetaldehyde can be applied as a relatively fast and simple sensor reaction to quantify the free NAD:NADH ratio under anaerobic conditions. We evaluated the applicability of ADH as a sensor reaction in the yeast Saccharomyces cerevisiae, grown in anaerobic glucose-limited chemostats under steady-state and dynamic conditions. The results found in this study showed that the cytosolic redox status (NAD:NADH ratio) of yeast is at least one order of magnitude lower, and is thus much more reduced, under anaerobic conditions compared to aerobic glucose-limited steady-state conditions. The more reduced state of the cytosol under anaerobic conditions has major implications for (central) metabolism. Accurate determination of the free NAD:NADH ratio is therefore of importance for the unravelling of in vivo enzyme kinetics and to judge accurately the thermodynamic reversibility of each redox reaction.

  15. Mitochondrial targeting of human NADH dehydrogenase (ubiquinone flavoprotein 2 (NDUFV2 and its association with early-onset hypertrophic cardiomyopathy and encephalopathy

    Directory of Open Access Journals (Sweden)

    Kao Mou-Chieh

    2011-05-01

    Full Text Available Abstract Background NADH dehydrogenase (ubiquinone flavoprotein 2 (NDUFV2, containing one iron sulfur cluster ([2Fe-2S] binuclear cluster N1a, is one of the core nuclear-encoded subunits existing in human mitochondrial complex I. Defects in this subunit have been associated with Parkinson's disease, Alzheimer's disease, Bipolar disorder, and Schizophrenia. The aim of this study is to examine the mitochondrial targeting of NDUFV2 and dissect the pathogenetic mechanism of one human deletion mutation present in patients with early-onset hypertrophic cardiomyopathy and encephalopathy. Methods A series of deletion and point-mutated constructs with the c-myc epitope tag were generated to identify the location and sequence features of mitochondrial targeting sequence for NDUFV2 in human cells using the confocal microscopy. In addition, various lengths of the NDUFV2 N-terminal and C-terminal fragments were fused with enhanced green fluorescent protein to investigate the minimal region required for correct mitochondrial import. Finally, a deletion construct that mimicked the IVS2+5_+8delGTAA mutation in NDUFV2 gene and would eventually produce a shortened NDUFV2 lacking 19-40 residues was generated to explore the connection between human gene mutation and disease. Results We identified that the cleavage site of NDUFV2 was located around amino acid 32 of the precursor protein, and the first 22 residues of NDUFV2 were enough to function as an efficient mitochondrial targeting sequence to carry the passenger protein into mitochondria. A site-directed mutagenesis study showed that none of the single-point mutations derived from basic, hydroxylated and hydrophobic residues in the NDUFV2 presequence had a significant effect on mitochondrial targeting, while increasing number of mutations in basic and hydrophobic residues gradually decreased the mitochondrial import efficacy of the protein. The deletion mutant mimicking the human early-onset hypertrophic

  16. Mitochondrial targeting of human NADH dehydrogenase (ubiquinone) flavoprotein 2 (NDUFV2) and its association with early-onset hypertrophic cardiomyopathy and encephalopathy.

    Science.gov (United States)

    Liu, Hsin-Yu; Liao, Pin-Chao; Chuang, Kai-Tun; Kao, Mou-Chieh

    2011-05-06

    NADH dehydrogenase (ubiquinone) flavoprotein 2 (NDUFV2), containing one iron sulfur cluster ([2Fe-2S] binuclear cluster N1a), is one of the core nuclear-encoded subunits existing in human mitochondrial complex I. Defects in this subunit have been associated with Parkinson's disease, Alzheimer's disease, Bipolar disorder, and Schizophrenia. The aim of this study is to examine the mitochondrial targeting of NDUFV2 and dissect the pathogenetic mechanism of one human deletion mutation present in patients with early-onset hypertrophic cardiomyopathy and encephalopathy. A series of deletion and point-mutated constructs with the c-myc epitope tag were generated to identify the location and sequence features of mitochondrial targeting sequence for NDUFV2 in human cells using the confocal microscopy. In addition, various lengths of the NDUFV2 N-terminal and C-terminal fragments were fused with enhanced green fluorescent protein to investigate the minimal region required for correct mitochondrial import. Finally, a deletion construct that mimicked the IVS2+5_+8delGTAA mutation in NDUFV2 gene and would eventually produce a shortened NDUFV2 lacking 19-40 residues was generated to explore the connection between human gene mutation and disease. We identified that the cleavage site of NDUFV2 was located around amino acid 32 of the precursor protein, and the first 22 residues of NDUFV2 were enough to function as an efficient mitochondrial targeting sequence to carry the passenger protein into mitochondria. A site-directed mutagenesis study showed that none of the single-point mutations derived from basic, hydroxylated and hydrophobic residues in the NDUFV2 presequence had a significant effect on mitochondrial targeting, while increasing number of mutations in basic and hydrophobic residues gradually decreased the mitochondrial import efficacy of the protein. The deletion mutant mimicking the human early-onset hypertrophic cardiomyopathy and encephalopathy lacked 19-40 residues in

  17. Annotated compound data for modulators of detergent-solubilised or lipid-reconstituted respiratory type II NADH dehydrogenase activity obtained by compound library screening.

    Science.gov (United States)

    Dunn, Elyse A; Cook, Gregory M; Heikal, Adam

    2016-03-01

    The energy-generating membrane protein NADH dehydrogenase (NDH-2), a proposed antibacterial drug target (see "Inhibitors of type II NADH:menaquinone oxidoreductase represent a class of antitubercular drugs" Weinstein et al. 2005 [1]), was screened for modulators of activity in either detergent-solublised or lipid reconstituted (proteolipsome) form. Here we present an annotated list of compounds identified in a small-scale screen against NDH-2. The dataset contains information regarding the libraries screened, the identities of hit compounds and the physicochemical properties governing solubility and permeability. The implications of these data for future antibiotic discovery are discussed in our associated report, "Comparison of lipid and detergent enzyme environments for identifying inhibitors of membrane-bound energy-transducing proteins" [2].

  18. Annotated compound data for modulators of detergent-solubilised or lipid-reconstituted respiratory type II NADH dehydrogenase activity obtained by compound library screening

    Directory of Open Access Journals (Sweden)

    Elyse A. Dunn

    2016-03-01

    Full Text Available The energy-generating membrane protein NADH dehydrogenase (NDH-2, a proposed antibacterial drug target (see “Inhibitors of type II NADH:menaquinone oxidoreductase represent a class of antitubercular drugs” Weinstein et al. 2005 [1], was screened for modulators of activity in either detergent-solublised or lipid reconstituted (proteolipsome form. Here we present an annotated list of compounds identified in a small-scale screen against NDH-2. The dataset contains information regarding the libraries screened, the identities of hit compounds and the physicochemical properties governing solubility and permeability. The implications of these data for future antibiotic discovery are discussed in our associated report, “Comparison of lipid and detergent enzyme environments for identifying inhibitors of membrane-bound energy-transducing proteins” [2].

  19. Determination of the Subunit Molecular Mass and Composition of Alcohol Dehydrogenase by SDS-PAGE

    Science.gov (United States)

    Nash, Barbara T.

    2007-01-01

    SDS-PAGE is a simple, rapid technique that has many uses in biochemistry and is readily adaptable to the undergraduate laboratory. It is, however, a technique prone to several types of procedural pitfalls. This article describes the use of SDS-PAGE to determine the subunit molecular mass and composition of yeast alcohol dehydrogenase employing…

  20. Redox specificity of 2-hydroxyacid-coupled NAD(+)/NADH dehydrogenases: a study exploiting "reactive" arginine as a reporter of protein electrostatics.

    Science.gov (United States)

    Gupta, Pooja; Jairajpuri, Mohamad Aman; Durani, Susheel

    2013-01-01

    With "reactive" arginine as a kinetic reporter, 2-hydroxyacid dehydrogenases are assessed in basis of their specialization as NAD(+)-reducing or NADH-oxidizing enzymes. Specifically, M4 and H4 lactate dehydrogenases (LDHs) and cytoplasmic and mitochondrial malate dehydrogenases (MDHs) are compared to assess if their coenzyme specificity may involve electrostatics of cationic or neutral nicotinamide structure as the basis. The enzymes from diverse eukaryote and prokaryote sources thus are assessed in "reactivity" of functionally-critical arginine as a function of salt concentration and pH. Electrostatic calculations were performed on "reactive" arginines and found good correspondence with experiment. The reductive and oxidative LDHs and MDHs are assessed in their count over ionizable residues and in placement details of the residues in their structures as proteins. The variants found to be high or low in ΔpKa of "reactive" arginine are found to be also strong or weak cations that preferentially oxidize NADH (neutral nicotinamide structure) or reduce NAD(+) (cationic nicotinamide structure). The ionized groups of protein structure may thus be important to redox specificity of the enzyme on basis of electrostatic preference for the oxidized (cationic nicotinamide) or reduced (neutral nicotinamide) coenzyme. Detailed comparisons of isozymes establish that the residues contributing in their redox specificity are scrambled in structure of the reductive enzyme.

  1. Redox specificity of 2-hydroxyacid-coupled NAD(+/NADH dehydrogenases: a study exploiting "reactive" arginine as a reporter of protein electrostatics.

    Directory of Open Access Journals (Sweden)

    Pooja Gupta

    Full Text Available With "reactive" arginine as a kinetic reporter, 2-hydroxyacid dehydrogenases are assessed in basis of their specialization as NAD(+-reducing or NADH-oxidizing enzymes. Specifically, M4 and H4 lactate dehydrogenases (LDHs and cytoplasmic and mitochondrial malate dehydrogenases (MDHs are compared to assess if their coenzyme specificity may involve electrostatics of cationic or neutral nicotinamide structure as the basis. The enzymes from diverse eukaryote and prokaryote sources thus are assessed in "reactivity" of functionally-critical arginine as a function of salt concentration and pH. Electrostatic calculations were performed on "reactive" arginines and found good correspondence with experiment. The reductive and oxidative LDHs and MDHs are assessed in their count over ionizable residues and in placement details of the residues in their structures as proteins. The variants found to be high or low in ΔpKa of "reactive" arginine are found to be also strong or weak cations that preferentially oxidize NADH (neutral nicotinamide structure or reduce NAD(+ (cationic nicotinamide structure. The ionized groups of protein structure may thus be important to redox specificity of the enzyme on basis of electrostatic preference for the oxidized (cationic nicotinamide or reduced (neutral nicotinamide coenzyme. Detailed comparisons of isozymes establish that the residues contributing in their redox specificity are scrambled in structure of the reductive enzyme.

  2. Polymyxin B identified as an inhibitor of alternative NADH dehydrogenase and malate: quinone oxidoreductase from the Gram-positive bacterium Mycobacterium smegmatis.

    Science.gov (United States)

    Mogi, Tatsushi; Murase, Yoshiro; Mori, Mihoko; Shiomi, Kazuro; Omura, Satoshi; Paranagama, Madhavi P; Kita, Kiyoshi

    2009-10-01

    Tuberculosis is the leading cause of death due to a single infectious agent in the world and the emergence of multidrug-resistant strains prompted us to develop new drugs with novel targets and mechanism. Here, we screened a natural antibiotics library with Mycobacterium smegmatis membrane-bound dehydrogenases and identified polymyxin B (cationic decapeptide) and nanaomycin A (naphtoquinone derivative) as inhibitors of alternative NADH dehydrogenase [50% inhibitory concentration (IC(50)) values of 1.6 and 31 microg/ml, respectively] and malate: quinone oxidoreductase (IC(50) values of 4.2 and 49 microg/ml, respectively). Kinetic analysis on inhibition by polymyxin B showed that the primary site of action was the quinone-binding site. Because of the similarity in K(m) value for ubiquinone-1 and inhibitor sensitivity, we examined amino acid sequences of actinobacterial enzymes and found possible binding sites for L-malate and quinones. Proposed mechanisms of polymyxin B and nanaomycin A for the bacteriocidal activity were the destruction of bacterial membranes and production of reactive oxygen species, respectively, while this study revealed their inhibitory activity on bacterial membrane-bound dehydrogenases. Screening of the library with bacterial respiratory enzymes resulted in unprecedented findings, so we are hoping that continuing efforts could identify lead compounds for new drugs targeting to mycobacterial respiratory enzymes.

  3. Characterization of a NADH-dependent glutamate dehydrogenase mutant of Arabidopsis demonstrates the key role of this enzyme in root carbon and nitrogen metabolism.

    Science.gov (United States)

    Fontaine, Jean-Xavier; Tercé-Laforgue, Thérèse; Armengaud, Patrick; Clément, Gilles; Renou, Jean-Pierre; Pelletier, Sandra; Catterou, Manuella; Azzopardi, Marianne; Gibon, Yves; Lea, Peter J; Hirel, Bertrand; Dubois, Frédéric

    2012-10-01

    The role of NADH-dependent glutamate dehydrogenase (GDH) was investigated by studying the physiological impact of a complete lack of enzyme activity in an Arabidopsis thaliana plant deficient in three genes encoding the enzyme. This study was conducted following the discovery that a third GDH gene is expressed in the mitochondria of the root companion cells, where all three active GDH enzyme proteins were shown to be present. A gdh1-2-3 triple mutant was constructed and exhibited major differences from the wild type in gene transcription and metabolite concentrations, and these differences appeared to originate in the roots. By placing the gdh triple mutant under continuous darkness for several days and comparing it to the wild type, the evidence strongly suggested that the main physiological function of NADH-GDH is to provide 2-oxoglutarate for the tricarboxylic acid cycle. The differences in key metabolites of the tricarboxylic acid cycle in the triple mutant versus the wild type indicated that, through metabolic processes operating mainly in roots, there was a strong impact on amino acid accumulation, in particular alanine, γ-aminobutyrate, and aspartate in both roots and leaves. These results are discussed in relation to the possible signaling and physiological functions of the enzyme at the interface of carbon and nitrogen metabolism.

  4. Electron transfer from NADH bound to horse liver alcohol dehydrogenase (NAD+ dependent dehydrogenase): visualisation of the activity in the enzyme crystals and adsorption of formazan derivatives by these crystals.

    Science.gov (United States)

    Pacaud-Mercier, Karine; Blaghen, Mohamed; Lee, Kang Min; Tritsch, Denis; Biellmann, Jean-François

    2007-02-01

    The crystals of holoenzyme from native and cross-linked alcohol dehydrogenase exhibit electron transfer from NADH to phenazinium methosulfate (PMS), and then to the tetrazolium salt sodium 3,3'-{1-[(phenylamino)carbonyl]-3,4-tetrazolium}-bis(4-methoxy-6-nitro)benzenesulfonate (XXT). The slow dissociation of the cofactor and/or the conformational change associated can now be bypassed. The reduction product, formazan, did not diffuse out of the crystals in buffer and the crystals turned colored. In the presence of dimethyl sulfoxide or dimethoxyethane, the formazan diffused out to the solution. The reaction rates were found to be, respectively, 18% and 15% of the redox reaction rate of ethanol with cinnamaldehyde, close to the activity determined for the enzyme in solution in the presence of dimethoxyethane. The use of system PMS-tetrazolium salt is a useful tool to visualize the activity of dehydrogenases and other electron transferring systems in the crystalline state. The adsorption of formazan by the alcohol dehydrogenase crystals occurs in solution.

  5. Localization of the nucleic acid channel regulatory subunit, cytosolic malate dehydrogenase.

    Science.gov (United States)

    Hanss, Basil; Leal-Pinto, Edgar; Teixeira, Avelino; Tran, Baohuong; Lee, Chun-Hui; Henderson, Scott C; Klotman, Paul E

    2008-01-01

    NACh is a nucleic acid-conducting channel found in apical membrane of rat kidney proximal tubules. It is a heteromultimeric complex consisting of at least two proteins: a 45-kDa pore-forming subunit and a 36-kDa regulatory subunit. The regulatory subunit confers ion selectivity and influences gating kinetics. The regulatory subunit has been identified as cytosolic malate dehydrogenase (cMDH). cMDH is described in the literature as a soluble protein that is not associated with plasma membrane. Yet a role for cMDH as the regulatory subunit of NACh requires that it be present at the plasma membrane. To resolve this conflict, studies were initiated to determine whether cMDH could be found at the plasma membrane. Before performing localization studies, a suitable model system that expressed NACh was identified. A channel was identified in LLC-PK(1) cells, a line derived from pig proximal tubule, that is selective for nucleic acid and has a conductance of approximately 10 pS. It exhibits dose-dependent blockade by heparan sulfate or L-malate. These characteristics are similar to what has been reported for NACh from rat kidney and indicate that NACh is present in LLC-PK(1) cells. LLC-PK(1) cells were therefore used as a model system for immunolocalization of cMDH. Both immunofluorescence and immunoelectron microscopy demonstrated cMDH at the plasma membrane of LLC-PK(1) cells. This finding supports prior functional data that describe a role for cMDH as the regulatory subunit of NACh.

  6. Organ-specific expression of glutamate dehydrogenase (GDH) subunits in yellow lupine.

    Science.gov (United States)

    Lehmann, Teresa; Dabert, Mirosława; Nowak, Witold

    2011-07-01

    Glutamate dehydrogenase (GDH, EC 1.4.2-4) is present in yellow lupine (Lupinus luteus cv. Juno) in many isoforms. The number and banding pattern of isoenzymes varies with respect to plant organ and developmental stage. To better understand the complex nature of GDH regulation in plants, the levels of GDH transcripts, enzyme activity and isoenzyme patterns in germinating seeds and roots of yellow lupine were examined. The analysis of GDH cDNA sequences in lupine revealed three mRNA types, of which two encoded the β-GDH subunit and one encoded the α-GDH subunit (corresponding to the GDH1(GDH3) and GDH2 genes, respectively). The relative expression of GDH1 and GDH2 genes was analyzed in various lupine organs by using quantitative real-time PCR. Our results indicate that different mRNA types were differently regulated depending on organ type. Although both genes appeared to be ubiquitously expressed in all lupine tissues, the GDH1 transcripts evidently predominated over those of GDH2. Immunochemical analyses confirmed that, during embryo development, varied expression of two GDH subunits takes place. The α-GDH subunit (43kDa) predominated in the early stages of germinating seeds, while the β-GDH subunit (44kDa) was the only GDH polypeptide present in lupine roots. These results firmly support the hypothesis that isoenzyme variability of GDH in yellow lupine is associated with the varied expression of α and β subunits into the complexes of hexameric GDH forms. The presence of several isogenes of GDH in yellow lupine may explain the high number (over 20) of its molecular forms in germinating lupine. Copyright © 2011 Elsevier GmbH. All rights reserved.

  7. Evidence for Lateral Transfer of Genes Encoding Ferredoxins, Nitroreductases, NADH Oxidase, and Alcohol Dehydrogenase 3 from Anaerobic Prokaryotes to Giardia lamblia and Entamoeba histolytica

    Science.gov (United States)

    Nixon, Julie E. J.; Wang, Amy; Field, Jessica; Morrison, Hilary G.; McArthur, Andrew G.; Sogin, Mitchell L.; Loftus, Brendan J.; Samuelson, John

    2002-01-01

    Giardia lamblia and Entamoeba histolytica are amitochondriate, microaerophilic protists which use fermentation enzymes like those of bacteria to survive anaerobic conditions within the intestinal lumen. Genes encoding fermentation enzymes and related electron transport peptides (e.g., ferredoxins) in giardia organisms and amebae are hypothesized to be derived from either an ancient anaerobic eukaryote (amitochondriate fossil hypothesis), a mitochondrial endosymbiont (hydrogen hypothesis), or anaerobic bacteria (lateral transfer hypothesis). The goals here were to complete the molecular characterization of giardial and amebic fermentation enzymes and to determine the origins of the genes encoding them, when possible. A putative giardia [2Fe-2S]ferredoxin which had a hypothetical organelle-targeting sequence at its N terminus showed similarity to mitochondrial ferredoxins and the hydrogenosomal ferredoxin of Trichomonas vaginalis (another luminal protist). However, phylogenetic trees were star shaped, with weak bootstrap support, so we were unable to confirm or rule out the endosymbiotic origin of the giardia [2Fe-2S]ferredoxin gene. Putative giardial and amebic 6-kDa ferredoxins, ferredoxin-nitroreductase fusion proteins, and oxygen-insensitive nitroreductases each tentatively supported the lateral transfer hypothesis. Although there were not enough sequences to perform meaningful phylogenetic analyses, the unique common occurrence of these peptides and enzymes in giardia organisms, amebae, and the few anaerobic prokaryotes suggests the possibility of lateral transfer. In contrast, there was more robust phylogenetic evidence for the lateral transfer of G. lamblia genes encoding an NADH oxidase from a gram-positive coccus and a microbial group 3 alcohol dehydrogenase from thermoanaerobic prokaryotes. In further support of lateral transfer, the G. lamblia NADH oxidase and adh3 genes appeared to have an evolutionary history distinct from those of E. histolytica. PMID

  8. Inhibiting sperm pyruvate dehydrogenase complex and its E3 subunit, dihydrolipoamide dehydrogenase affects fertilization in Syrian hamsters.

    Directory of Open Access Journals (Sweden)

    Archana B Siva

    Full Text Available BACKGROUND/AIMS: The importance of sperm capacitation for mammalian fertilization has been confirmed in the present study via sperm metabolism. Involvement of the metabolic enzymes pyruvate dehydrogenase complex (PDHc and its E3 subunit, dihydrolipoamide dehydrogenase (DLD in hamster in vitro fertilization (IVF via in vitro sperm capacitation is being proposed through regulation of sperm intracellular lactate, pH and calcium. METHODOLOGY AND PRINCIPAL FINDINGS: Capacitated hamster spermatozoa were allowed to fertilize hamster oocytes in vitro which were then assessed for fertilization, microscopically. PDHc/DLD was inhibited by the use of the specific DLD-inhibitor, MICA (5-methoxyindole-2-carboxylic acid. Oocytes fertilized with MICA-treated (MT [and thus PDHc/DLD-inhibited] spermatozoa showed defective fertilization where 2nd polar body release and pronuclei formation were not observed. Defective fertilization was attributable to capacitation failure owing to high lactate and low intracellular pH and calcium in MT-spermatozoa during capacitation. Moreover, this defect could be overcome by alkalinizing spermatozoa, before fertilization. Increasing intracellular calcium in spermatozoa pre-IVF and in defectively-fertilized oocytes, post-fertilization rescued the arrest seen, suggesting the role of intracellular calcium from either of the gametes in fertilization. Parallel experiments carried out with control spermatozoa capacitated in medium with low extracellular pH or high lactate substantiated the necessity of optimal sperm intracellular lactate levels, intracellular pH and calcium during sperm capacitation, for proper fertilization. CONCLUSIONS: This study confirms the importance of pyruvate/lactate metabolism in capacitating spermatozoa for successful fertilization, besides revealing for the first time the importance of sperm PDHc/ DLD in fertilization, via the modulation of sperm intracellular lactate, pH and calcium during capacitation. In

  9. Inhibiting Sperm Pyruvate Dehydrogenase Complex and Its E3 Subunit, Dihydrolipoamide Dehydrogenase Affects Fertilization in Syrian Hamsters

    Science.gov (United States)

    Sailasree, Purnima; Singh, Durgesh K.; Kameshwari, Duvurri B.; Shivaji, Sisinthy

    2014-01-01

    Background/Aims The importance of sperm capacitation for mammalian fertilization has been confirmed in the present study via sperm metabolism. Involvement of the metabolic enzymes pyruvate dehydrogenase complex (PDHc) and its E3 subunit, dihydrolipoamide dehydrogenase (DLD) in hamster in vitro fertilization (IVF) via in vitro sperm capacitation is being proposed through regulation of sperm intracellular lactate, pH and calcium. Methodology and Principal Findings Capacitated hamster spermatozoa were allowed to fertilize hamster oocytes in vitro which were then assessed for fertilization, microscopically. PDHc/DLD was inhibited by the use of the specific DLD-inhibitor, MICA (5-methoxyindole-2-carboxylic acid). Oocytes fertilized with MICA-treated (MT) [and thus PDHc/DLD-inhibited] spermatozoa showed defective fertilization where 2nd polar body release and pronuclei formation were not observed. Defective fertilization was attributable to capacitation failure owing to high lactate and low intracellular pH and calcium in MT-spermatozoa during capacitation. Moreover, this defect could be overcome by alkalinizing spermatozoa, before fertilization. Increasing intracellular calcium in spermatozoa pre-IVF and in defectively-fertilized oocytes, post-fertilization rescued the arrest seen, suggesting the role of intracellular calcium from either of the gametes in fertilization. Parallel experiments carried out with control spermatozoa capacitated in medium with low extracellular pH or high lactate substantiated the necessity of optimal sperm intracellular lactate levels, intracellular pH and calcium during sperm capacitation, for proper fertilization. Conclusions This study confirms the importance of pyruvate/lactate metabolism in capacitating spermatozoa for successful fertilization, besides revealing for the first time the importance of sperm PDHc/ DLD in fertilization, via the modulation of sperm intracellular lactate, pH and calcium during capacitation. In addition, the

  10. Crystallization of the NADH-oxidizing domain of the Na{sup +}-translocating NADH:ubiquinone oxidoreductase from Vibrio cholerae

    Energy Technology Data Exchange (ETDEWEB)

    Tao, Minli [Department of Biochemistry, University of Zürich, Winterthurerstrasse 190, 8057 Zürich (Switzerland); Türk, Karin [School of Engineering and Science, International University Bremen, 28759 Bremen (Germany); Diez, Joachim [Swiss Light Source at Paul Scherrer Institut, 5232 Villigen PSI (Switzerland); Grütter, Markus G. [Department of Biochemistry, University of Zürich, Winterthurerstrasse 190, 8057 Zürich (Switzerland); Fritz, Günter, E-mail: guenter.fritz@uni-konstanz.de [Fachbereich Biologie, Universität Konstanz, Postfach M665, Universitätsstrasse 10, 78457 Konstanz (Germany); Steuber, Julia, E-mail: guenter.fritz@uni-konstanz.de [Department of Biochemistry, University of Zürich, Winterthurerstrasse 190, 8057 Zürich (Switzerland)

    2006-02-01

    The FAD domain of the NqrF subunit from the Na{sup +}-translocating NADH dehydrogenase from V. cholerae has been purified and crystallized. A complete data set was recorded at 3.1 Å. The Na{sup +}-translocating NADH:quinone oxidoreductase (Na{sup +}-NQR) from pathogenic and marine bacteria is a respiratory complex that couples the exergonic oxidation of NADH by quinone to the transport of Na{sup +} across the membrane. The NqrF subunit oxidizes NADH and transfers the electrons to other redox cofactors in the enzyme. The FAD-containing domain of NqrF has been expressed, purified and crystallized. The purified NqrF FAD domain exhibited high rates of NADH oxidation and contained stoichiometric amounts of the FAD cofactor. Initial crystallization of the flavin domain was achieved by the sitting-drop technique using a Cartesian MicroSys4000 robot. Optimization of the crystallization conditions yielded yellow hexagonal crystals with dimensions of 30 × 30 × 70 µm. The protein mainly crystallizes in long hexagonal needles with a diameter of up to 30 µm. Crystals diffract to 2.8 Å and belong to space group P622, with unit-cell parameters a = b = 145.3, c = 90.2 Å, α = β = 90, γ = 120°.

  11. NADH:ubiquinone reductase and succinate dehydrogenase activity in the liver of rats with acetaminophen-induced toxic hepatitis on the background of alimentary protein deficiency

    Directory of Open Access Journals (Sweden)

    G. P. Kopylchuk

    2015-02-01

    Full Text Available The ratio between the redox forms of the nicotinamide coenzymes and key enzymatic activity of the I and II respiratory chain complexes in the liver cells mitochondria of rats with acetaminophen-induced hepatitis under the conditions of alimentary deprivation of protein was studied. It was estimated, that under the conditions of acute acetaminophen-induced hepatitis of rats kept on a low-protein diet during 4 weeks a significant decrease of the NADH:ubiquinone reductase and succinate dehydrogenase activity with simultaneous increase of the ratio between redox forms of the nicotinamide coenzymes (NAD+/NADН is observed compared to the same indices in the liver cells of animals with experimental hepatitis kept on the ration balanced by all nutrients. Results of research may become basic ones for the biochemical rationale for the approaches directed to the correction and elimination of the consequences­ of energy exchange in the toxic hepatitis, induced on the background of protein deficiency.

  12. Reduced levels of NADH-dependent glutamate dehydrogenase decrease the glutamate content of ripe tomato fruit but have no effect on green fruit or leaves.

    Science.gov (United States)

    Ferraro, Gisela; D'Angelo, Matilde; Sulpice, Ronan; Stitt, Mark; Valle, Estela M

    2015-06-01

    Glutamate (Glu) is a taste enhancer that contributes to the characteristic flavour of foods. In fruit of tomato (Solanum lycopersicum L.), the Glu content increases dramatically during the ripening process, becoming the most abundant free amino acid when the fruit become red. There is also a concomitant increase in NADH-dependent glutamate dehydrogenase (GDH) activity during the ripening transition. This enzyme is located in the mitochondria and catalyses the reversible amination of 2-oxoglutarate to Glu. To investigate the potential effect of GDH on Glu metabolism, the abundance of GDH was altered by artificial microRNA technology. Efficient silencing of all the endogenous SlGDH genes was achieved, leading to a dramatic decrease in total GDH activity. This decrease in GDH activity did not lead to any clear morphological or metabolic phenotype in leaves or green fruit. However, red fruit on the transgenic plants showed markedly reduced levels of Glu and a large increase in aspartate, glucose and fructose content in comparison to wild-type fruit. These results suggest that GDH is involved in the synthesis of Glu in tomato fruit during the ripening processes. This contrasts with the biological role ascribed to GDH in many other tissues and species. Overall, these findings suggest that GDH has a major effect on the control of metabolic composition during tomato fruit ripening, but not at other stages of development.

  13. Host cell and expression engineering for development of an E. coli ketoreductase catalyst: Enhancement of formate dehydrogenase activity for regeneration of NADH

    Directory of Open Access Journals (Sweden)

    Mädje Katharina

    2012-01-01

    Full Text Available Abstract Background Enzymatic NADH or NADPH-dependent reduction is a widely applied approach for the synthesis of optically active organic compounds. The overall biocatalytic conversion usually involves in situ regeneration of the expensive NAD(PH. Oxidation of formate to carbon dioxide, catalyzed by formate dehydrogenase (EC 1.2.1.2; FDH, presents an almost ideal process solution for coenzyme regeneration that has been well established for NADH. Because isolated FDH is relatively unstable under a range of process conditions, whole cells often constitute the preferred form of the biocatalyst, combining the advantage of enzyme protection in the cellular environment with ease of enzyme production. However, the most prominent FDH used in biotransformations, the enzyme from the yeast Candida boidinii, is usually expressed in limiting amounts of activity in the prime host for whole cell biocatalysis, Escherichia coli. We therefore performed expression engineering with the aim of enhancing FDH activity in an E. coli ketoreductase catalyst. The benefit resulting from improved NADH regeneration capacity is demonstrated in two transformations of technological relevance: xylose conversion into xylitol, and synthesis of (S-1-(2-chlorophenylethanol from o-chloroacetophenone. Results As compared to individual expression of C. boidinii FDH in E. coli BL21 (DE3 that gave an intracellular enzyme activity of 400 units/gCDW, co-expression of the FDH with the ketoreductase (Candida tenuis xylose reductase; XR resulted in a substantial decline in FDH activity. The remaining FDH activity of only 85 U/gCDW was strongly limiting the overall catalytic activity of the whole cell system. Combined effects from increase in FDH gene copy number, supply of rare tRNAs in a Rosetta strain of E. coli, dampened expression of the ketoreductase, and induction at low temperature (18°C brought up the FDH activity threefold to a level of 250 U/gCDW while reducing the XR activity by

  14. A novel mutation in the succinate dehydrogenase subunit D gene in siblings with the hereditary paraganglioma-pheochromocytoma syndrome.

    Science.gov (United States)

    Prasad, Chaithra; Oakley, Gerard J; Yip, Linwah; Coyne, Christopher; Rangaswamy, Balasubramanya; Dixit, Sanjay B

    2014-01-01

    Germline mutations in the succinate dehydrogenase complex subunit D gene are now known to be associated with hereditary paraganglioma-pheochromocytoma syndromes. Since the initial succinate dehydrogenase complex subunit D gene mutation was identified about a decade ago, more than 131 unique variants have been reported. We report the case of two siblings presenting with multiple paragangliomas and pheochromocytomas; they were both found to carry a mutation in the succinate dehydrogenase complex subunit D gene involving a substitution of thymine to guanine at nucleotide 236 in exon 3. This particular mutation of the succinate dehydrogenase complex subunit D gene has only been reported in one previous patient in Japan; this is, therefore, the first report of this pathogenic mutation in siblings and the first report of this mutation in North America. With continued screening of more individuals, we will be able to create a robust mutation database that can help us understand disease patterns associated with particular variants and may be a starting point in the development of new therapies for familial paraganglioma syndromes.

  15. A novel mutation in the succinate dehydrogenase subunit D gene in siblings with the hereditary paraganglioma–pheochromocytoma syndrome

    Directory of Open Access Journals (Sweden)

    Chaithra Prasad

    2014-10-01

    Full Text Available Germline mutations in the succinate dehydrogenase complex subunit D gene are now known to be associated with hereditary paraganglioma–pheochromocytoma syndromes. Since the initial succinate dehydrogenase complex subunit D gene mutation was identified about a decade ago, more than 131 unique variants have been reported. We report the case of two siblings presenting with multiple paragangliomas and pheochromocytomas; they were both found to carry a mutation in the succinate dehydrogenase complex subunit D gene involving a substitution of thymine to guanine at nucleotide 236 in exon 3. This particular mutation of the succinate dehydrogenase complex subunit D gene has only been reported in one previous patient in Japan; this is, therefore, the first report of this pathogenic mutation in siblings and the first report of this mutation in North America. With continued screening of more individuals, we will be able to create a robust mutation database that can help us understand disease patterns associated with particular variants and may be a starting point in the development of new therapies for familial paraganglioma syndromes.

  16. A novel mutation in the succinate dehydrogenase subunit D gene in siblings with the hereditary paraganglioma–pheochromocytoma syndrome

    Science.gov (United States)

    Oakley, Gerard J; Yip, Linwah; Coyne, Christopher; Rangaswamy, Balasubramanya; Dixit, Sanjay B

    2014-01-01

    Germline mutations in the succinate dehydrogenase complex subunit D gene are now known to be associated with hereditary paraganglioma–pheochromocytoma syndromes. Since the initial succinate dehydrogenase complex subunit D gene mutation was identified about a decade ago, more than 131 unique variants have been reported. We report the case of two siblings presenting with multiple paragangliomas and pheochromocytomas; they were both found to carry a mutation in the succinate dehydrogenase complex subunit D gene involving a substitution of thymine to guanine at nucleotide 236 in exon 3. This particular mutation of the succinate dehydrogenase complex subunit D gene has only been reported in one previous patient in Japan; this is, therefore, the first report of this pathogenic mutation in siblings and the first report of this mutation in North America. With continued screening of more individuals, we will be able to create a robust mutation database that can help us understand disease patterns associated with particular variants and may be a starting point in the development of new therapies for familial paraganglioma syndromes. PMID:27489656

  17. Frequent germ-line succinate dehydrogenase subunit D gene mutations in patients with apparently sporadic parasympathetic paraganglioma

    NARCIS (Netherlands)

    H. Dannenberg (Hilde); W.N.M. Dinjens (Winand); M. Abbou; H. van Urk (Hero); B.K. Pauw; D. Mouwen; W.J. Mooi (Wolter); R.R. de Krijger (Ronald)

    2002-01-01

    textabstractPURPOSE: Recently, familial paraganglioma (PGL) was shown to be caused bymutations in the gene encoding succinate dehydrogenase subunit D (SDHD). However, the prevalence of SDHD mutations in apparently sporadic PGL is unknown. We studied the frequency and spectrum of ge

  18. FMN is covalently attached to a threonine residue in the NqrB and NqrC subunits of Na(+)-translocating NADH-quinone reductase from Vibrio alginolyticus.

    Science.gov (United States)

    Hayashi, M; Nakayama, Y; Yasui, M; Maeda, M; Furuishi, K; Unemoto, T

    2001-01-12

    The Na(+)-translocating NADH-quinone reductase (NQR) from Vibrio alginolyticus is composed of six subunits (NqrA to NqrF). We previously demonstrated that both NqrB and NqrC subunits contain a flavin cofactor covalently attached to a threonine residue. Fluorescent peptide fragments derived from the NqrB and NqrC subunits were applied to a matrix-assisted laser desorption ionization time-of-flight mass spectrometer, and covalently attached flavin was identified as FMN in both subunits. From post-source decay fragmentation analysis, it was concluded that FMN is attached by a phosphate group to Thr-235 in the NqrB subunit and to Thr-223 in the NqrC subunit. The phosphoester binding of FMN to a threonine residue reported here is a new type of flavin attachment to a polypeptide.

  19. Immobilisation and characterisation of biocatalytic co-factor recycling enzymes, glucose dehydrogenase and NADH oxidase, on aldehyde functional ReSyn™ polymer microspheres.

    Science.gov (United States)

    Twala, Busisiwe V; Sewell, B Trevor; Jordaan, Justin

    2012-05-10

    The use of enzymes in industrial applications is limited by their instability, cost and difficulty in their recovery and re-use. Immobilisation is a technique which has been shown to alleviate these limitations in biocatalysis. Here we describe the immobilisation of two biocatalytically relevant co-factor recycling enzymes, glucose dehydrogenase (GDH) and NADH oxidase (NOD) on aldehyde functional ReSyn™ polymer microspheres with varying functional group densities. The successful immobilisation of the enzymes on this new high capacity microsphere technology resulted in the maintenance of activity of ∼40% for GDH and a maximum of 15.4% for NOD. The microsphere variant with highest functional group density of ∼3500 μmol g⁻¹ displayed the highest specific activity for the immobilisation of both enzymes at 33.22 U mg⁻¹ and 6.75 U mg⁻¹ for GDH and NOD with respective loading capacities of 51% (0.51 mg mg⁻¹) and 129% (1.29 mg mg⁻¹). The immobilised GDH further displayed improved activity in the acidic pH range. Both enzymes displayed improved pH and thermal stability with the most pronounced thermal stability for GDH displayed on ReSyn™ A during temperature incubation at 65 °C with a 13.59 fold increase, and NOD with a 2.25-fold improvement at 45 °C on the same microsphere variant. An important finding is the suitability of the microspheres for stabilisation of the multimeric protein GDH.

  20. Structural and functional investigation of flavin binding center of the NqrC subunit of sodium-translocating NADH:quinone oxidoreductase from Vibrio harveyi.

    Directory of Open Access Journals (Sweden)

    Valentin Borshchevskiy

    Full Text Available Na+-translocating NADH:quinone oxidoreductase (NQR is a redox-driven sodium pump operating in the respiratory chain of various bacteria, including pathogenic species. The enzyme has a unique set of redox active prosthetic groups, which includes two covalently bound flavin mononucleotide (FMN residues attached to threonine residues in subunits NqrB and NqrC. The reason of FMN covalent bonding in the subunits has not been established yet. In the current work, binding of free FMN to the apo-form of NqrC from Vibrio harveyi was studied showing very low affinity of NqrC to FMN in the absence of its covalent bonding. To study structural aspects of flavin binding in NqrC, its holo-form was crystallized and its 3D structure was solved at 1.56 Å resolution. It was found that the isoalloxazine moiety of the FMN residue is buried in a hydrophobic cavity and that its pyrimidine ring is squeezed between hydrophobic amino acid residues while its benzene ring is extended from the protein surroundings. This structure of the flavin-binding pocket appears to provide flexibility of the benzene ring, which can help the FMN residue to take the bended conformation and thus to stabilize the one-electron reduced form of the prosthetic group. These properties may also lead to relatively weak noncovalent binding of the flavin. This fact along with periplasmic location of the FMN-binding domains in the vast majority of NqrC-like proteins may explain the necessity of the covalent bonding of this prosthetic group to prevent its loss to the external medium.

  1. Structural and functional investigation of flavin binding center of the NqrC subunit of sodium-translocating NADH:quinone oxidoreductase from Vibrio harveyi.

    Science.gov (United States)

    Borshchevskiy, Valentin; Round, Ekaterina; Bertsova, Yulia; Polovinkin, Vitaly; Gushchin, Ivan; Ishchenko, Andrii; Kovalev, Kirill; Mishin, Alexey; Kachalova, Galina; Popov, Alexander; Bogachev, Alexander; Gordeliy, Valentin

    2015-01-01

    Na+-translocating NADH:quinone oxidoreductase (NQR) is a redox-driven sodium pump operating in the respiratory chain of various bacteria, including pathogenic species. The enzyme has a unique set of redox active prosthetic groups, which includes two covalently bound flavin mononucleotide (FMN) residues attached to threonine residues in subunits NqrB and NqrC. The reason of FMN covalent bonding in the subunits has not been established yet. In the current work, binding of free FMN to the apo-form of NqrC from Vibrio harveyi was studied showing very low affinity of NqrC to FMN in the absence of its covalent bonding. To study structural aspects of flavin binding in NqrC, its holo-form was crystallized and its 3D structure was solved at 1.56 Å resolution. It was found that the isoalloxazine moiety of the FMN residue is buried in a hydrophobic cavity and that its pyrimidine ring is squeezed between hydrophobic amino acid residues while its benzene ring is extended from the protein surroundings. This structure of the flavin-binding pocket appears to provide flexibility of the benzene ring, which can help the FMN residue to take the bended conformation and thus to stabilize the one-electron reduced form of the prosthetic group. These properties may also lead to relatively weak noncovalent binding of the flavin. This fact along with periplasmic location of the FMN-binding domains in the vast majority of NqrC-like proteins may explain the necessity of the covalent bonding of this prosthetic group to prevent its loss to the external medium.

  2. An Fe-S cluster in the conserved Cys-rich region in the catalytic subunit of FAD-dependent dehydrogenase complexes.

    Science.gov (United States)

    Shiota, Masaki; Yamazaki, Tomohiko; Yoshimatsu, Keiichi; Kojima, Katsuhiro; Tsugawa, Wakako; Ferri, Stefano; Sode, Koji

    2016-12-01

    Several bacterial flavin adenine dinucleotide (FAD)-harboring dehydrogenase complexes comprise three distinct subunits: a catalytic subunit with FAD, a cytochrome c subunit containing three hemes, and a small subunit. Owing to the cytochrome c subunit, these dehydrogenase complexes have the potential to transfer electrons directly to an electrode. Despite various electrochemical applications and engineering studies of FAD-dependent dehydrogenase complexes, the intra/inter-molecular electron transfer pathway has not yet been revealed. In this study, we focused on the conserved Cys-rich region in the catalytic subunits using the catalytic subunit of FAD dependent glucose dehydrogenase complex (FADGDH) as a model, and site-directed mutagenesis and electron paramagnetic resonance (EPR) were performed. By co-expressing a hitch-hiker protein (γ-subunit) and a catalytic subunit (α-subunit), FADGDH γα complexes were prepared, and the properties of the catalytic subunit of both wild type and mutant FADGDHs were investigated. Substitution of the conserved Cys residues with Ser resulted in the loss of dye-mediated glucose dehydrogenase activity. ICP-AEM and EPR analyses of the wild-type FADGDH catalytic subunit revealed the presence of a 3Fe-4S-type iron-sulfur cluster, whereas none of the Ser-substituted mutants showed the EPR spectrum characteristic for this cluster. The results suggested that three Cys residues in the Cys-rich region constitute an iron-sulfur cluster that may play an important role in the electron transfer from FAD (intra-molecular) to the multi-heme cytochrome c subunit (inter-molecular) electron transfer pathway. These features appear to be conserved in the other three-subunit dehydrogenases having an FAD cofactor.

  3. Succinate Dehydrogenase B Subunit Immunohistochemical Expression Predicts Aggressiveness in Well Differentiated Neuroendocrine Tumors of the Ileum

    Energy Technology Data Exchange (ETDEWEB)

    Milione, Massimo [Department of Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan I-20133 (Italy); Pusceddu, Sara [Department of Medical Oncology, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan I-20133 (Italy); Gasparini, Patrizia [Molecular Cytogenetics Unit, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan I-20133 (Italy); Melotti, Flavia [Department of Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan I-20133 (Italy); Maisonneuve, Patrick [Division of Epidemiology and Biostatistics, European Institute of Oncology, Milan 20141 (Italy); Mazzaferro, Vincenzo [Division of Gastrointestinal Surgery and Liver Transplantation, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan I-20133 (Italy); Braud, Filippo G. de [Department of Medical Oncology, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan I-20133 (Italy); Pelosi, Giuseppe, E-mail: giuseppe.pelosi@unimi.it [Department of Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan I-20133 (Italy); Department of Medicine, Surgery and Dentistry, Università degli Studi, Facoltà di Medicina, Milan 20122 (Italy)

    2012-08-16

    Immunohistochemical loss of the succinate dehydrogenase subunit B (SDHB) has recently been reported as a surrogate biomarker of malignancy in sporadic and familial pheocromocytomas and paragangliomas through the activation of hypoxia pathways. However, data on the prevalence and the clinical implications of SDHB immunoreactivity in ileal neuroendocrine tumors are still lacking. Thirty-one consecutive, advanced primary midgut neuroendocrine tumors and related lymph node or liver metastases from 24 males and seven females were immunohistochemically assessed for SDHB. All patients were G1 tumors (Ki-67 labeling index ≤2%). SDHB immunohistochemistry results were expressed as immunostaining intensity and scored as low or strong according to the internal control represented by normal intestinal cells. Strong positivity for SDHB, with granular cytoplasmatic reactivity, was found in 77% of primary tumors (T), whilst low SDHB expression was detected in 90% of metastases (M). The combined analysis (T+M) confirmed the loss of SDHB expression in 82% of metastases compared to 18% of primary tumors. SDHB expression was inversely correlated with Ki-67 labeling index, which accounted for 1.54% in metastastic sites and 0.7% in primary tumors. A correlation between SDHB expression loss, increased Ki-67 labeling index and biological aggressiveness was shown in advanced midgut neuroendocrine tumors, suggesting a role of tumor suppressor gene.

  4. Decrease in nicotinamide adenine dinucleotide dehydrogenase is related to skin pigmentation.

    Science.gov (United States)

    Nakama, Mitsuo; Murakami, Yuhko; Tanaka, Hiroshi; Nakata, Satoru

    2012-03-01

    Skin pigmentation is caused by various physical and chemical factors. It might also be influenced by changes in the physiological function of skin with aging. Nicotinamide adenine dinucleotide (NADH) dehydrogenase is an enzyme related to the mitochondrial electron transport system and plays a key role in cellular energy production. It has been reported that the functional decrease in this system causes Parkinson's disease. Another study reports that the amount of NADH dehydrogenase in heart and skeletal muscle decreases with aging. A similar decrease in the skin would probably affect its physiological function. However, no reports have examined the age-related change in levels of NADH dehydrogenase in human skin. In this study, we investigated this change and its effect on skin pigmentation using cultured human epidermal keratinocytes. The mRNA expression of NDUFA1, NDUFB7, and NDUFS2, subunits of NADH dehydrogenase, and its activity were significantly decreased in late passage keratinocytes compared to early passage cells. Conversely, the mRNA expression of melanocyte-stimulating cytokines, interleukin-1 alpha and endothelin 1, was increased in late passage cells. On the other hand, the inhibition of NADH dehydrogenase upregulated the mRNA expression of melanocyte-stimulating cytokines. Moreover, the level of NDUFB7 mRNA was lower in pigmented than in nonpigmented regions of skin in vivo. These results suggest the decrease in NADH dehydrogenase with aging to be involved in skin pigmentation.

  5. Successful chemotherapy of hepatic metastases in a case of succinate dehydrogenase subunit B-related paraganglioma.

    Science.gov (United States)

    He, J; Makey, D; Fojo, T; Adams, K T; Havekes, B; Eisenhofer, G; Sullivan, P; Lai, E W; Pacak, K

    2009-10-01

    Compared to other familial pheochromocytoma/paragangliomas (PHEO/PGLs), the succinate dehydrogenase subunit B (SDHB)-related PHEO/PGLs often present with aggressive and rapidly growing metastatic lesions. Currently, there is no proven effective treatment for malignant PHEO/PGLs. Here, we present a 35-year-old white man with primary malignant abdominal extra-adrenal 11 cm paraganglioma underwent surgical successful resection. But 6 months later, he developed extensive bone, liver, and lymph nodes metastasis, which were demonstrated by computed tomography scan and the (18)F-fluorodeoxyglucose positron emission tomography. However, his (123)I-metaiodobenzylguanidine scintigraphy was negative; therefore, the cyclophosphamide, vincristine, and dacarbazine (CVD) combination chemotherapy was initiated. The combination chemotherapy was very effective showing 80% overall reduction in the liver lesions and 75% overall reduction in the retroperitoneal mass and adenopathy, and normalization of plasma catecholamine and metanephrine levels. However, plasma levels of dopamine (DA) and methoxytyramine (MTY) were only partially affected and remained consistently elevated throughout the remaining period of follow-up evaluation. Genetic testing revealed an SDHB gene mutation. Here, we present an SDHB-related PHEO/PGL patient with extensive tumor burden, numerous organ lesions, and rapidly growing tumors, which responded extremely well to CVD therapy. We conclude patients with SDHB-related PHEO/PGLs can be particularly sensitive to CVD chemotherapy and may have an excellent outcome if this therapy is used and continued on periodic basis. The data in this patient also illustrate the importance of measuring plasma levels of DA and MTY to provide a more complete and accurate assessment of the biochemical response to therapy than provided by measurements restricted to other catecholamines and O-methylated metabolites.

  6. Chorion peroxidase-mediated NADH/O2 oxidoreduction cooperated by chorion malate dehydrogenase-catalyzed NADH production: a feasible pathway leading to H2O2 formation during chorion hardening in Aedes aegypti mosquitoes

    OpenAIRE

    Han, Qian; Li, Guoyu; Li, Jianyong

    2000-01-01

    A specific chorion peroxidase is present in Aedes aegypti and this enzyme is responsible for catalyzing chorion protein cross-linking through dityrosine formation during chorion hardening. Peroxidase-mediated dityrosine cross-linking requires H2O2, and this study discusses the possible involvement of the chorion peroxidase in H2O2 formation by mediating NADH/O2 oxidoreduction during chorion hardening in A. aegypti eggs. Our data show that mosquito chorion peroxidase is able to catalyze pH-dep...

  7. Aspartic Acid 397 in Subunit B of the Na+-pumping NADH:Quinone Oxidoreductase from Vibrio cholerae Forms Part of a Sodium-binding Site, Is Involved in Cation Selectivity, and Affects Cation-binding Site Cooperativity

    Science.gov (United States)

    Shea, Michael E.; Juárez, Oscar; Cho, Jonathan; Barquera, Blanca

    2013-01-01

    The Na+-pumping NADH:quinone complex is found in Vibrio cholerae and other marine and pathogenic bacteria. NADH:ubiquinone oxidoreductase oxidizes NADH and reduces ubiquinone, using the free energy released by this reaction to pump sodium ions across the cell membrane. In a previous report, a conserved aspartic acid residue in the NqrB subunit at position 397, located in the cytosolic face of this protein, was proposed to be involved in the capture of sodium. Here, we studied the role of this residue through the characterization of mutant enzymes in which this aspartic acid was substituted by other residues that change charge and size, such as arginine, serine, lysine, glutamic acid, and cysteine. Our results indicate that NqrB-Asp-397 forms part of one of the at least two sodium-binding sites and that both size and charge at this position are critical for the function of the enzyme. Moreover, we demonstrate that this residue is involved in cation selectivity, has a critical role in the communication between sodium-binding sites, by promoting cooperativity, and controls the electron transfer step involved in sodium uptake (2Fe-2S → FMNC). PMID:24030824

  8. The E1 beta-subunit of pyruvate dehydrogenase is surface-expressed in Lactobacillus plantarum and binds fibronectin.

    Science.gov (United States)

    Vastano, Valeria; Salzillo, Marzia; Siciliano, Rosa A; Muscariello, Lidia; Sacco, Margherita; Marasco, Rosangela

    2014-01-01

    Lactobacillus plantarum is among the species with a probiotic activity. Adhesion of probiotic bacteria to host tissues is an important principle for strain selection, because it represents a crucial step in the colonization process of either pathogens or commensals. Most bacterial adhesins are proteins, and a major target for them is fibronectin, an extracellular matrix glycoprotein. In this study we demonstrate that PDHB, a component of the pyruvate dehydrogenase complex, is a factor contributing to fibronectin-binding in L. plantarum LM3. By means of fibronectin overlay immunoblotting assay, we identified a L. plantarum LM3 surface protein with apparent molecular mass of 35 kDa. Mass spectrometric analysis shows that this protein is the pyruvate dehydrogenase E1 beta-subunit (PDHB). The corresponding pdhB gene is located in a 4-gene cluster encoding pyruvate dehydrogenase. In LM3-B1, carrying a null mutation in pdhB, the 35 kDa adhesin was not anymore detectable by immunoblotting assay. Nevertheless, the pdhB null mutation did not abolish pdhA, pdhC, and pdhD transcription in LM3-B1. By adhesion assays, we show that LM3-B1 cells bind to immobilized fibronectin less efficiently than wild type cells. Moreover, we show that pdhB expression is negatively regulated by the CcpA protein and is induced by bile.

  9. Subunits of the Pyruvate Dehydrogenase Cluster of Mycoplasma pneumoniae Are Surface-Displayed Proteins that Bind and Activate Human Plasminogen.

    Directory of Open Access Journals (Sweden)

    Anne Gründel

    Full Text Available The dual role of glycolytic enzymes in cytosol-located metabolic processes and in cell surface-mediated functions with an influence on virulence is described for various micro-organisms. Cell wall-less bacteria of the class Mollicutes including the common human pathogen Mycoplasma pneumoniae possess a reduced genome limiting the repertoire of virulence factors and metabolic pathways. After the initial contact of bacteria with cells of the respiratory epithelium via a specialized complex of adhesins and release of cell-damaging factors, surface-displayed glycolytic enzymes may facilitate the further interaction between host and microbe. In this study, we described detection of the four subunits of pyruvate dehydrogenase complex (PDHA-D among the cytosolic and membrane-associated proteins of M. pneumoniae. Subunits of PDH were cloned, expressed and purified to produce specific polyclonal guinea pig antisera. Using colony blotting, fractionation of total proteins and immunofluorescence experiments, the surface localization of PDHA-C was demonstrated. All recombinant PDH subunits are able to bind to HeLa cells and human plasminogen. These interactions can be specifically blocked by the corresponding polyclonal antisera. In addition, an influence of ionic interactions on PDHC-binding to plasminogen as well as of lysine residues on the association of PDHA-D with plasminogen was confirmed. The PDHB subunit was shown to activate plasminogen and the PDHB-plasminogen complex induces degradation of human fibrinogen. Hence, our data indicate that the surface-associated PDH subunits might play a role in the pathogenesis of M. pneumoniae infections by interaction with human plasminogen.

  10. The conformational changes induced by ubiquinone binding in the Na+-pumping NADH:ubiquinone oxidoreductase (Na+-NQR) are kinetically controlled by conserved glycines 140 and 141 of the NqrB subunit.

    Science.gov (United States)

    Strickland, Madeleine; Juárez, Oscar; Neehaul, Yashvin; Cook, Darcie A; Barquera, Blanca; Hellwig, Petra

    2014-08-22

    Na(+)-pumping NADH:ubiquinone oxidoreductase (Na(+)-NQR) is responsible for maintaining a sodium gradient across the inner bacterial membrane. This respiratory enzyme, which couples sodium pumping to the electron transfer between NADH and ubiquinone, is not present in eukaryotes and as such could be a target for antibiotics. In this paper it is shown that the site of ubiquinone reduction is conformationally coupled to the NqrB subunit, which also hosts the final cofactor in the electron transport chain, riboflavin. Previous work showed that mutations in conserved NqrB glycine residues 140 and 141 affect ubiquinone reduction and the proper functioning of the sodium pump. Surprisingly, these mutants did not affect the dissociation constant of ubiquinone or its analog HQNO (2-n-heptyl-4-hydroxyquinoline N-oxide) from Na(+)-NQR, which indicates that these residues do not participate directly in the ubiquinone binding site but probably control its accessibility. Indeed, redox-induced difference spectroscopy showed that these mutations prevented the conformational change involved in ubiquinone binding but did not modify the signals corresponding to bound ubiquinone. Moreover, data are presented that demonstrate the NqrA subunit is able to bind ubiquinone but with a low non-catalytically relevant affinity. It is also suggested that Na(+)-NQR contains a single catalytic ubiquinone binding site and a second site that can bind ubiquinone but is not active.

  11. Efficient whole-cell biocatalyst for acetoin production with NAD+ regeneration system through homologous co-expression of 2,3-butanediol dehydrogenase and NADH oxidase in engineered Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Teng Bao

    Full Text Available Acetoin (3-hydroxy-2-butanone, an extensively-used food spice and bio-based platform chemical, is usually produced by chemical synthesis methods. With increasingly requirement of food security and environmental protection, bio-fermentation of acetoin by microorganisms has a great promising market. However, through metabolic engineering strategies, the mixed acid-butanediol fermentation metabolizes a certain portion of substrate to the by-products of organic acids such as lactic acid and acetic acid, which causes energy cost and increases the difficulty of product purification in downstream processes. In this work, due to the high efficiency of enzymatic reaction and excellent selectivity, a strategy for efficiently converting 2,3-butandiol to acetoin using whole-cell biocatalyst by engineered Bacillus subtilis is proposed. In this process, NAD+ plays a significant role on 2,3-butanediol and acetoin distribution, so the NADH oxidase and 2,3-butanediol dehydrogenase both from B. subtilis are co-expressed in B. subtilis 168 to construct an NAD+ regeneration system, which forces dramatic decrease of the intracellular NADH concentration (1.6 fold and NADH/NAD+ ratio (2.2 fold. By optimization of the enzymatic reaction and applying repeated batch conversion, the whole-cell biocatalyst efficiently produced 91.8 g/L acetoin with a productivity of 2.30 g/(L·h, which was the highest record ever reported by biocatalysis. This work indicated that manipulation of the intracellular cofactor levels was more effective than the strategy of enhancing enzyme activity, and the bioprocess for NAD+ regeneration may also be a useful way for improving the productivity of NAD+-dependent chemistry-based products.

  12. 重组NADH氧化酶对乳酸脱氢酶乳酸氧化活性的影响%Effects of Recombinant NADH Oxidase on the Lactate Oxidation Activity of Lactate Dehydrogenase

    Institute of Scientific and Technical Information of China (English)

    赵蕊; 霍贵成

    2013-01-01

    [目的]考察当存在其他利用NADH途径时,发酵型乳酸脱氢酶(lactate dehydrogenase,LDH)催化乳酸氧化能力的改变.[方法]PCR扩增乳酸乳球菌(Lactococcus lactis,L.lactis)中生成H2O的NADH氧化酶基因noxE,将其连接至表达载体并在大肠杆菌中过量表达;对亲和纯化的产物进行SDS-PAGE分析、光谱扫描和活性测定,考察纯化产物是否具有生物学活性;以2,4-二硝基苯肼法测定乳酸脱氢酶的乳酸氧化活性,考察添加NoxE重组蛋白对其活性的影响.[结果]重组NoxE蛋白是种黄素蛋白,具明显的生物学活性,说明noxE表达载体构建成功;添加NoxE后,LDH的乳酸氧化活性提高了3.84倍.[结论]在NADH经呼吸链代谢掉的生理条件下,LDH催化乳酸氧化的能力会明显提高.%[ Objective] To compare the lactate oxidation activity of lactate dehydrogenase (LDH) in the presence and absence of another NADH utilization pathway. [Method] The H2O-producing NADH oxidase gene (noxE) was cloned by PCR from Lactococcws lactis genome, ligated into the expression vector and expressed in E. coli. After affinity purification, the recombinant protein was analyzed by SDS-PAGE, UV-vis absorption spectrum and determination of enzyme activity. 2,4-Dinitrophenylhydrazine was used to evaluate the effect of NoxE addition on the lactate oxidation activity of LDH. [Result]NoxE was purified as a flavin protein with significant activity. When NoxE was added, the lactate oxidation activity of LDH was increased 3.84-fold. [ Conclusion]The lactate oxidation capacity of LDH will be significantly increased under physical conditions where NADH can be consumed via respiration chain.

  13. Clinical presentations, biochemical phenotypes, and genotype-phenotype correlations in patients with succinate dehydrogenase subunit B-associated pheochromocytomas and paragangliomas.

    NARCIS (Netherlands)

    Timmers, H.J.L.M.; Kozupa, A.; Eisenhofer, G.; Raygada, M.; Adams, K.T.; Solis, D.; Lenders, J.W.M.; Pacak, K.

    2007-01-01

    CONTEXT: Mutations of the gene encoding succinate dehydrogenase subunit B (SDHB) predispose to malignant paraganglioma (PGL). Recognition of the SDHB phenotype in apparently sporadic PGL directs appropriate treatment and family screening. OBJECTIVE: The objective of the study was to assess mutation-

  14. CLONING, SEQUENCING AND EXPRESSION STUDIES OF THE GENES ENCODING AMICYANIN AND THE BETA-SUBUNIT OF METHYLAMINE DEHYDROGENASE FROM THIOBACILLUS-VERSUTUS

    NARCIS (Netherlands)

    UBBINK, M; VANKLEEF, MAG; KLEINJAN, DJ; HOITINK, CWG; HUITEMA, F; BEINTEMA, JJ; DUINE, JA; CANTERS, GW

    1991-01-01

    The genes encoding amicyanin and the beta-subunit of methylamine dehydrogenase (MADH) from Thiobacillus versutus have been cloned and sequenced. The organization of these genes makes it likely that they are coordinately expressed and it supports earlier findings that the blue copper protein amicyani

  15. The structure of apo human glutamate dehydrogenase details subunit communication and allostery.

    Science.gov (United States)

    Smith, Thomas J; Schmidt, Timothy; Fang, Jie; Wu, Jane; Siuzdak, Gary; Stanley, Charles A

    2002-05-01

    The structure of human glutamate dehydrogenase (GDH) has been determined in the absence of active site and regulatory ligands. Compared to the structures of bovine GDH that were complexed with coenzyme and substrate, the NAD binding domain is rotated away from the glutamate-binding domain. The electron density of this domain is more disordered the further it is from the pivot helix. Mass spectrometry results suggest that this is likely due to the apo form being more dynamic than the closed form. The antenna undergoes significant conformational changes as the catalytic cleft opens. The ascending helix in the antenna moves in a clockwise manner and the helix in the descending strand contracts in a manner akin to the relaxation of an extended spring. A number of spontaneous mutations in this antenna region cause the hyperinsulinism/hyperammonemia syndrome by decreasing GDH sensitivity to the inhibitor, GTP. Since these residues do not directly contact the bound GTP, the conformational changes in the antenna are apparently crucial to GTP inhibition. In the open conformation, the GTP binding site is distorted such that it can no longer bind GTP. In contrast, ADP binding benefits by the opening of the catalytic cleft since R463 on the pivot helix is pushed into contact distance with the beta-phosphate of ADP. These results support the previous proposal that purines regulate GDH activity by altering the dynamics of the NAD binding domain. Finally, a possible structural mechanism for negative cooperativity is presented.

  16. The role of NAD(+)-dependent isocitrate dehydrogenase 3 subunit α in AFB1 induced liver lesion.

    Science.gov (United States)

    Yang, Chi; Fan, Jue; Zhuang, Zhenhong; Fang, Yi; Zhang, Yanfeng; Wang, Shihua

    2014-01-30

    Aflatoxin B1 (AFB1) is a potent hepatocarcinogen that causes carcinogenesis in many animal species. In previous study, we found that isocitrate dehydrogenasesubunit (IDH3α) was upregulated in AFB1-induced carcinogenesis process. In this study, the sequences of IDH3α from various species were compared and the protein expression levels in different organs were examined, and the results showed that IDH3α was a widely distributed protein and shared highly conserved sequence in various species. In the same time, IDH3α was demonstrated to accumulate in a dose-dependent manner induced by AFB1 in cells, and was also up-regulated in the process of AFB1-induced liver lesion. Similar results were observed when H2O2 was used to replace AFB1. Over-expression of IDH3α increased the phosphorylation level of Akt (Protein kinase B) and neutralized the cellular toxicity induced by AFB1 or H2O2 and apoptosis induced by AFB1, while the reduced expression of IDH3α by siRNA decreased the phosphorylation, indicating that IDH3α played important roles in oxidative stress-induced PI3K/Akt pathway. Overall, the results suggested that AFB1 treatment could increase the expression of IDH3α, and the activated PI3K/Akt pathway by IDH3α eventually neutralized the apoptosis induced by AFB1.

  17. A split and rearranged nuclear gene encoding the iron-sulfur subunit of mitochondrial succinate dehydrogenase in Euglenozoa

    Directory of Open Access Journals (Sweden)

    Gray Michael W

    2009-02-01

    Full Text Available Abstract Background Analyses based on phylogenetic and ultrastructural data have suggested that euglenids (such as Euglena gracilis, trypanosomatids and diplonemids are members of a monophyletic lineage termed Euglenozoa. However, many uncertainties are associated with phylogenetic reconstructions for ancient and rapidly evolving groups; thus, rare genomic characters become increasingly important in reinforcing inferred phylogenetic relationships. Findings We discovered that the iron-sulfur subunit (SdhB of mitochondrial succinate dehydrogenase is encoded by a split and rearranged nuclear gene in Euglena gracilis and trypanosomatids, an example of a rare genomic character. The two subgenic modules are transcribed independently and the resulting mRNAs appear to be independently translated, with the two protein products imported into mitochondria, based on the presence of predicted mitochondrial targeting peptides. Although the inferred protein sequences are in general very divergent from those of other organisms, all of the required iron-sulfur cluster-coordinating residues are present. Moreover, the discontinuity in the euglenozoan SdhB sequence occurs between the two domains of a typical, covalently continuous SdhB, consistent with the inference that the euglenozoan 'half' proteins are functional. Conclusion The discovery of this unique molecular marker provides evidence for the monophyly of Euglenozoa that is independent of evolutionary models. Our results pose questions about the origin and timing of this novel gene arrangement and the structure and function of euglenozoan SdhB.

  18. Mutations in Succinate Dehydrogenase Subunit C Increase Radiosensitivity and Bystander Responses

    Science.gov (United States)

    Zhou, Hongning; Hei, Tom K.

    Although radiation-induced bystander effect is well studied in the past decade, the precise mech-anisms are still unclear. It is likely that a combination of pathways involving both primary and secondary signaling processes is involved in producing a bystander effect. There is recent evidence that mitochondria play a critical role in bystander responses. Recently studies found that a mutation in succinate dehydrogenese subunit C (SDHC), an integral membrane protein in complex II of the electron transport chain, resulted in increased superoxide, oxidative stress, apoptosis, tumorigenesis, and genomic instability, indicating that SDHC play a critical role in maintaining mitochondrial function. In the present study, using Chinese hamster fibroblasts (B1 cells) and the mutants (B9 cells) containing a single base substitution that produced a premature stop codon resulting in a 33-amino acid COOH-terminal truncation of the SDHC protein, we found that B9 cells had an increase in intracellular superoxide content, nitric oxide species, and mitochondrial membrane potential when compared with wild type cells. After irradiated with a grade of doses of gamma rays, B9 cells show an increased radiosensitivity, especially at high doses. The HPRT- mutant yield after gamma-ray irradiation in B9 cells was significantly higher than that of B1 cells. A single, 3Gy dose of gamma-rays increased the background mutant level by more than 4 fold. In contrast, the mutant induction was less than 2 fold in B1 cells. In addition, B9 cells produced a higher bystander mutagenesis after alpha particle irradiation than the B1 cells. Furthermore, pretreated with carboxy-2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO), a nitric oxide scavenger, significantly decreased the bystander effect. Our findings demonstrate that a mutation in SDHC increases radiosensitivity in both directly irradiated cells and in neighboring bystander cells, and mito-chondrial function play an essential role in

  19. The single NqrB and NqrC subunits in the Na(+)-translocating NADH: quinone oxidoreductase (Na(+)-NQR) from Vibrio cholerae each carry one covalently attached FMN.

    Science.gov (United States)

    Casutt, Marco S; Schlosser, Andreas; Buckel, Wolfgang; Steuber, Julia

    2012-10-01

    The Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR) is the prototype of a novel class of flavoproteins carrying a riboflavin phosphate bound to serine or threonine by a phosphodiester bond to the ribityl side chain. This membrane-bound, respiratory complex also contains one non-covalently bound FAD, one non-covalently bound riboflavin, ubiquinone-8 and a [2Fe-2S] cluster. Here, we report the quantitative analysis of the full set of flavin cofactors in the Na(+)-NQR and characterize the mode of linkage of the riboflavin phosphate to the membrane-bound NqrB and NqrC subunits. Release of the flavin by β-elimination and analysis of the cofactor demonstrates that the phosphate group is attached at the 5'-position of the ribityl as in authentic FMN and that the Na(+)-NQR contains approximately 1.7mol covalently bound FMN per mol non-covalently bound FAD. Therefore, each of the single NqrB and NqrC subunits in the Na(+)-NQR carries a single FMN. Elimination of the phosphodiester bond yields a dehydro-2-aminobutyrate residue, which is modified with β-mercaptoethanol by Michael addition. Proteolytic digestion followed by mass determination of peptide fragments reveals exclusive modification of threonine residues, which carry FMN in the native enzyme. The described reactions allow quantification and localization of the covalently attached FMNs in the Na(+)-NQR and in related proteins belonging to the Rhodobacter nitrogen fixation (RNF) family of enzymes. This article is part of a Special Issue entitled: 17th European Bioenergetics Conference (EBEC 2012).

  20. Ovarian expression of inhibin-subunits, 3β-hydroxysteroid dehydrogenase, and cytochrome P450 aromatase during the estrous cycle and pregnancy of shiba goats (Capra hircus).

    Science.gov (United States)

    Kandiel, Mohamed M M; Watanabe, Gen; Taya, Kazuyoshi

    2010-01-01

    The cellular localization of the inhibin subunits (α, β(A), and β (B)), steroidogenic enzymes (3β-hydroxysteroid dehydrogenase (3βHSD) and cytochrome P450 aromatase (P450arom) were evaluated in the ovaries of cyclic (n=6) and pregnant (n=2) Shiba goats (Capra Hircus). The immunointensity of inhibin α and β(A) subunits showed an increase in the granulosa cells (GC) of developing follicles. Inhibin β(B) subunit and P450arom showed high expression in GC of antral follicles. 3βHSD immunoreactivity was uniform in preantral and antral follicles. In follicular phase and late pregnancy, there was a strong expression of inhibin α subunit in GC of antral follicles. Although in mid pregnancy, antral follicles GC showed moderate immunostaining of inhibin β subunits, the immunoreactivity of inhibin β(A) and β(B) subunits was high during the follicular and luteal stages, respectively. While, immunoreactivity of GC to P450arom was moderate during all studied stages, and 3βHSD immunoreactivity was plentiful in antral follicles during the luteal phase. The immunoreactivity to inhibin α subunit and P450arom was abundant during mid pregnancy in the luteal tissues. Immunoreaction to inhibin β subunits was faint-to-moderate in cyclic and pregnancy corpora lutea. Immunoexpression of 3βHSD was maximal in late pregnancy corpora lutea. The present results suggest that, in goats, the GC of antral follicles are the main source of dimeric inhibins and that corpora lutea may partially participate in the secretion of inhibin. Changes in ovarian hormonal levels might depend on the synthesizing capacity of hormones in the follicles and corpora lutea to regulate the goat's reproductive stages.

  1. The β and γ subunits play distinct functional roles in the α2βγ heterotetramer of human NAD-dependent isocitrate dehydrogenase

    Science.gov (United States)

    Ma, Tengfei; Peng, Yingjie; Huang, Wei; Liu, Yabing; Ding, Jianping

    2017-01-01

    Human NAD-dependent isocitrate dehydrogenase existing as the α2βγ heterotetramer, catalyzes the decarboxylation of isocitrate into α-ketoglutarate in the Krebs cycle, and is allosterically regulated by citrate, ADP and ATP. To explore the functional roles of the regulatory β and γ subunits, we systematically characterized the enzymatic properties of the holoenzyme and the composing αβ and αγ heterodimers in the absence and presence of regulators. The biochemical and mutagenesis data show that αβ and αγ alone have considerable basal activity but the full activity of α2βγ requires the assembly and cooperative function of both heterodimers. α2βγ and αγ can be activated by citrate or/and ADP, whereas αβ cannot. The binding of citrate or/and ADP decreases the S0.5,isocitrate and thus enhances the catalytic efficiencies of the enzymes, and the two activators can act independently or synergistically. Moreover, ATP can activate α2βγ and αγ at low concentration and inhibit the enzymes at high concentration, but has only inhibitory effect on αβ. Furthermore, the allosteric activation of α2βγ is through the γ subunit not the β subunit. These results demonstrate that the γ subunit plays regulatory role to activate the holoenzyme, and the β subunit the structural role to facilitate the assembly of the holoenzyme.

  2. Increased sensitivity of photosynthesis to antimycin A induced by inactivation of the chloroplast ndhB gene. Evidence for a participation of the NADH-dehydrogenase complex to cyclic electron flow around photosystem I.

    Science.gov (United States)

    Joët, T; Cournac, L; Horvath, E M; Medgyesy, P; Peltier, G

    2001-04-01

    Tobacco (Nicotiana tabacum var Petit Havana) ndhB-inactivated mutants (ndhB-) obtained by plastid transformation (E.M. Horvath, S.O. Peter, T. Joët, D. Rumeau, L. Cournac, G.V. Horvath, T.A. Kavanagh, C. Schäfer, G. Peltier, P. MedgyesyHorvath [2000] Plant Physiol 123: 1337-1350) were used to study the role of the NADH-dehydrogenase complex (NDH) during photosynthesis and particularly the involvement of this complex in cyclic electron flow around photosystem I (PSI). Photosynthetic activity was determined on leaf discs by measuring CO2 exchange and chlorophyll fluorescence quenchings during a dark-to-light transition. In the absence of treatment, both non-photochemical and photochemical fluorescence quenchings were similar in ndhB- and wild type (WT). When leaf discs were treated with 5 microM antimycin A, an inhibitor of cyclic electron flow around PSI, both quenchings were strongly affected. At steady state, maximum photosynthetic electron transport activity was inhibited by 20% in WT and by 50% in ndhB-. Under non-photorespiratory conditions (2% O2, 2,500 microL x L(-1) CO2), antimycin A had no effect on photosynthetic activity of WT, whereas a 30% inhibition was observed both on quantum yield of photosynthesis assayed by chlorophyll fluorescence and on CO2 assimilation in ndhB-. The effect of antimycin A on ndhB- could not be mimicked by myxothiazol, an inhibitor of the mitochondrial cytochrome bc1 complex, therefore showing that it is not related to an inhibition of the mitochondrial electron transport chain but rather to an inhibition of cyclic electron flow around PSI. We conclude to the existence of two different pathways of cyclic electron flow operating around PSI in higher plant chloroplasts. One of these pathways, sensitive to antimycin A, probably involves ferredoxin plastoquinone reductase, whereas the other involves the NDH complex. The absence of visible phenotype in ndhB- plants under normal conditions is explained by the complement of these two

  3. The human mitochondrial NADH: Ubiquinone oxidoreductase 51-kDa subunit oxidoreductase 51-kDa subunit maps adjacent to the glutathione S-transferase P1-1 gene on chromosome 11q13

    Energy Technology Data Exchange (ETDEWEB)

    Spencer, S.R.; Taylor, J.B.; Cowell, I.G.; Xia, C.L.; Pemble, S.E.; Ketterer, B. (Univ. College and Middlesex School of Medicine, London (United Kingdom))

    1992-12-01

    The soluble glutathione transferases (GSTs) are a family of dimeric isoenymes catalyzing the conjugation of glutathione to hydrophobic electropiles. Their subunits can be grouped into four families, alpha, mu, pi, and theta, on the basis of their primary structures. In man, the pi class is represented by a single gene, GSTP1-1 (GST[pi]) localized to human chromosome 11, band q13. The oncogenes INT2, HSTF1, and PRAD1 are also localized at 11q13, and together with the GSTP1 locus and other gene loci mapped to 11q13, i.e., BCL1 and EMS1, they form a unit of DNA approximately 2000-2500 kb, known as the 11q13 amplicon, which is often amplified in a range of solid tumors. Any gene locus at 11q13 is of interest because it may influence tumorigenesis. 14 refs., 1 fig.

  4. Detection of the gene encoding the small subunit of the CO dehydrogenase enzyme in the H{sub 2}-evolving bacterium Rubrivivax gelatinosus CBS

    Energy Technology Data Exchange (ETDEWEB)

    Kish, A.; Levin, D. [Victoria Univ., BC (Canada)]|[Victoria Univ., BC (Canada)

    2001-06-01

    A purple non-sulfur bacterium, Rubrivivax gelatinosus CBS presents great opportunities, on a commercial scale, for the biological hydrogen production. A water-gas shift reaction is catalyzed when the bacterium is cultured in the presence of carbon oxide in the dark. The result is carbon monoxide (and water) being shifted into hydrogen (H{sub 2}) and carbon dioxide in near stoichiometric quantities. The production of hydrogen as a clean alternative fuel could be accomplished by using carbon monoxide generated from gasified waste biomass, using the bacterial water-gas shift reaction for that purpose. The characterization of three key enzymes and the genes encoding them was performed in a closely related purple non-sulfur bacterium called Rhodospirillum rubrum. They were: (1) a carbon monoxide dehydrogenase (CODH), (2) the ferredoxin-like electron-carrier small subunit of the CODH enzyme, and (3) an hydrogen-evolving hydrogenase. A transcriptional unit separate from the genes encoding the CODH and its ferredoxin-like small subunit encode the genes for the hydrogenase. A fragment of the Rhodospirillum rubrum ferredoxin-like subunit gene was amplified through the use of a polymerase chain reaction. Southern blots of restriction endonuclease digested genomic deoxyribonucleic acid (DNA) extracted from Rubrivivax gelatinosus CBS was probed with the fragment of the Rhodospirillum rubrum previously amplified using the polymerase chain reaction. Confirmation of the identification is being confirmed, while the gene is sequenced. 25 refs., 2 figs.

  5. Characterization of Autoantibodies against the E1 Subunit of Branched-Chain 2-Oxoacid Dehydrogenase in Patients with Primary Biliary Cirrhosis

    Directory of Open Access Journals (Sweden)

    Tsutomu Mori

    2012-01-01

    Full Text Available Primary biliary cirrhosis (PBC is characterized by antimitochondrial antibodies (AMAs that react with the lipoyl-containing E2 subunits of 2-oxoacid dehydrogenase complexes such as BCOADC and PDC. The lipoyl domains of E2 contain the major epitopes essential for immunopathology. However, the non-lipoyl-containing E1 subunits are also frequently targeted. Since anti-E1 antibodies always appear in combination with anti-E2 antibodies, the mechanisms underlying the autoimmunity against E1 may be linked to, but distinct from, those against E2. Here, we demonstrate that intermolecular and intramolecular determinant spreading underlies the autoimmunity against E1. We performed characterizations and epitope mapping for anti-BCOADC-E1 antibodies from both the intermolecular and intramolecular points of view. The antibody reactivities form a cluster against the BCOADC complex that is distinct from that against the PDC complex, and the anti-BCOADC-E1 antibodies arise as part of the cluster against the BCOADC complex. Multiple epitopes are present on the surface of the BCOADC-E1 molecule, and the major epitope overlaps with the active center. Sera with anti-BCOADC-E1 antibodies strongly inhibited the enzyme activity. These findings suggest that the E1 subunit as part of the native BCOADC complex is an immunogen, and that determinant spreading is involved in the pathogenesis of AMA production.

  6. Functional Characterization of the Subunits N, H, J, and O of the NAD(P)H Dehydrogenase Complexes in Synechocystis sp. Strain PCC 6803.

    Science.gov (United States)

    He, Zhihui; Mi, Hualing

    2016-06-01

    The cyanobacterial NAD(P)H dehydrogenase (NDH-1) complexes play crucial roles in variety of bioenergetic reactions such as respiration, CO2 uptake, and cyclic electron transport around PSI. Recently, substantial progress has been made in identifying the composition of subunits of NDH-1 complexes. However, the localization and the physiological roles of several subunits in cyanobacteria are not fully understood. Here, by constructing fully segregated ndhN, ndhO, ndhH, and ndhJ null mutants in Synechocystis sp. strain PCC 6803, we found that deletion of ndhN, ndhH, or ndhJ but not ndhO severely impaired the accumulation of the hydrophilic subunits of the NDH-1 in the thylakoid membrane, resulting in disassembly of NDH-1MS, NDH-1MS', as well as NDH-1L, finally causing the severe growth suppression phenotype. In contrast, deletion of NdhO affected the growth at pH 6.5 in air. In the cytoplasm, either NdhH or NdhJ deleted mutant, but neither NdhN nor NdhO deleted mutant, failed to accumulate the NDH-1 assembly intermediate consisting of NdhH, NdhJ, NdhK, and NdhM. Based on these results, we suggest that NdhN, NdhH, and NdhJ are essential for the stability and the activities of NDH-1 complexes, while NdhO for NDH-1 functions under the condition of inorganic carbon limitation in Synechocystis sp. strain PCC 6803. We discuss the roles of these subunits and propose a new NDH-1 model. © 2016 American Society of Plant Biologists. All Rights Reserved.

  7. Novel O-palmitolylated beta-E1 subunit of pyruvate dehydrogenase is phosphorylated during ischemia/reperfusion injury

    Directory of Open Access Journals (Sweden)

    Barr Amy J

    2010-07-01

    Full Text Available Abstract Background During and following myocardial ischemia, glucose oxidation rates are low and fatty acids dominate as a source of oxidative metabolism. This metabolic phenotype is associated with contractile dysfunction during reperfusion. To determine the mechanism of this reliance on fatty acid oxidation as a source of ATP generation, a functional proteomics approach was utilized. Results 2-D gel electrophoresis of mitochondria from working rat hearts subjected to 25 minutes of global no flow ischemia followed by 40 minutes of aerobic reperfusion identified 32 changes in protein abundance compared to aerobic controls. Of the five proteins with the greatest change in abundance, two were increased (long chain acyl-coenzyme A dehydrogenase (48 ± 1 versus 39 ± 3 arbitrary units, n = 3, P In silico analysis identified the putative kinases as the insulin receptor kinase for the more basic form and protein kinase Cζ or protein kinase A for the more acidic form. These modifications of pyruvate dehydrogenase are associated with a 35% decrease in glucose oxidation during reperfusion. Conclusions Cardiac ischemia/reperfusion induces significant changes to a number of metabolic proteins of the mitochondrial proteome. In particular, ischemia/reperfusion induced the post-translational modification of pyruvate dehydrogenase, the rate-limiting step of glucose oxidation, which is associated with a 35% decrease in glucose oxidation during reperfusion. Therefore these post-translational modifications may have important implications in the regulation of myocardial energy metabolism.

  8. Gene transcript accumulation and in situ mRNA hybridization of two putative glutamate dehydrogenase genes in etiolated Glycine max seedlings.

    Science.gov (United States)

    Dimou, M; Tsaniklidis, G; Aivalakis, G; Katinakis, P

    2015-01-01

    Glutamate dehydrogenase (EC 1.4.1.2) is a multimeric enzyme that catalyzes the reversible amination of α-ketoglutarate to form glutamate. We characterized cDNA clones of two Glycine max sequences, GmGDH1 and GmGDH2, that code for putative α- and β-subunits, respectively, of the NADH dependent enzyme. Temporal and spatial gene transcript accumulation studies using semiquantitative RT-PCR and in situ hybridization have shown an overlapping gene transcript accumulation pattern with differences in relative gene transcript accumulation in the organs examined. Detection of NADH-dependent glutamate dehydrogenase activity in situ using a histochemical method showed concordance with the spatial gene transcript accumulation patterns. Our findings suggest that although the two gene transcripts are co-localized in roots of etiolated soybean seedlings, the ratio of the two subunits of the active holoenzyme may vary among tissues.

  9. Role of positron emission tomography and bone scintigraphy in the evaluation of bone involvement in metastatic pheochromocytoma and paraganglioma: specific implications for succinate dehydrogenase enzyme subunit B gene mutations.

    NARCIS (Netherlands)

    Zelinka, T.; Timmers, H.J.L.M.; Kozupa, A.; Chen, C.C.; Carrasquillo, J.A.; Reynolds, J.C.; Ling, A.; Eisenhofer, G.; Lazurova, I.; Adams, K.T.; Whatley, M.A.; Widimsky, J.Jr.; Pacak, K.

    2008-01-01

    We performed a retrospective analysis of 71 subjects with metastatic pheochromocytoma and paraganglioma (30 subjects with mutation of succinate dehydrogenase enzyme subunit B (SDHB) gene and 41 subjects without SDHB mutation). Sixty-nine percent presented with bone metastases (SDHB +/-: 77% vs 63%),

  10. Molecular characterization of Fasciola hepatica and phylogenetic analysis based on mitochondrial (nicotiamide adenine dinucleotide dehydrogenase subunit I and cytochrome oxidase subunit I) genes from the North-East of Iran

    Science.gov (United States)

    Reaghi, Saber; Haghighi, Ali; Harandi, Majid Fasihi; Spotin, Adel; Arzamani, Kourosh; Rouhani, Soheila

    2016-01-01

    Aim: Fascioliasis is one of the most zoonotic diseases with global extension. As the epidemiological distribution of Fasciola may lead to various genetic patterns of the parasite, the aim of this study is to identify Fasciola hepatica based on spermatogenesis, and phylogenetic analysis using mitochondrial (nicotiamide adenine dinucleotide dehydrogenase subunit I [ND1] and cytochrome oxidase subunit I) gene marker. Materials and Methods: In this study, 90 F. hepatica collected from 30 cattle at slaughterhouse located in three different geographical locations in the North-East of Iran were evaluated based on spermatogenetic ability and internal transcribed spacer 1 gene restriction fragment length polymorphism pattern. Genetic diversity and phylogenetic relationship using mtDNA gene marker for the isolates from the North-East of Iran, and other countries were then analyzed. Results: Partial sequences of mtDNA showed eight haplotypes in both genes. The phylogenic analysis using neighbor joining as well as maximum likelihood methods showed similar topologies of trees. Pairwise fixation index between different F. hepatica populations calculated from the nucleotide data set of ND1 gene are statistically significant and show the genetic difference. Conclusion: F. hepatica found in this region of Iran has different genetic structures through the other Fasciola populations in the world. PMID:27733809

  11. Automated High Throughput Protein Crystallization Screening at Nanoliter Scale and Protein Structural Study on Lactate Dehydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Li, Fenglei [Iowa State Univ., Ames, IA (United States)

    2006-08-09

    , evaporation rate can be controlled or adjusted in this method during the crystallization process to favor either nucleation or growing processes for optimizing crystallization process. The protein crystals gotten by this method were experimentally proven to possess high x-ray diffraction qualities. Finally, we crystallized human lactate dehydrogenase 1 (H4) complexed with NADH and determined its structure by x-ray crystallography. The structure of LDH/NADH displays a significantly different structural feature, compared with LDH/NADH/inhibitor ternary complex structure, that subunits in LDH/NADH complex show open conformation or two conformations on the active site while the subunits in LDH/NADH/inhibitor are all in close conformation. Multiple LDH/NADH crystals were obtained and used for x-ray diffraction experiments. Difference in subunit conformation was observed among the structures independently solved from multiple individual LDH/NADH crystals. Structural differences observed among crystals suggest the existence of multiple conformers in solution.

  12. NdhO, a subunit of NADPH dehydrogenase, destabilizes medium size complex of the enzyme in Synechocystis sp. strain PCC 6803.

    Science.gov (United States)

    Zhao, Jiaohong; Gao, Fudan; Zhang, Jingsong; Ogawa, Teruo; Ma, Weimin

    2014-09-26

    Two mutants that grew faster than the wild-type (WT) strain under high light conditions were isolated from Synechocystis sp. strain PCC 6803 transformed with a transposon-bearing library. Both mutants had a tag in ssl1690 encoding NdhO. Deletion of ndhO increased the activity of NADPH dehydrogenase (NDH-1)-dependent cyclic electron transport around photosystem I (NDH-CET), while overexpression decreased the activity. Although deletion and overexpression of ndhO did not have significant effects on the amount of other subunits such as NdhH, NdhI, NdhK, and NdhM in the cells, the amount of these subunits in the medium size NDH-1 (NDH-1M) complex was higher in the ndhO-deletion mutant and much lower in the overexpression strain than in the WT. NdhO strongly interacts with NdhI and NdhK but not with other subunits. NdhI interacts with NdhK and the interaction was blocked by NdhO. The blocking may destabilize the NDH-1M complex and repress the NDH-CET activity. When cells were transferred from growth light to high light, the amounts of NdhI and NdhK increased without significant change in the amount of NdhO, thus decreasing the relative amount of NdhO. This might have decreased the blocking, thereby stabilizing the NDH-1M complex and increasing the NDH-CET activity under high light conditions. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. NCBI nr-aa BLAST: CBRC-PCAP-01-1704 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available rogenase subunit 1 [Formica rufa] gb|AAR86956.1| NADH dehydrogenase subunit 1 [Formica rufa...nsis] gb|AAR86998.1| NADH dehydrogenase subunit 1 [Formica pratensis] gb|AAR91932.1| NADH dehydrogenase subunit 1 [Formica rufa...] gb|AAR91934.1| NADH dehydrogenase subunit 1 [Formica rufa] gb|AAR91936.1| NADH dehydrog...enase subunit 1 [Formica rufa] gb|AAR91938.1| NADH dehydrogenase subunit 1 [Formi

  14. EST Table: FS878523 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18196.1| NADH dehydrogenase subunit 2 [Bombyx mandarina] gb|ADE18209.1| ...NADH dehydrogenase subunit 2 [Bombyx mandarina] gb|ADE18274.1| NADH dehydrogenase subunit 2 [Bombyx mandarin...a] gb|ADE18287.1| NADH dehydrogenase subunit 2 [Bombyx mandarina] gb|ADE18391.1| ...NADH dehydrogenase subunit 2 [Bombyx mandarina] gb|ADE18521.1| NADH dehydrogenase subunit 2 [Bombyx mandarin...a] gb|ADE18560.1| NADH dehydrogenase subunit 2 [Bombyx mandarina] gb|ADE18586.1| NADH dehydrogenase subunit 2 [Bombyx mandarin

  15. Identification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.

    Directory of Open Access Journals (Sweden)

    Zhenhong Sun

    Full Text Available The pathogen Mycoplasma bovis (M. bovis is a major cause of respiratory disease, mastitis, and arthritis in cattle. Screening the key immunogenic proteins and updating rapid diagnostic techniques are necessary to the prevention and control of M. bovis infection. In this study, 19 highly immunogenic proteins from M. bovis strain PD were identified using 2-dimensional gel electrophoresis, immunoblotting and MALDI-TOF/TOF MS. Of these 19 proteins, pyruvate dehydrogenase E1 component beta subunit (PDHB showed excellent immune reactivity and repeatability. PDHB was found to be conserved in different M. bovis isolates, as indicated by Western blot analysis. On the basis of these results, a rPDHB-based indirect ELISA (iELISA was established for the detection of serum antibodies using prokaryotically expressed recombinant PDHB protein as the coating antigen. The specificity analysis result showed that rPDHB-based iELISA did not react with other pathogens assessed in our study except M. agalactiae (which infects sheep and goats. Moreover, 358 serum samples from several disease-affected cattle feedlots were tested using this iELISA system and a commercial kit, which gave positive rates of 50.8% and 39.9%, respectively. The estimated Kappa agreement coefficient between the two methods was 0.783. Notably, 39 positive serum samples that had been missed by the commercial kit were all found to be positive by Western blot analysis. The detection rate of rPDHB-based iELISA was significantly higher than that of the commercial kit at a serum dilution ratio of 1∶5120 to 1∶10,240 (P<0.05. Taken together, these results provide important information regarding the novel immunogenic proteins of M. bovis. The established rPDHB-based iELISA may be suitable for use as a new method of antibody detection in M. bovis.

  16. Succinate Dehydrogenase Subunit B (SDHB Is Expressed in Neurofibromatosis 1-Associated Gastrointestinal Stromal Tumors (Gists: Implications for the SDHB Expression Based Classification of Gists

    Directory of Open Access Journals (Sweden)

    Jeanny H. Wang, Jerzy Lasota, Markku Miettinen

    2011-01-01

    Full Text Available Gastrointestinal Stromal Tumor (GIST is the most common mesenchymal tumor of the digestive tract. GISTs develop with relatively high incidence in patients with Neurofibromatosis-1 syndrome (NF1. Mutational activation of KIT or PDGFRA is believed to be a driving force in the pathogenesis of familial and sporadic GISTs. Unlike those tumors, NF1-associated GISTs do not have KIT or PGDFRA mutations. Similarly, no mutational activation of KIT or PDGFRA has been identified in pediatric GISTs and in GISTs associated with Carney Triad and Carney-Stratakis Syndrome. KIT and PDGFRA-wild type tumors are expected to have lesser response to imatinib treatment. Recently, Carney Triad and Carney-Stratakis Syndrome -associated GISTs and pediatric GISTs have been shown to have a loss of expression of succinate dehydrogenase subunit B (SDHB, a Krebs cycle/electron transport chain interface protein. It was proposed that GISTs can be divided into SDHB- positive (type 1, and SDHB-negative (type 2 tumors because of similarities in clinical features and response to imatinib treatment. In this study, SDHB expression was examined immunohistochemically in 22 well-characterized NF1-associated GISTs. All analyzed tumors expressed SDHB. Based on SDHB-expression status, NF1-associated GISTs belong to type 1 category; however, similarly to SDHB type 2 tumors, they do not respond well to imatinib treatment. Therefore, a simple categorization of GISTs into SDHB-positive and-negative seems to be incomplete. A classification based on both SDHB expression status and KIT and PDGFRA mutation status characterize GISTs more accurately and allow subdivision of SDHB-positive tumors into different clinico-genetic categories.

  17. Construction of NADH Regeneration System in Klebisella pneumoniae with Aldehyde Dehydrogenase Inactivated%在Klebsiella Pneumoniae醛脱氢酶失活菌中构建NADH再生系统

    Institute of Scientific and Technical Information of China (English)

    黄志华; 张延平; 黄星; 王宝光; 曹竹安

    2006-01-01

    生物法生产1,3-丙二醇(1,3-Propanediol,1,3-PD)是当前工业生物技术研究的热点之一,生产过程中,需要消耗还原当量NADH,NADH的有效供给决定了1,3-PD的产量和得率.采用PCR的方法从Candida boidinii基因组中克隆编码fdh的基因,将该基因片段插入载体pMALTM-p2X,构建表达载体pMALTM-p2X-fdh,并转入醛脱氢酶失活菌Klebsiella pneumoniae DA-1HB,获得重组菌Klebsiella pneumoniae DAF-1.在IPTG浓度0.5 mmol/L时,诱导3 h后甲酸脱氢酶表达明显;发酵过程中甲酸脱氢酶比酶活达到4.82 U/mg;与出发菌株K.pneumoniae DA-1HB相比,重组菌DAF-1合成1,3-丙二醇的浓度提高了19.2%.

  18. Control of oxygen free radical formation from mitochondrial complex I: roles for protein kinase A and pyruvate dehydrogenase kinase.

    Science.gov (United States)

    Raha, Sandeep; Myint, A Tomoko; Johnstone, Leslie; Robinson, Brian H

    2002-03-01

    Human NADH CoQ oxidoreductase is composed of a total of 43 subunits and has been demonstrated to be a major site for the production of superoxide by mitochondria. Incubation of rat heart mitochondria with ATP resulted in the phosphorylation of two mitochondrial membrane proteins, one with a M(r) of 6 kDa consistent with the NDUFA1 (MWFE), and one at 18kDa consistent with either NDUFS4 (AQDQ) or NDUFB7 (B18). Phosphorylation of both subunits was enhanced by cAMP derivatives and protein kinase A (PKA) and was inhibited by PKA inhibitors (PKAi). When mitochondrial membranes were incubated with pyruvate dehydrogenase kinase, phosphorylation of an 18kDa protein but not a 6kDa protein was observed. NADH cytochrome c reductase activity was decreased and superoxide production rates with NADH as substrate were increased. On the other hand, with protein kinase A-driven phosphorylation, NADH cytochrome c reductase was increased and superoxide production decreased. Overall there was a 4-fold variation in electron transport rates observable at the extremes of these phosphorylation events. This suggests that electron flow through complex I and the production of oxygen free radicals can be regulated by phosphorylation events. In light of these observations we discuss a potential model for the dual regulation of complex I and the production of oxygen free radicals by both PKA and PDH kinase.

  19. Biochemical characterization of a recombinant short-chain NAD(H)-dependent dehydrogenase/reductase from Sulfolobus acidocaldarius.

    Science.gov (United States)

    Pennacchio, Angela; Giordano, Assunta; Pucci, Biagio; Rossi, Mosè; Raia, Carlo A

    2010-03-01

    The gene encoding a novel alcohol dehydrogenase that belongs to the short-chain dehydrogenases/reductases (SDRs) superfamily was identified in the aerobic thermoacidophilic crenarchaeon Sulfolobus acidocaldarius strain DSM 639. The saadh gene was heterologously overexpressed in Escherichia coli, and the protein (SaADH) was purified to homogeneity and characterized. SaADH is a tetrameric enzyme consisting of identical 28,978-Da subunits, each composed of 264 amino acids. The enzyme has remarkable thermophilicity and thermal stability, displaying activity at temperatures up to 75 degrees C and a 30-min half-inactivation temperature of ~90 degrees C, and shows good tolerance to common organic solvents. SaADH has a strict requirement for NAD(H) as the coenzyme, and displays a preference for the reduction of alicyclic, bicyclic and aromatic ketones and alpha-keto esters, but is poorly active on aliphatic, cyclic and aromatic alcohols, and shows no activity on aldehydes. The enzyme catalyses the reduction of alpha-methyl and alpha-ethyl benzoylformate, and methyl o-chlorobenzoylformate with 100% conversion to methyl (S)-mandelate [17% enantiomeric excess (ee)], ethyl (R)-mandelate (50% ee), and methyl (R)-o-chloromandelate (72% ee), respectively, with an efficient in situ NADH-recycling system which involves glucose and a thermophilic glucose dehydrogenase. This study provides further evidence supporting the critical role of the D37 residue in discriminating NAD(H) from NAD(P)H in members of the SDR superfamily.

  20. Thermal stabilization of formaldehyde dehydrogenase by encapsulation in liposomes with nicotinamide adenine dinucleotide.

    Science.gov (United States)

    Yoshimoto, Makoto; Yamashita, Takayuki; Kinoshita, Satoshi

    2011-07-10

    The thermal stability of formaldehyde dehydrogenase (FaDH) from Pseudomonas sp. was examined and controlled by encapsulation in liposomes with β-reduced nicotinamide adenine dinucleotide (NADH). The activity of 4.8 μg/mL free FaDH at pH 8.5 in catalyzing the oxidation of 50mM formaldehyde was highly dependent on temperature so that the activity at 60 °C was 27 times larger than that at 25 °C. Thermal stability of the FaDH activity was examined with and without liposomes composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). Rapid deactivation of free FaDH was observed at 60 °C because of its dissociation into two subunits. The rate of dissociative deactivation of POPC liposome-encapsulated FaDH was smaller than that of the free enzyme. The liposomal FaDH was however progressively deactivated for the incubation period of 60 min eventually leading to complete loss of its activity. The free FaDH and NADH molecules were revealed to form the thermostable binary complex. The thermal stability of POPC liposome-encapsulated FaDH and NADH system was significantly higher than the liposomal enzyme without cofactor. The above results clearly show that NADH is a key molecule that controls the activity and stability of FaDH in liposomes at high temperatures.

  1. NCBI nr-aa BLAST: CBRC-TTRU-01-1312 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1312 gb|AAC98204.1| NADH dehydrogenase subunit 4 [Haemonchus placei] g...b|AAC98205.1| NADH dehydrogenase subunit 4 [Haemonchus placei] gb|AAC98208.1| NADH dehydrogenase subunit 4 [Haemonchus place...i] gb|AAC98211.1| NADH dehydrogenase subunit 4 [Haemonchus placei] gb|AAC98213.1| NADH dehyd...rogenase subunit 4 [Haemonchus placei] gb|AAC98216.1| NADH dehydrogenase subunit 4 [Haemonchus place...i] gb|AAC98217.1| NADH dehydrogenase subunit 4 [Haemonchus placei] gb|AAC98221.1| NADH de

  2. NdhP is an exclusive subunit of large complex of NADPH dehydrogenase essential to stabilize the complex in Synechocystis sp. strain PCC 6803.

    Science.gov (United States)

    Zhang, Jingsong; Gao, Fudan; Zhao, Jiaohong; Ogawa, Teruo; Wang, Quanxi; Ma, Weimin

    2014-07-04

    Two major complexes of NADPH dehydrogenase (NDH-1) have been identified in cyanobacteria. A large complex (NDH-1L) contains NdhD1 and NdhF1, which are absent in a medium size complex (NDH-1M). They play important roles in respiration, cyclic electron transport around photosystem I, and CO2 acquisition. Two mutants sensitive to high light for growth and impaired in NDH-1-mediated cyclic electron transfer were isolated from Synechocystis sp. strain PCC 6803 transformed with a transposon-bearing library. Both mutants had a tag in sml0013 encoding NdhP, a single transmembrane small subunit of the NDH-1 complex. During prolonged incubation of the wild type thylakoid membrane with n-dodecyl β-d-maltoside (DM), about half of the NDH-1L was disassembled to NDH-1M and the rest decomposed completely without forming NDH-1M. In the ndhP deletion mutant (ΔndhP), disassembling of NDH-1L to NDH-1M occurred even on ice, and decomposition to a small piece occurred at room temperature much faster than in the wild type. Deletion of the C-terminal tail of NdhP gave the same result. The C terminus of NdhP was tagged by YFP-His6. Blue native gel electrophoresis of the DM-treated thylakoid membrane of this strain and Western analysis using the antibody against GFP revealed that NdhP-YFP-His6 was exclusively confined to NDH-1L. During prolonged incubation of the thylakoid membrane of the tagged strain with DM at room temperature, NDH-1L was partially disassembled to NDH-1M and the 160-kDa band containing NdhP-YFP-His6 and possibly NdhD1 and NdhF1. We therefore conclude that NdhP, especially its C-terminal tail, is essential to assemble NdhD1 and NdhF1 and stabilize the NDH-1L complex. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  3. Identification of proteins capable of metal reduction from the proteome of the Gram-positive bacterium Desulfotomaculum reducens MI-1 using an NADH-based activity assay

    Science.gov (United States)

    Otwell, A.E.; Sherwood, R.W.; Zhang, S.; Nelson, O.D.; Li, Z.; Lin, H.; Callister, S.J.; Richardson, R.E.

    2015-01-01

    Summary Understanding of microbial metal reduction is based almost solely on studies of Gram-negative organisms. In this study, we focus on Desulfotomaculum reducens MI-1, a Gram-positive metal reducer whose genome lacks genes with similarity to any characterized metal reductase. Using non-denaturing separations and mass spectrometry identification, in combination with a colorimetric screen for chelated Fe(III)-NTA reduction with NADH as electron donor, we have identified proteins from the D. reducens proteome not previously characterized as iron reductases. Their function was confirmed by heterologous expression in E. coli. Furthermore, we show that these proteins have the capability to reduce soluble Cr(VI) and U(VI) with NADH as electron donor. The proteins identified are NADH:flavin oxidoreductase (Dred_2421) and a protein complex composed of oxidoreductase FAD/NAD(P)-binding subunit (Dred_1685) and dihydroorotate dehydrogenase 1B (Dred_1686). Dred_2421 was identified in the soluble proteome and is predicted to be a cytoplasmic protein. Dred_1685 and Dred_1686 were identified in both the soluble as well as the insoluble protein fraction, suggesting a type of membrane-association, although PSORTb predicts both proteins are cytoplasmic. This study is the first functional proteomic analysis of D. reducens and one of the first analyses of metal and radionuclide reduction in an environmentally relevant Gram-positive bacterium. PMID:25389064

  4. Biochemical and structural characterization of recombinant short-chain NAD(H)-dependent dehydrogenase/reductase from Sulfolobus acidocaldarius highly enantioselective on diaryl diketone benzil.

    Science.gov (United States)

    Pennacchio, Angela; Sannino, Vincenzo; Sorrentino, Giosuè; Rossi, Mosè; Raia, Carlo A; Esposito, Luciana

    2013-05-01

    The gene encoding a novel alcohol dehydrogenase that belongs to the short-chain dehydrogenases/reductases superfamily was identified in the aerobic thermoacidophilic crenarchaeon Sulfolobus acidocaldarius strain DSM 639. The saadh2 gene was heterologously overexpressed in Escherichia coli, and the resulting protein (SaADH2) was purified to homogeneity and both biochemically and structurally characterized. The crystal structure of the SaADH2 NADH-bound form reveals that the enzyme is a tetramer consisting of identical 27,024-Da subunits, each composed of 255 amino acids. The enzyme has remarkable thermophilicity and thermal stability, displaying activity at temperatures up to 80 °C and a 30-min half-inactivation temperature of ∼88 °C. It also shows good tolerance to common organic solvents and a strict requirement for NAD(H) as the coenzyme. SaADH2 displays a preference for the reduction of alicyclic, bicyclic and aromatic ketones and α-ketoesters, but is poorly active on aliphatic, cyclic and aromatic alcohols, showing no activity on aldehydes. Interestingly, the enzyme catalyses the asymmetric reduction of benzil to (R)-benzoin with both excellent conversion (98 %) and optical purity (98 %) by way of an efficient in situ NADH-recycling system involving a second thermophilic ADH. The crystal structure of the binary complex SaADH2-NADH, determined at 1.75 Å resolution, reveals details of the active site providing hints on the structural basis of the enzyme enantioselectivity.

  5. External NAD(P)H dehydrogenases in Acanthamoeba castellanii mitochondria.

    Science.gov (United States)

    Antos-Krzeminska, Nina; Jarmuszkiewicz, Wieslawa

    2014-09-01

    The mitochondrial respiratory chain of plants and some fungi contains multiple rotenone-insensitive NAD(P)H dehydrogenases, of which at least two are located on the outer surface of the inner membrane (i.e., external NADH and external NADPH dehydrogenases). Annotated sequences of the putative alternative NAD(P)H dehydrogenases of the protozoan Acanthamoeba castellanii demonstrated similarity to plant and fungal sequences. We also studied activity of these dehydrogenases in isolated A. castellanii mitochondria. External NADPH oxidation was observed for the first time in protist mitochondria. The coupling parameters were similar for external NADH oxidation and external NADPH oxidation, indicating similar efficiencies of ATP synthesis. Both external NADH oxidation and external NADPH oxidation had an optimal pH of 6.8 independent of relevant ubiquinol-oxidizing pathways, the cytochrome pathway or a GMP-stimulated alternative oxidase. The maximal oxidizing activity with external NADH was almost double that with external NADPH. However, a lower Michaelis constant (K(M)) value for external NADPH oxidation was observed compared to that for external NADH oxidation. Stimulation by Ca(2+) was approximately 10 times higher for external NADPH oxidation, while NADH dehydrogenase(s) appeared to be slightly dependent on Ca(2+). Our results indicate that external NAD(P)H dehydrogenases similar to those in plant and fungal mitochondria function in mitochondria of A. castellanii.

  6. NCBI nr-aa BLAST: CBRC-RNOR-03-0485 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-03-0485 gb|AAY43502.1| NADH dehydrogenase subunit 2 [Milvus milvus milvus...] gb|AAY43505.1| NADH dehydrogenase subunit 2 [Milvus milvus milvus] gb|AAY43507.1| NADH dehydrogenase subunit 2 [Milvus milvu...s milvus] gb|AAY43508.1| NADH dehydrogenase subunit 2 [Milvus milvus fasciicauda] gb|AAY43...509.1| NADH dehydrogenase subunit 2 [Milvus milvus fasciicauda] gb|AAY43511.1| NA...DH dehydrogenase subunit 2 [Milvus milvus fasciicauda] gb|AAY43512.1| NADH dehydrogenase subunit 2 [Milvus milvu

  7. NCBI nr-aa BLAST: CBRC-TTRU-01-1020 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1020 ref|YP_271950.1| NADH dehydrogenase subunit 5 [Montastraea faveol...ata] ref|YP_271937.1| NADH dehydrogenase subunit 5 [Montastraea franksi] ref|YP_271924.1| NADH dehydrogenase subunit 5 [Montastrae...a annularis] dbj|BAE16177.1| NADH dehydrogenase subunit 5 [Montastraea annularis] dbj|...BAE16190.1| NADH dehydrogenase subunit 5 [Montastraea annularis] dbj|BAE16203.1| ...NADH dehydrogenase subunit 5 [Montastraea franksi] dbj|BAE16216.1| NADH dehydrogenase subunit 5 [Montastraea

  8. NCBI nr-aa BLAST: CBRC-DNOV-01-0366 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0366 ref|YP_271950.1| NADH dehydrogenase subunit 5 [Montastraea faveol...ata] ref|YP_271937.1| NADH dehydrogenase subunit 5 [Montastraea franksi] ref|YP_271924.1| NADH dehydrogenase subunit 5 [Montastrae...a annularis] dbj|BAE16177.1| NADH dehydrogenase subunit 5 [Montastraea annularis] dbj|...BAE16190.1| NADH dehydrogenase subunit 5 [Montastraea annularis] dbj|BAE16203.1| ...NADH dehydrogenase subunit 5 [Montastraea franksi] dbj|BAE16216.1| NADH dehydrogenase subunit 5 [Montastraea

  9. NCBI nr-aa BLAST: CBRC-ETEL-01-0265 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ETEL-01-0265 ref|YP_271953.1| NADH dehydrogenase subunit 2 [Montastraea faveol...ata] ref|YP_271940.1| NADH dehydrogenase subunit 2 [Montastraea franksi] ref|YP_271927.1| NADH dehydrogenase subunit 2 [Montastrae...a annularis] dbj|BAE16180.1| NADH dehydrogenase subunit 2 [Montastraea annularis] dbj|...BAE16193.1| NADH dehydrogenase subunit 2 [Montastraea annularis] dbj|BAE16206.1| ...NADH dehydrogenase subunit 2 [Montastraea franksi] dbj|BAE16219.1| NADH dehydrogenase subunit 2 [Montastraea

  10. EST Table: BY916307 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BY916307 mg0096 10/09/28 55 %/142 aa gb|AAP42818.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| NADH dehydrog...enase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE1855...7.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| NADH dehydrog...enase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE1871

  11. EST Table: BB990129 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available one) activity)|GO:0055114(oxidation reduction) 10/09/28 49 %/157 aa gb|AAP42818.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| NADH de...hydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| NADH de...hydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|A

  12. EST Table: AU000518 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available AU000518 e40637 10/09/28 50 %/162 aa gb|AAP42818.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| NADH dehydrog...enase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE1855...7.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| NADH dehydrog...enase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE1871

  13. EST Table: BB983718 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB983718 ovS3024C07r 10/09/28 47 %/160 aa gb|AAP42818.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| NADH deh...ydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|AD...E18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| NADH deh...ydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|AD

  14. EST Table: BB983048 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BB983048 ovS3015B10r 10/09/28 46 %/157 aa gb|AAP42818.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| NADH deh...ydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|AD...E18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| NADH deh...ydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|AD

  15. EST Table: DN985187 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available DN985187 EST01033 10/09/28 51 %/157 aa gb|AAP42818.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| NADH dehydr...ogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18...557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| NADH dehydr...ogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18

  16. EST Table: DN237519 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available DN237519 EST00645 10/09/29 49 %/159 aa gb|AAP42818.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| NADH dehydr...ogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18...557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| NADH dehydr...ogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18

  17. EST Table: BP181108 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BP181108 ovS324C07f 10/09/28 45 %/160 aa gb|AAP42818.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| NADH dehy...drogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE...18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| NADH dehy...drogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE

  18. NCBI nr-aa BLAST: CBRC-MDOM-03-0406 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-03-0406 gb|ABS79776.1| NADH dehydrogenase subunit 5 [Cydia pomonella] gb|...ABS79778.1| NADH dehydrogenase subunit 5 [Cydia pomonella] gb|ABS79780.1| NADH dehydrogenase subunit 5 [Cydia pomonel...la] gb|ABS79781.1| NADH dehydrogenase subunit 5 [Cydia pomonella] gb|ABS79784.1| NADH dehydrogenase subunit 5 [Cydia pomonel...la] gb|ABS79791.1| NADH dehydrogenase subunit 5 [Cydia pomonel...la] gb|ABS79792.1| NADH dehydrogenase subunit 5 [Cydia pomonella] ABS79776.1 0.071 23% ...

  19. NCBI nr-aa BLAST: CBRC-TTRU-01-0408 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0408 gb|AAO16385.1| NADH dehydrogenase subunit 6 [Caenorhabditis elega...ns] gb|AAO16388.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16391.1| NADH dehydrogenase subunit 6 [Caenor...habditis elegans] gb|AAO16394.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AA...O16400.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16406.1| NA...DH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16409.1| NADH dehydrogenase subunit 6 [Caenorhabdi

  20. NCBI nr-aa BLAST: CBRC-TTRU-01-1188 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1188 gb|AAO16385.1| NADH dehydrogenase subunit 6 [Caenorhabditis elega...ns] gb|AAO16388.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16391.1| NADH dehydrogenase subunit 6 [Caenor...habditis elegans] gb|AAO16394.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AA...O16400.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16406.1| NA...DH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16409.1| NADH dehydrogenase subunit 6 [Caenorhabdi

  1. NCBI nr-aa BLAST: CBRC-TTRU-01-0558 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0558 gb|AAO16385.1| NADH dehydrogenase subunit 6 [Caenorhabditis elega...ns] gb|AAO16388.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16391.1| NADH dehydrogenase subunit 6 [Caenor...habditis elegans] gb|AAO16394.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AA...O16400.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16406.1| NA...DH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16409.1| NADH dehydrogenase subunit 6 [Caenorhabdi

  2. NCBI nr-aa BLAST: CBRC-TTRU-01-0980 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0980 gb|AAO16385.1| NADH dehydrogenase subunit 6 [Caenorhabditis elega...ns] gb|AAO16388.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16391.1| NADH dehydrogenase subunit 6 [Caenor...habditis elegans] gb|AAO16394.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AA...O16400.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16406.1| NA...DH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16409.1| NADH dehydrogenase subunit 6 [Caenorhabdi

  3. NCBI nr-aa BLAST: CBRC-TTRU-01-0496 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0496 gb|AAO16385.1| NADH dehydrogenase subunit 6 [Caenorhabditis elega...ns] gb|AAO16388.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16391.1| NADH dehydrogenase subunit 6 [Caenor...habditis elegans] gb|AAO16394.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AA...O16400.1| NADH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16406.1| NA...DH dehydrogenase subunit 6 [Caenorhabditis elegans] gb|AAO16409.1| NADH dehydrogenase subunit 6 [Caenorhabdi

  4. [Lacrimators as acceptors for NADH].

    Science.gov (United States)

    Wallenfels, K; Ertel, W; Höckendorf, A; Rieser, J; Uberschär, K H

    1975-10-01

    Lachrymators of varied structure are reduced either by hydrogen addition or halogen substitution using NADH model compounds as donors. Similar compounds without lachrymatory activity were reduced either very slowly or not at all. CS (o-Chlorobenzalmalonitril) is reduced by NADH, the reaction being catalyzed by an enzyme present in erythrocytes. Thus the lachrymatory action follows a general scheme for the activity of sensory transduction. This scheme consists of a reception in the nerve cell membrane and a fast or simultaneous chemical transformation in an enzymic reaction.

  5. NCBI nr-aa BLAST: CBRC-TTRU-01-0424 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0424 gb|ACD91639.1| NADH dehydrogenase subunit 1 [Taenia polyacantha] ...gb|ACD91640.1| NADH dehydrogenase subunit 1 [Taenia polyacantha] gb|ACD91641.1| NADH dehydrogenase subunit 1 [Taenia... polyacantha] gb|ACD91642.1| NADH dehydrogenase subunit 1 [Taenia polyacantha] gb|ACD91644.1| NADH dehydrogenase subunit 1 [Taenia polyacantha] ACD91639.1 0.037 28% ...

  6. Structural Insights into l-Tryptophan Dehydrogenase from a Photoautotrophic Cyanobacterium, Nostoc punctiforme.

    Science.gov (United States)

    Wakamatsu, Taisuke; Sakuraba, Haruhiko; Kitamura, Megumi; Hakumai, Yuichi; Fukui, Kenji; Ohnishi, Kouhei; Ashiuchi, Makoto; Ohshima, Toshihisa

    2017-01-15

    l-Tryptophan dehydrogenase from Nostoc punctiforme NIES-2108 (NpTrpDH), despite exhibiting high amino acid sequence identity (>30%)/homology (>50%) with NAD(P)(+)-dependent l-Glu/l-Leu/l-Phe/l-Val dehydrogenases, exclusively catalyzes reversible oxidative deamination of l-Trp to 3-indolepyruvate in the presence of NAD(+) Here, we determined the crystal structure of the apo form of NpTrpDH. The structure of the NpTrpDH monomer, which exhibited high similarity to that of l-Glu/l-Leu/l-Phe dehydrogenases, consisted of a substrate-binding domain (domain I, residues 3 to 133 and 328 to 343) and an NAD(+)/NADH-binding domain (domain II, residues 142 to 327) separated by a deep cleft. The apo-NpTrpDH existed in an open conformation, where domains I and II were apart from each other. The subunits dimerized themselves mainly through interactions between amino acid residues around the β-1 strand of each subunit, as was observed in the case of l-Phe dehydrogenase. The binding site for the substrate l-Trp was predicted by a molecular docking simulation and validated by site-directed mutagenesis. Several hydrophobic residues, which were located in the active site of NpTrpDH and possibly interacted with the side chain of the substrate l-Trp, were arranged similarly to that found in l-Leu/l-Phe dehydrogenases but fairly different from that of an l-Glu dehydrogenase. Our crystal structure revealed that Met-40, Ala-69, Ile-74, Ile-110, Leu-288, Ile-289, and Tyr-292 formed a hydrophobic cluster around the active site. The results of the site-directed mutagenesis experiments suggested that the hydrophobic cluster plays critical roles in protein folding, l-Trp recognition, and catalysis. Our results provide critical information for further characterization and engineering of this enzyme.

  7. Characterization of malate dehydrogenase from the hyperthermophilic archaeon Pyrobaculum islandicum.

    Science.gov (United States)

    Yennaco, Lynda J; Hu, Yajing; Holden, James F

    2007-09-01

    Native and recombinant malate dehydrogenase (MDH) was characterized from the hyperthermophilic, facultatively autotrophic archaeon Pyrobaculum islandicum. The enzyme is a homotetramer with a subunit mass of 33 kDa. The activity kinetics of the native and recombinant proteins are the same. The apparent K ( m ) values of the recombinant protein for oxaloacetate (OAA) and NADH (at 80 degrees C and pH 8.0) were 15 and 86 microM, respectively, with specific activity as high as 470 U mg(-1). Activity decreased more than 90% when NADPH was used. The catalytic efficiency of OAA reduction by P. islandicum MDH using NADH was significantly higher than that reported for any other archaeal MDH. Unlike other archaeal MDHs, specific activity of the P. islandicum MDH back-reaction also decreased more than 90% when malate and NAD(+) were used as substrates and was not detected with NADP(+). A phylogenetic tree of 31 archaeal MDHs shows that they fall into 5 distinct groups separated largely along taxonomic lines suggesting minimal lateral mdh transfer between Archaea.

  8. Structure of D-lactate dehydrogenase from Aquifex aeolicus complexed with NAD(+) and lactic acid (or pyruvate).

    Science.gov (United States)

    Antonyuk, Svetlana V; Strange, Richard W; Ellis, Mark J; Bessho, Yoshitaka; Kuramitsu, Seiki; Inoue, Yumiko; Yokoyama, Shigeyuki; Hasnain, S Samar

    2009-12-01

    The crystal structure of D-lactate dehydrogenase from Aquifex aeolicus (aq_727) was determined to 2.12 A resolution in space group P2(1)2(1)2(1), with unit-cell parameters a = 90.94, b = 94.43, c = 188.85 A. The structure was solved by molecular replacement using the coenzyme-binding domain of Lactobacillus helveticus D-lactate dehydrogenase and contained two homodimers in the asymmetric unit. Each subunit of the homodimer was found to be in a ;closed' conformation with the NADH cofactor bound to the coenzyme-binding domain and with a lactate (or pyruvate) molecule bound at the interdomain active-site cleft.

  9. The Na+-Translocating NADH:Quinone Oxidoreductase Enhances Oxidative Stress in the Cytoplasm of Vibrio cholerae.

    Science.gov (United States)

    Muras, Valentin; Dogaru-Kinn, Paul; Minato, Yusuke; Häse, Claudia C; Steuber, Julia

    2016-09-01

    We searched for a source of reactive oxygen species (ROS) in the cytoplasm of the human pathogen Vibrio cholerae and addressed the mechanism of ROS formation using the dye 2',7'-dichlorofluorescein diacetate (DCFH-DA) in respiring cells. By comparing V. cholerae strains with or without active Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR), this respiratory sodium ion redox pump was identified as a producer of ROS in vivo The amount of cytoplasmic ROS detected in V. cholerae cells producing variants of Na(+)-NQR correlated well with rates of superoxide formation by the corresponding membrane fractions. Membranes from wild-type V. cholerae showed increased superoxide production activity (9.8 ± 0.6 μmol superoxide min(-1) mg(-1) membrane protein) compared to membranes from the mutant lacking Na(+)-NQR (0.18 ± 0.01 μmol min(-1) mg(-1)). Overexpression of plasmid-encoded Na(+)-NQR in the nqr deletion strain resulted in a drastic increase in the formation of superoxide (42.6 ± 2.8 μmol min(-1) mg(-1)). By analyzing a variant of Na(+)-NQR devoid of quinone reduction activity, we identified the reduced flavin adenine dinucleotide (FAD) cofactor of cytoplasmic NqrF subunit as the site for intracellular superoxide formation in V. cholerae The impact of superoxide formation by the Na(+)-NQR on the virulence of V. cholerae is discussed. In several studies, it was demonstrated that the Na(+)-NQR in V. cholerae affects virulence in a yet unknown manner. We identified the reduced FAD cofactor in the NADH-oxidizing NqrF subunit of the Na(+)-NQR as the site of superoxide formation in the cytoplasm of V. cholerae Our study provides the framework to understand how reactive oxygen species formed during respiration could participate in the regulated expression of virulence factors during the transition from aerobic to microaerophilic (intestinal) habitats. This hypothesis may turn out to be right for many other pathogens which, like V. cholerae, depend on the Na

  10. A new dawn for plant mitochondrial NAD(P)H dehydrogenases

    DEFF Research Database (Denmark)

    Møller, I.M.

    2002-01-01

    The expression of complex I and two homologues of bacterial and yeast NADH dehydrogenases, NDA and NDB, have been studied in potato leaf mitochondria. The mRNA level of NDA is completely light dependent and shows a diurnal rhythm with a sharp maximum just after dawn. NDA protein quantity and inte...... and internal rotenone-insensitive NADH dehydrogenase activity are also light dependent. These findings suggest that NDA has a role in photorespiration and might be identical to the previously unidentified internal rotenone-insensitive NADH dehydrogenase....

  11. EST Table: FS860858 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18206.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb...|ADE18518.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18687.1| NADH dehydrogenase subunit 6 [Bombyx mandarin

  12. Identification of proteins capable of metal reduction from the proteome of the Gram-positive bacterium Desulfotomaculum reducens MI-1 using an NADH-based activity assay

    Energy Technology Data Exchange (ETDEWEB)

    Otwell, Annie E.; Sherwood, Roberts; Zhang, Sheng; Nelson, Ornella D.; Li, Zhi; Lin, Hening; Callister, Stephen J.; Richardson, Ruth E.

    2015-01-01

    Metal reduction capability has been found in numerous species of environmentally abundant Gram-positive bacteria. However, understanding of microbial metal reduction is based almost solely on studies of Gram-negative organisms. In this study, we focus on Desulfotomaculum reducens MI-1, a Gram-positive metal reducer whose genome lacks genes with similarity to any characterized metal reductase. D. reducens has been shown to reduce not only Fe(III), but also the environmentally important contaminants U(VI) and Cr(VI). By extracting, separating, and analyzing the functional proteome of D. reducens, using a ferrozine-based assay in order to screen for chelated Fe(III)-NTA reduction with NADH as electron donor, we have identified proteins not previously characterized as iron reductases. Their function was confirmed by heterologous expression in E. coli. These are the protein NADH:flavin oxidoreductase (Dred_2421) and a protein complex composed of oxidoreductase FAD/NAD(P)-binding subunit (Dred_1685) and dihydroorotate dehydrogenase 1B (Dred_1686). Dred_2421 was identified in the soluble proteome and is predicted to be a cytoplasmic protein. Dred_1685 and Dred_1686 were identified in both the soluble as well as the insoluble (presumably membrane) protein fraction, suggesting a type of membrane-association, although PSORTb predicts both proteins are cytoplasmic. Furthermore, we show that these proteins have the capability to reduce soluble Cr(VI) and U(VI) with NADH as electron donor. This study is the first functional proteomic analysis of D. reducens, and one of the first analyses of metal and radionuclide reduction in an environmentally relevant Gram-positive bacterium.

  13. Dicty_cDB: Contig-U13940-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available enase subunit F (... 40 7.9 2 ( AY331944 ) Psammisia ulbrichiana NADH dehydrogenase subunit ... 40 7.9 2 ( AY331942 ) Psammisia ecuad...orensis NADH dehydrogenase subunit... 40 7.9 2 ( AC10829

  14. Characterization of mutations in the iron-sulphur subunit of succinate dehydrogenase correlating with Boscalid resistance in Alternaria alternata from California pistachio.

    Science.gov (United States)

    Avenot, H F; Sellam, A; Karaoglanidis, G; Michailides, T J

    2008-06-01

    Thirty-eight isolates of Alternaria alternata from pistachio orchards with a history of Pristine (pyraclostrobin + boscalid) applications and displaying high levels of resistance to boscalid fungicide (mean EC(50) values >500 microg/ml) were identified following mycelial growth tests. A cross-resistance study revealed that the same isolates were also resistant to carboxin, a known inhibitor of succinate dehydrogenase (Sdh). To determine the genetic basis of boscalid resistance in A. alternata the entire iron sulphur gene (AaSdhB) was isolated from a fungicide-sensitive isolate. The deduced amino-acid sequence showed high similarity with iron sulphur proteins (Ip) from other organisms. Comparison of AaSdhB full sequences from sensitive and resistant isolates revealed that a highly conserved histidine residue (codon CAC in sensitive isolates) was converted to either tyrosine (codon TAC, type I mutants) or arginine (codon CGC, type II mutants) at position 277. In other fungal species this residue is involved in carboxamide resistance. In this study, 10 and 5 mutants were of type I and type II respectively, while 23 other resistant isolates (type III mutants) had no mutation in the histidine codon. The point mutation detected in type I mutants was used to design a pair of allele-specific polymerase chain reaction (PCR) primers to facilitate rapid detection. A PCR-restriction fragment length polymorphism (RFLP) assay in which amplified gene fragments were digested with AciI was successfully employed for the diagnosis of type II mutants. The relevance of these modifications in A. alternata AaSdhB sequence in conferring boscalid resistance is discussed.

  15. NCBI nr-aa BLAST: CBRC-ACAR-01-1147 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-1147 gb|ABW06635.1| NADH dehydrogenase subunit 2 [Anolis carolinensis]... gb|ABW06637.1| NADH dehydrogenase subunit 2 [Anolis carolinensis] gb|ABW06641.1| NADH dehydrogenase subunit 2 [Anolis caroline...nsis] gb|ABW06642.1| NADH dehydrogenase subunit 2 [Anolis carolinensis] gb|ABW06644.1| NA...DH dehydrogenase subunit 2 [Anolis carolinensis] gb|ABW06646.1| NADH dehydrogenase subunit 2 [Anolis carolinensis] ABW06635.1 0.66 25% ...

  16. Gene structure and mutations of glutaryl-coenzyme A dehydrogenase: Impaired association of enzyme subunits that is due to an A421V substitution causes glutaric acidemia type I in the Amish

    Energy Technology Data Exchange (ETDEWEB)

    Biery, B.J.; Stein, D.E.; Goodman, S.I. [Univ. of Colorado School of Medicine, Denver, CO (United States)] [and others

    1996-11-01

    The structure of the human glutaryl coenzyme A dehydrogenase (GCD) gene was determined to contain 11 exons and to span {approximately}7 kb. Fibroblast DNA from 64 unrelated glutaric academia type I (GA1) patients was screened for mutations by PCR amplification and analysis of SSCP. Fragments with altered electrophoretic mobility were subcloned and sequenced to detect mutations that caused GA1. This report describes the structure of the GCD gene, as well as point mutations and polymorphisms found in 7 of its 11 exons. Several mutations were found in more than one patient, but no one prevalent mutation was detected in the general population. As expected from pedigree analysis, a single mutant allele causes GA1 in the Old Order Amish of Lancaster County, Pennsylvania. Several mutations have been expressed in Escherichia coli, and all produce diminished enzyme activity. Reduced activity in GCD encoded by the A421V mutation in the Amish may be due to impaired association of enzyme subunits. 13 refs., 5 figs., 3 tabs.

  17. Evaluation of functioning of mitochondrial electron transport chain with NADH and FAD autofluorescence

    Directory of Open Access Journals (Sweden)

    H. V. Danylovych

    2016-02-01

    Full Text Available We prove the feasibility of evaluation of mitochondrial electron transport chain function in isolated mitochondria of smooth muscle cells of rats from uterus using fluorescence of NADH and FAD coenzymes. We found the inversely directed changes in FAD and NADH fluorescence intensity under normal functioning of mitochondrial electron transport chain. The targeted effect of inhibitors of complex I, III and IV changed fluorescence of adenine nucleotides. Rotenone (5 μM induced rapid increase in NADH fluorescence due to inhibition of complex I, without changing in dynamics of FAD fluorescence increase. Antimycin A, a complex III inhibitor, in concentration of 1 μg/ml caused sharp increase in NADH fluorescence and moderate increase in FAD fluorescence in comparison to control. NaN3 (5 mM, a complex IV inhibitor, and CCCP (10 μM, a protonophore, caused decrease in NADH and FAD fluorescence. Moreover, all the inhibitors caused mitochondria swelling. NO donors, e.g. 0.1 mM sodium nitroprusside and sodium nitrite similarly to the effects of sodium azide. Energy-dependent Ca2+ accumulation in mitochondrial matrix (in presence of oxidation substrates and Mg-ATP2- complex is associated with pronounced drop in NADH and FAD fluorescence followed by increased fluorescence of adenine nucleotides, which may be primarily due to Ca2+-dependent activation of dehydrogenases of citric acid cycle. Therefore, the fluorescent signal of FAD and NADH indicates changes in oxidation state of these nucleotides in isolated mitochondria, which may be used to assay the potential of effectors of electron transport chain.

  18. Fluorescence Lifetime Imaging of Free and Protein-Bound NADH

    Science.gov (United States)

    Lakowicz, Joseph R.; Szmacinski, Henryk; Nowaczyk, Kazimierz; Johnson, Michael L.

    1992-02-01

    We introduce a methodology, fluorescence lifetime imaging (FLIM), in which the contrast depends on the fluorescence lifetime at each point in a two-dimensional image and not on the local concentration and/or intensity of the fluorophore. We used FLIM to create lifetime images of NADH when free in solution and when bound to malate dehydrogenase. This represents a challenging case for lifetime imaging because the NADH decay times are just 0.4 and 1.0 ns in the free and bound states, respectively. In the present apparatus, lifetime images are created from a series of phase-sensitive images obtained with a gain-modulated image intensifier and recorded with a charge-coupled device (CCD) camera. The intensifier gain is modulated at the light-modulation frequency or a harmonic thereof. A series of stationary phase-sensitive images, each obtained with various phase shifts of the gain-modulation signal, is used to determine the phase angle or modulation of the emission at each pixel, which is in essence the lifetime image. We also describe an imaging procedure that allows specific decay times to be suppressed, allowing in this case suppression of the emission from either free or bound NADH. Since the fluorescence lifetimes of probes are known to be sensitive to numerous chemical and physical factors such as pH, oxygen, temperature, cations, polarity, and binding to macromolecules, this method allows imaging of the chemical or property of interest in macroscopic and microscopic samples. The concept of FLIM appears to have numerous potential applications in the biosciences.

  19. Characterization and expression of NAD(H)-dependent glutamate dehydrogenase genes in Arabidopsis.

    Science.gov (United States)

    Turano, F J; Thakkar, S S; Fang, T; Weisemann, J M

    1997-04-01

    Two distinct cDNA clones encoding NAD(H)-dependent glutamate dehydrogenase (NAD[H]-GDH) in Arabidopsis thaliana were identified and sequenced. The genes corresponding to these cDNA clones were designated GDH1 and GDH2. Analysis of the deduced amino acid sequences suggest that both gene products contain putative mitochondrial transit polypeptides and NAD(H)- and alpha-ketoglutarate-binding domains. Subcellular fractionation confirmed the mitochondrial location of the NAD(H)-GDH isoenzymes. In addition, a putative EF-hand loop, shown to be associated with Ca2+ binding, was identified in the GDH2 gene product but not in the GDH1 gene product. GDH1 encodes a 43.0-kD polypeptide, designated alpha, and GDH2 encodes a 42.5-kD polypeptide, designated beta. The two subunits combine in different ratios to form seven NAD(H)-GDH isoenzymes. The slowest-migrating isoenzyme in a native gel, GDH1, is a homohexamer composed of alpha subunits, and the fastest-migrating isoenzyme, GDH7, is a homohexamer composed of beta subunits. GDH isoenzymes 2 through 6 are heterohexamers composed of different ratios of alpha and beta subunits. NAD(H)-GDH isoenzyme patterns varied among different plant organs and in leaves of plants irrigated with different nitrogen sources or subjected to darkness for 4 d. Conversely, there were little or no measurable changes in isoenzyme patterns in roots of plants treated with different nitrogen sources. In most instances, changes in isoenzyme patterns were correlated with relative differences in the level of alpha and beta subunits. Likewise, the relative difference in the level of alpha or beta subunits was correlated with changes in the level of GDH1 or GDH2 transcript detected in each sample, suggesting that NAD(H)-GDH activity is controlled at least in part at the transcriptional level.

  20. Role of positron emission tomography and bone scintigraphy in the evaluation of bone involvement in metastatic pheochromocytoma and paraganglioma: specific implications for succinate dehydrogenase enzyme subunit B gene mutations.

    Science.gov (United States)

    Zelinka, Tomás; Timmers, Henri J L M; Kozupa, Anna; Chen, Clara C; Carrasquillo, Jorge A; Reynolds, James C; Ling, Alexander; Eisenhofer, Graeme; Lazúrová, Ivica; Adams, Karen T; Whatley, Millie A; Widimsky, Jirí; Pacak, Karel

    2008-03-01

    We performed a retrospective analysis of 71 subjects with metastatic pheochromocytoma and paraganglioma (30 subjects with mutation of succinate dehydrogenase enzyme subunit B (SDHB) gene and 41 subjects without SDHB mutation). Sixty-nine percent presented with bone metastases (SDHB +/-: 77% vs 63%), 39% with liver metastases (SDHB +/-: 27% vs 47%), and 32% with lung metastases (SDHB +/-: 37% vs 29%). The most common sites of bone involvement were thoracic spine (80%; SDHB+/-: 83% vs 77%), lumbar spine (78%; SDHB +/-: 78% vs 75%), and pelvic and sacral bones (78%; SDHB +/-: 91% vs 65%, P=0.04). Subjects with SDHB mutation also showed significantly higher involvement of long bones (SDHB +/-: 78% vs 30%, P=0.007) than those without the mutation. The best overall sensitivity in detecting bone metastases demonstrated positron emission tomography (PET) with 6-[(18)F]-fluorodopamine ([(18)F]-FDA; 90%), followed by bone scintigraphy (82%), computed tomography or magnetic resonance imaging (CT/MRI; 78%), 2-[(18)F]-fluoro-2-deoxy-d-glucose ([(18)F]-FDG) PET (76%), and scintigraphy with [(123/131)I]-metaiodobenzylguanidine (71%). In subjects with SDHB mutation, imaging modalities with best sensitivities for detecting bone metastases were CT/MRI (96%), bone scintigraphy (95%), and [(18)F]-FDG PET (92%). In subjects without SDHB mutations, the modality with the best sensitivity for bone metastases was [(18)F]-FDA PET (100%). In conclusion, bone scintigraphy should be used in the staging of patients with malignant pheochromocytoma and paraganglioma, particularly in patients with SDHB mutations. As for PET imaging, [(18)F]-FDG PET is highly recommended in SDHB mutation patients, whereas [(18)F]-FDA PET is recommended in patients without the mutation.

  1. Purification and Properties of Malate Dehydrogenase from the Extreme Thermophile Bacillus Caldolyticus

    Science.gov (United States)

    Kristjansson, Hordur; Ponnamperuma, Cyril

    1980-06-01

    The enzyme malate dehydrogenase (EC 1.1.1.37) from an extreme thermophileB. Caldolyticus was purified to about 91% homogeneity. The molar mass of the enzyme was determined as 73 000 daltons and it is composed of two subunits, each with a molar mass of 37 000. Initial velocity studies with oxaloacetic acid and NADH as substrates at pH 8.1, over a range of temperatures, indicate that the enzyme operates via a sequential type mechanism. Van't Hoff plots of the kinetic parameters displayed sharp changes in slope at characteristic temperatures, whereas the Arrhenius plot exhibited no such breaks over the temperature interval investigated. The enzyme was found to be stable at 41°C and lower temperatures. At 51°C and 59°C an almost immediate 20% reduction in activity was obtained, but no further inactivation occurred during the 60 min of incubation. At 59°C the enzyme lost 50% of its initial activity in about 38 s. High concentration of NADH was observed to greatly stabilize the enzyme at that temperature. It is suggested that the slope changes in the Van't Hoff plots and the stability profies at 51°C and 59°C are representative of a temperature induced conformational change in the enzyme.

  2. Inactivation of Lactate Dehydrogenase from Pig Heart by o-Phthalaldehyde

    Institute of Scientific and Technical Information of China (English)

    郑延斌; 王政; 陈宝玉; 王希成

    2003-01-01

    Treatment of lactate dehydrogenase (LDH) with o-phthalaldehyde resulted in a time-dependent loss of enzyme activity.The inactivation followed pseudo first-order kinetics over a wide range of the inhibitor.The second-order rate constant for the inactivation of LDH was estimated to be 1.52 (mol/L)-1·s-1.The modified enzyme showed a characteristic fluorescence emission spectrum with a maximum at 405 nm upon excitation at 337 nm, consistent with the formation of isoindole derivatives by the cross-linking of proximal cysteine and lysine residues.The loss of enzyme activity was concomitant with the increase in absorbance at 337 nm.Stoichiometric study of the reaction showed that complete loss of activity was accompanied by formation of approximately four moles of isoindole derivatives per mole of LDH subunits.One of the substrates, NADH, partially prevented the enzyme from reacting with o-phthalaldehyde, whereas the other substrate, pyruvate, did not provide any protection.Protection experiments suggest that one of the cysteine-lysine pairs modified by o-phthalaldehyde is near the NADH binding site of LDH.

  3. Characterisation of the two malate dehydrogenases from Phytomonas sp. Purification of the glycosomal isoenzyme.

    Science.gov (United States)

    Uttaro, A D; Opperdoes, F R

    1997-10-01

    Two NAD(H)-dependent malate dehydrogenase (MDH) isoenzymes were detected in Phytomonas isolated from the lactiferous tubes of Euphorbia characias. The total specific activity in crude extracts using oxaloacetate as substrate was 3.3 U mg-1 of protein. The two isoenzymes had isoelectric points of 6.0 and 7.2, respectively. The acidic isoform represented 80% of the total activity in the cell and was present in the glycosome. It was purified to homogeneity by a method involving hydrophobic interaction chromatography on Phenyl-Sepharose followed by ionic exchange on CM-Sepharose and affinity chromatography on Blue-Sepharose. The purified glycosomal MDH is a homodimeric protein with a subunit molecular mass of 37 kDa and it has a low substrate specificity, since it was able to reduce both aromatic and aliphatic alpha-ketoacids as substrate including oxaloacetate, phenyl pyruvate, alpha-keto iso-caproate and pyruvate. The apparent K(m)s for oxaloacetate and NADH were 166 and 270 microM, respectively and for L-malate and NAD+, 3000 and 246 microM, respectively. The basic isoform was present in the mitochondrion. It has a high substrate specificity and an apparent K(m) of 132 and 63 microM for oxaloacetate and NADH, respectively, and of 450 and 91 microM, respectively, with L-malate and NAD+.

  4. A fiber-optic sorbitol biosensor based on NADH fluorescence detection toward rapid diagnosis of diabetic complications.

    Science.gov (United States)

    Gessei, Tomoko; Arakawa, Takahiro; Kudo, Hiroyuki; Mitsubayashi, Kohji

    2015-09-21

    Accumulation of sorbitol in the tissue is known to cause microvascular diabetic complications. In this paper, a fiber-optic biosensor for sorbitol which is used as a biomarker of diabetic complications was developed and tested. The biosensor used a sorbitol dehydrogenase from microorganisms of the genus Flavimonas with high substrate specificity and detected the fluorescence of reduced nicotinamide adenine dinucleotide (NADH) by the enzymatic reaction. An ultraviolet light emitting diode (UV-LED) was used as the excitation light source of NADH. The fluorescence of NADH was detected using a spectrometer or a photomultiplier tube (PMT). The UV-LED and the photodetector were coupled using a Y-shaped optical fiber. In the experiment, an optical fiber probe with a sorbitol dehydrogenase immobilized membrane was placed in a cuvette filled with a phosphate buffer containing the oxidized form of nicotinamide adenine dinucleotide (NAD(+)). The changes in NADH fluorescence intensity were measured after adding a standard sorbitol solution. According to the experimental assessment, the calibration range of the sorbitol biosensor systems using a spectrometer and a PMT was 5.0-1000 μmol L(-1) and 1.0-1000 μmol L(-1), respectively. The sorbitol biosensor system using the sorbitol dehydrogenase from microorganisms of the genus Flavimonas has high selectivity and sensitivity compared with that from sheep liver. The sorbitol biosensor allows for point-of-care testing applications or daily health care tests for diabetes patients.

  5. NCBI nr-aa BLAST: CBRC-TTRU-01-0424 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0424 gb|ABD49710.1| NADH dehydrogenase subunit I [Taenia polyacantha] ...gb|ACD91632.1| NADH dehydrogenase subunit 1 [Taenia polyacantha] gb|ACD91633.1| NADH dehydrogenase subunit 1 [Taenia... polyacantha] gb|ACD91634.1| NADH dehydrogenase subunit 1 [Taenia polyacantha] gb|ACD91635.1| NADH d...ehydrogenase subunit 1 [Taenia polyacantha] gb|ACD91636.1| NADH dehydrogenase subunit 1 [Taenia... polyacantha] gb|ACD91637.1| NADH dehydrogenase subunit 1 [Taenia polyacantha] gb|ACD91638.1| NADH dehydrogenase subunit 1 [Taenia polyacantha] ABD49710.1 0.008 28% ...

  6. NCBI nr-aa BLAST: CBRC-DNOV-01-0655 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0655 gb|AAB41179.1| NADH dehydrogenase subunit 2 [Apis mellifera] gb|A...AB41181.1| NADH dehydrogenase subunit 2 [Apis mellifera] gb|AAB41183.1| NADH dehydrogenase subunit 2 [Apis mellife...ra] gb|AAB41186.1| NADH dehydrogenase subunit 2 [Apis mellifera] AAB41179.1 0.039 28% ...

  7. NCBI nr-aa BLAST: CBRC-RNOR-03-0485 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-03-0485 gb|AAY43481.1| NADH dehydrogenase subunit 2 [Milvus migrans affin...is] gb|AAY43482.1| NADH dehydrogenase subunit 2 [Milvus migrans affinis] gb|AAY43483.1| NADH dehydrogenase subunit 2 [Mil...vus migrans affinis] gb|AAY43484.1| NADH dehydrogenase subunit 2 [Milvus migrans affinis] AAY43481.1 0.32 25% ...

  8. White shrimp Litopenaeus vannamei recombinant lactate dehydrogenase: Biochemical and kinetic characterization.

    Science.gov (United States)

    Fregoso-Peñuñuri, Ambar A; Valenzuela-Soto, Elisa M; Figueroa-Soto, Ciria G; Peregrino-Uriarte, Alma B; Ochoa-Valdez, Manuel; Leyva-Carrillo, Lilia; Yepiz-Plascencia, Gloria

    2017-09-01

    Shrimp lactate dehydrogenase (LDH) is induced in response to environmental hypoxia. Two protein subunits deduced from different transcripts of the LDH gene from the shrimp Litopenaeus vannamei (LDHvan-1 and LDHvan-2) were identified. These subunits are expressed by alternative splicing. Since both subunits are expressed in most tissues, the purification of the enzyme from the shrimp will likely produce hetero LDH containing both subunits. Therefore, the aim of this study was to overexpress, purify and characterize only one subunit as a recombinant protein, the LDHvan-2. For this, the cDNA from muscle was cloned and overexpressed in E. coli as a fusion protein containing an intein and a chitin binding protein domain (CBD). The recombinant protein was purified by chitin affinity chromatography column that retained the CBD and released solely the full and active LDH. The active protein appears to be a tetramer with molecular mass of approximately 140 kDa and can use pyruvate or lactate as substrates, but has higher specific activity with pyruvate. The enzyme is stable between pH 7.0 to 8.5, and between 20 and 50 °C with an optimal temperature of 50 °C. Two pKa of 9.3 and 6.6, and activation energy of 44.8 kJ/mol°K were found. The kinetic constants Km for NADH was 23.4 ± 1.8 μM, and for pyruvate was 203 ± 25 μM, while Vmax was 7.45 μmol/min/mg protein. The shrimp LDH that is mainly expressed in shrimp muscle preferentially converts pyruvate to lactate and is an important enzyme for the response to hypoxia. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Application of NAD-dependent polyol dehydrogenases for enzymatic mannitol/sorbitol production with coenzyme regeneration.

    Science.gov (United States)

    Parmentier, S; Arnaut, F; Soetaert, W; Vandamme, E J

    2003-01-01

    D-Mannitol and D-sorbitol were produced enzymatically from D-fructose using NAD-dependent polyol dehydrogenases. For the production of D-mannitol the Leuconostoc mesenteroides mannitol dehydrogenase could be used. Gluconobacter oxydans cell extract contained however both mannitol and sorbitol dehydrogenase. When this cell extract was used, the reduction of D-fructose resulted in a mixture of D-sorbitol and D-mannitol. To determine the optimal bioconversion conditions the polyol dehydrogenases were characterized towards pH- and temperature-optimum and -stability. As a compromise between enzyme activity and stability, the bioconversion reactions were performed at pH 6.5 and 25 degrees C. Since the polyol dehydrogenases are NADH-dependent, an efficient coenzyme regeneration was needed. Regeneration of NADH was accomplished by formate dehydrogenase-mediated oxidation of formate into CO2.

  10. Free [NADH]/[NAD(+)] regulates sirtuin expression.

    Science.gov (United States)

    Gambini, Juan; Gomez-Cabrera, Mari Carmen; Borras, Consuelo; Valles, Soraya L; Lopez-Grueso, Raul; Martinez-Bello, Vladimir E; Herranz, Daniel; Pallardo, Federico V; Tresguerres, Jesus A F; Serrano, Manuel; Viña, Jose

    2011-08-01

    Sirtuins are deacetylases involved in metabolic regulation and longevity. Our aim was to test the hypothesis that they are subjected to redox regulation by the [NADH]/[NAD(+)] ratio. We used NIH3T3 fibroblasts in culture, Drosophila fed with or without ethanol and exercising rats. In all three models an increase in [NADH]/[NAD(+)] came up with an increased expression of sirtuin mRNA and protein. PGC-1α (a substrate of sirtuins) protein level was significantly increased in fibroblasts incubated with lactate and pyruvate but this effect was lost in fibroblasts obtained from sirtuin-deficient mice. We conclude that the expression of sirtuins is subject to tight redox regulation by the [NADH]/[NAD(+)] ratio, which is a major sensor for metabolite availability conserved from invertebrates to vertebrates. Copyright © 2011. Published by Elsevier Inc.

  11. NCBI nr-aa BLAST: CBRC-TTRU-01-1275 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1275 ref|YP_001315040.1| NADH dehydrogenase subunit 4 [Litopenaeus van...namei] gb|ABF58013.1| NADH dehydrogenase subunit 4 [Litopenaeus vannamei] gb|ABQ84185.1| NADH dehydrogenase subunit 4 [Litopenaeus vannamei] YP_001315040.1 0.015 29% ...

  12. NCBI nr-aa BLAST: CBRC-TTRU-01-0711 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0711 ref|YP_001382117.1| NADH dehydrogenase subunit 6 [Fenneropenaeus ...chinensis] gb|ABF83979.1| NADH dehydrogenase subunit 6 [Fenneropenaeus chinensis] gb|ABG65675.1| NADH dehydrogenase subunit 6 [Fenneropenaeus chinensis] YP_001382117.1 0.006 31% ...

  13. NCBI nr-aa BLAST: CBRC-FCAT-01-0220 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-FCAT-01-0220 ref|YP_514772.1| NADH dehydrogenase subunit 6 [Watasenia scintill...ans] dbj|BAD52103.1| NADH dehydrogenase subunit 6 [Watasenia scintillans] dbj|BAE80020.1| NADH dehydrogenase subunit 6 [Watasenia scintillans] YP_514772.1 0.39 27% ...

  14. Gene : CBRC-TTRU-01-1020 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available ksi] ref|YP_271924.1| NADH dehydrogenase subunit 5 [Montastraea annularis] dbj|BAE16177.1| NADH dehydrogenas...e subunit 5 [Montastraea annularis] dbj|BAE16190.1| NADH dehydrogenase subunit 5 [Montastraea annular

  15. NCBI nr-aa BLAST: CBRC-RNOR-03-0485 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-03-0485 gb|AAY43499.1| NADH dehydrogenase subunit 2 [Milvus migrans parasit...us] gb|AAY43500.1| NADH dehydrogenase subunit 2 [Milvus migrans parasitus] gb|AAY43501.1| NADH dehydrogenase subunit 2 [Milvus migrans parasitus] AAY43499.1 0.32 25% ...

  16. NCBI nr-aa BLAST: CBRC-TTRU-01-0110 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0110 gb|ACF49099.1| NADH dehydrogenase subunit 1 [Medousogyliauchen pa...nope] gb|ACF49100.1| NADH dehydrogenase subunit 1 [Medousogyliauchen panope] gb|ACF49101.1| NADH dehydrogenase subunit 1 [Medousogyliauchen panope] ACF49099.1 0.084 30% ...

  17. NCBI nr-aa BLAST: CBRC-PABE-13-0076 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PABE-13-0076 ref|YP_073334.1| NADH dehydrogenase subunit 1 [Aleurochiton aceri...s] gb|AAS77045.1| NADH dehydrogenase subunit 1 [Aleurochiton aceris] gb|AAS77789.1| NADH dehydrogenase subunit 1 [Aleurochiton aceris] YP_073334.1 0.006 22% ...

  18. NCBI nr-aa BLAST: CBRC-DNOV-01-0670 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0670 gb|AAC98238.1| NADH dehydrogenase subunit 4 [Haemonchus placei] g...b|AAC98239.1| NADH dehydrogenase subunit 4 [Haemonchus placei] gb|AAC98243.1| NADH dehydrogenase subunit 4 [Haemonchus placei] AAC98238.1 1.7 36% ...

  19. NCBI nr-aa BLAST: CBRC-TTRU-01-1312 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1312 gb|AAC98212.1| NADH dehydrogenase subunit 4 [Haemonchus placei] g...b|AAC98215.1| NADH dehydrogenase subunit 4 [Haemonchus placei] gb|AAC98218.1| NADH dehydrogenase subunit 4 [Haemonchus placei] AAC98212.1 0.014 31% ...

  20. NAD+-dependent glutamate dehydrogenase of the edible mushroom Agaricus bisporus: biochemical and molecular characterization.

    Science.gov (United States)

    Kersten, M A; Müller, Y; Baars, J J; Op den Camp, H J; van der Drift, C; Van Griensven, L J; Visser, J; Schaap, P J

    1999-04-01

    The NAD+-dependent glutamate dehydrogenase (NAD-GDH) of Agaricus bisporus, a key enzyme in nitrogen metabolism, was purified to homogeneity. The apparent molecular mass of the native enzyme is 474 kDa comprising four subunits of 116 kDa. The isoelectric point of the enzyme is about 7.0. Km values for ammonium, 2-oxoglutarate, NADH, glutamate and NAD+ were 6.5, 3.5, 0.06, 37.1 and 0.046 mM, respectively. The enzyme is specific for NAD(H). The gene encoding this enzyme (gdhB) was isolated from an A. bisporus H39 recombinant lambda phage library. The deduced amino acid sequence specifies a 1029-amino acid protein with a deduced molecular mass of 115,463 Da, which displays a significant degree of similarity with NAD-GDH of Saccharomyces cerevisiae and Neurospora crassa. The ORF is interrupted by fifteen introns. Northern analysis combined with enzyme activity measurements suggest that NAD-GDH from A. bisporus is regulated by the nitrogen source. NAD-GDH levels in mycelium grown on glutamate were higher than NAD-GDH levels in mycelium grown on ammonium as a nitrogen source. Combined with the kinetic parameters, these results suggest a catabolic role for NAD-GDH. However, upon addition of ammonium to the culture transcription of the gene is not repressed as strongly as that of the gene encoding NADP-GDH (gdhA). To date, tetrameric NAD-GDHs with large subunits, and their corresponding genes, have only been isolated from a few species. This enzyme represents the first NAD-GDH of basidiomycete origin to be purified and is the first such enzyme from basidiomycetes whose sequence has been determined.

  1. Purification and characterization of benzyl alcohol- and benzaldehyde- dehydrogenase from Pseudomonas putida CSV86.

    Science.gov (United States)

    Shrivastava, Rahul; Basu, Aditya; Phale, Prashant S

    2011-08-01

    Pseudomonas putida CSV86 utilizes benzyl alcohol via catechol and methylnaphthalenes through detoxification pathway via hydroxymethylnaphthalenes and naphthaldehydes. Based on metabolic studies, benzyl alcohol dehydrogenase (BADH) and benzaldehyde dehydrogenase (BZDH) were hypothesized to be involved in the detoxification pathway. BADH and BZDH were purified to apparent homogeneity and were (1) homodimers with subunit molecular mass of 38 and 57 kDa, respectively, (2) NAD(+) dependent, (3) broad substrate specific accepting mono- and di-aromatic alcohols and aldehydes but not aliphatic compounds, and (4) BADH contained iron and magnesium, while BZDH contained magnesium. BADH in the forward reaction converted alcohol to aldehyde and required NAD(+), while in the reverse reaction it reduced aldehyde to alcohol in NADH-dependent manner. BZDH showed low K (m) value for benzaldehyde as compared to BADH reverse reaction. Chemical cross-linking studies revealed that BADH and BZDH do not form multi-enzyme complex. Thus, the conversion of aromatic alcohol to acid is due to low K (m) and high catalytic efficiency of BZDH. Phylogenetic analysis revealed that BADH is a novel enzyme and diverged during the evolution to gain the ability to utilize mono- and di-aromatic compounds. The wide substrate specificity of these enzymes enables strain to detoxify methylnaphthalenes to naphthoic acids efficiently.

  2. Mutational analysis of the hyc-operon determining the relationship between hydrogenase-3 and NADH pathway in Enterobacter aerogenes.

    Science.gov (United States)

    Pi, Jian; Jawed, Muhammad; Wang, Jun; Xu, Li; Yan, Yunjun

    2016-01-01

    In this study, the hydrogenase-3 gene cluster (hycDEFGH) was isolated and identified from Enterobacter aerogenes CCTCC AB91102. All gene products were highly homologous to the reported bacterial hydrogenase-3 (Hyd-3) proteins. The genes hycE, hycF, hycG encoding the subunits of hydrogenase-3 were targeted for genetic knockout to inhibit the FHL hydrogen production pathway via the Red recombination system, generating three mutant strains AB91102-E (ΔhycE), AB91102-F (ΔhycF) and AB91102-G (ΔhycG). Deletion of the three genes affected the integrity of hydrogenase-3. The hydrogen production experiments with the mutant strains showed that no hydrogen was detected compared with the wild type (0.886 mol/mol glucose), demonstrating that knocking out any of the three genes could inhibit NADH hydrogen production pathway. Meanwhile, the metabolites of the mutant strains were significantly changed in comparison with the wild type, indicating corresponding changes in metabolic flux by mutation. Additionally, the activity of NADH-mediated hydrogenase was found to be nil in the mutant strains. The chemostat experiments showed that the NADH/NAD(+) ratio of the mutant strains increased nearly 1.4-fold compared with the wild type. The NADH-mediated hydrogenase activity and NADH/NAD(+) ratio analysis both suggested that NADH pathway required the involvement of the electron transport chain of hydrogenase-3.

  3. Impact of overexpressing NADH kinase on glucose and xylose metabolism in recombinant xylose-utilizing Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Hou, Jin; Vemuri, G. N.; Bao, X. M.;

    2009-01-01

    During growth of Saccharomyces cerevisiae on glucose, the redox cofactors NADH and NADPH are predominantly involved in catabolism and biosynthesis, respectively. A deviation from the optimal level of these cofactors often results in major changes in the substrate uptake and biomass formation....... However, the metabolism of xylose by recombinant S. cerevisiae carrying xylose reductase and xylitol dehydrogenase from the fungal pathway requires both NADH and NADPH and creates cofactor imbalance during growth on xylose. As one possible solution to overcoming this imbalance, the effect...... in the cytosol redirected carbon flow from CO2 to ethanol during aerobic growth on glucose and to ethanol and acetate during anaerobic growth on glucose. However, cytosolic NADH kinase has an opposite effect during anaerobic metabolism of xylose consumption by channeling carbon flow from ethanol to xylitol...

  4. Characterization of water-forming NADH oxidases for co-factor regeneration

    DEFF Research Database (Denmark)

    Rehn, Gustav; Pedersen, Asbjørn Toftgaard; J. Charnock, Simon

    an environmentaland economic perspective [1]. Alcohol dehydrogenases (ADH) offer one such alternative. However, the reaction requires the oxidized nicotinamide co-factor (NAD+) that must be recycled due to its high cost contribution. One regeneration method that offers certain advantages is the oxidation of NADH...... using water forming NADH oxidases (NOX-2). The implementation of the ADH/NOX system for alcohol oxidation, however, requires consideration of several different issues. Enzyme activity and stability at relevant pH and temperature conditions, but also the tolerance to the substrates and products present......Traditional chemical methods for alcohol oxidation are often associated with issues such as high consumption of expensive oxidizing agents, generation of metal waste and the use of environmentally undesirable organic solvents. Developing green, selective catalysts is therefore important from...

  5. Structural basis of cooperativity in human UDP-glucose dehydrogenase.

    Directory of Open Access Journals (Sweden)

    Venkatachalam Rajakannan

    Full Text Available BACKGROUND: UDP-glucose dehydrogenase (UGDH is the sole enzyme that catalyzes the conversion of UDP-glucose to UDP-glucuronic acid. The product is used in xenobiotic glucuronidation in hepatocytes and in the production of proteoglycans that are involved in promoting normal cellular growth and migration. Overproduction of proteoglycans has been implicated in the progression of certain epithelial cancers, while inhibition of UGDH diminished tumor angiogenesis in vivo. A better understanding of the conformational changes occurring during the UGDH reaction cycle will pave the way for inhibitor design and potential cancer therapeutics. METHODOLOGY: Previously, the substrate-bound of UGDH was determined to be a symmetrical hexamer and this regular symmetry is disrupted on binding the inhibitor, UDP-α-D-xylose. Here, we have solved an alternate crystal structure of human UGDH (hUGDH in complex with UDP-glucose at 2.8 Å resolution. Surprisingly, the quaternary structure of this substrate-bound protein complex consists of the open homohexamer that was previously observed for inhibitor-bound hUGDH, indicating that this conformation is relevant for deciphering elements of the normal reaction cycle. CONCLUSION: In all subunits of the present open structure, Thr131 has translocated into the active site occupying the volume vacated by the absent active water and partially disordered NAD+ molecule. This conformation suggests a mechanism by which the enzyme may exchange NADH for NAD+ and repolarize the catalytic water bound to Asp280 while protecting the reaction intermediates. The structure also indicates how the subunits may communicate with each other through two reaction state sensors in this highly cooperative enzyme.

  6. Origin and evolution of the sodium -pumping NADH: ubiquinone oxidoreductase.

    Science.gov (United States)

    Reyes-Prieto, Adrian; Barquera, Blanca; Juárez, Oscar

    2014-01-01

    The sodium -pumping NADH: ubiquinone oxidoreductase (Na+-NQR) is the main ion pump and the primary entry site for electrons into the respiratory chain of many different types of pathogenic bacteria. This enzymatic complex creates a transmembrane gradient of sodium that is used by the cell to sustain ionic homeostasis, nutrient transport, ATP synthesis, flagellum rotation and other essential processes. Comparative genomics data demonstrate that the nqr operon, which encodes all Na+-NQR subunits, is found in a large variety of bacterial lineages with different habitats and metabolic strategies. Here we studied the distribution, origin and evolution of this enzymatic complex. The molecular phylogenetic analyses and the organizations of the nqr operon indicate that Na+-NQR evolved within the Chlorobi/Bacteroidetes group, after the duplication and subsequent neofunctionalization of the operon that encodes the homolog RNF complex. Subsequently, the nqr operon dispersed through multiple horizontal transfer events to other bacterial lineages such as Chlamydiae, Planctomyces and α, β, γ and δ -proteobacteria. Considering the biochemical properties of the Na+-NQR complex and its physiological role in different bacteria, we propose a detailed scenario to explain the molecular mechanisms that gave rise to its novel redox- dependent sodium -pumping activity. Our model postulates that the evolution of the Na+-NQR complex involved a functional divergence from its RNF homolog, following the duplication of the rnf operon, the loss of the rnfB gene and the recruitment of the reductase subunit of an aromatic monooxygenase.

  7. Origin and evolution of the sodium -pumping NADH: ubiquinone oxidoreductase.

    Directory of Open Access Journals (Sweden)

    Adrian Reyes-Prieto

    Full Text Available The sodium -pumping NADH: ubiquinone oxidoreductase (Na+-NQR is the main ion pump and the primary entry site for electrons into the respiratory chain of many different types of pathogenic bacteria. This enzymatic complex creates a transmembrane gradient of sodium that is used by the cell to sustain ionic homeostasis, nutrient transport, ATP synthesis, flagellum rotation and other essential processes. Comparative genomics data demonstrate that the nqr operon, which encodes all Na+-NQR subunits, is found in a large variety of bacterial lineages with different habitats and metabolic strategies. Here we studied the distribution, origin and evolution of this enzymatic complex. The molecular phylogenetic analyses and the organizations of the nqr operon indicate that Na+-NQR evolved within the Chlorobi/Bacteroidetes group, after the duplication and subsequent neofunctionalization of the operon that encodes the homolog RNF complex. Subsequently, the nqr operon dispersed through multiple horizontal transfer events to other bacterial lineages such as Chlamydiae, Planctomyces and α, β, γ and δ -proteobacteria. Considering the biochemical properties of the Na+-NQR complex and its physiological role in different bacteria, we propose a detailed scenario to explain the molecular mechanisms that gave rise to its novel redox- dependent sodium -pumping activity. Our model postulates that the evolution of the Na+-NQR complex involved a functional divergence from its RNF homolog, following the duplication of the rnf operon, the loss of the rnfB gene and the recruitment of the reductase subunit of an aromatic monooxygenase.

  8. Sources and implications of NADH/NAD+ redox imbalance in diabetes and its complications

    Directory of Open Access Journals (Sweden)

    Wu J

    2016-05-01

    Full Text Available Jinzi Wu,1Zhen Jin,1Hong Zheng,1,2Liang-Jun Yan1 1Department of Pharmaceutical Sciences, UNT System College of Pharmacy, University of North Texas Health Science Center, Fort Worth, TX, USA; 2Department of Basic Theory of Traditional Chinese Medicine, College of Basic Medicine, Shandong University of Traditional Chinese Medicine, Jinan, People’s Republic of China Abstract: NAD+ is a fundamental molecule in metabolism and redox signaling. In diabetes and its complications, the balance between NADH and NAD+ can be severely perturbed. On one hand, NADH is overproduced due to influx of hyperglycemia to the glycolytic and Krebs cycle pathways and activation of the polyol pathway. On the other hand, NAD+ can be diminished or depleted by overactivation of poly ADP ribose polymerase that uses NAD+ as its substrate. Moreover, sirtuins, another class of enzymes that also use NAD+ as their substrate for catalyzing protein deacetylation reactions, can also affect cellular content of NAD+. Impairment of NAD+ regeneration enzymes such as lactate dehydrogenase in erythrocytes and complex I in mitochondria can also contribute to NADH accumulation and NAD+ deficiency. The consequence of NADH/NAD+ redox imbalance is initially reductive stress that eventually leads to oxidative stress and oxidative damage to macromolecules, including DNA, lipids, and proteins. Accordingly, redox imbalance-triggered oxidative damage has been thought to be a major factor contributing to the development of diabetes and its complications. Future studies on restoring NADH/NAD+ redox balance could provide further insights into design of novel antidiabetic strategies. Keywords: mitochondria, complex I, reactive oxygen species, polyol pathway, poly ADP ribosylation, sirtuins, oxidative stress, oxidative damage

  9. Increased availability of NADH in metabolically engineered baker's yeast improves transaminase-oxidoreductase coupled asymmetric whole-cell bioconversion

    DEFF Research Database (Denmark)

    Knudsen, Jenny Dahl; Hägglöf, Cecilia; Weber, Nora

    2016-01-01

    yeast for transamination-reduction coupled asymmetric one-pot conversion was investigated. RESULTS: A series of active whole-cell biocatalysts were constructed by over-expressing the (S)-selective ω-transaminase (VAMT) from Capsicum chinense together with the NADH-dependent (S)-selective alcohol...... dehydrogenase (SADH) originating from Rhodococcus erythropolis in strains with or without deletion of glycerol-3-phosphate dehydrogenases 1 and 2 (GPD1 and GPD2). The yeast strains were evaluated as catalysts for simultaneous: (a) kinetic resolution of the racemic mixture to (R)-1-phenylethylamine, and (b...

  10. Cold stress decreases the capacity for respiratory NADH oxidation in potato leaves

    DEFF Research Database (Denmark)

    Svensson, Å.S.; Johansson, F.I.; Møller, I.M.

    2002-01-01

    Cold stress effects on the expression of genes for respiratory chain enzymes were investigated in potato (Solarium tuberosum L., cv. Desiree) leaves. The nda1 and ndb1 genes, homologues to genes encoding the non-proton-pumping respiratory chain NADH dehydrogenases of Escherichia coli and yeast, w....... The results are discussed in relation to the recent finding that the nda1 gene expression is completely light-dependent. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved....

  11. Purification and properties of three NAD(P)+ isozymes of L-glutamate dehydrogenase of Chlamydomonas reinhardtii.

    Science.gov (United States)

    Moyano, E; Cárdenas, J; Muñoz-Blanco, J

    1992-02-13

    Three isozymes of glutamate dehydrogenase (GDH) of Chlamydomonas reinhardtii, induced under different trophic and stress conditions, have been purified about 800-1000-fold to electrophoretic homogeneity. They are hexamers of Mr 266,000-269,000 as deduced from gel filtration and sedimentation coefficient data. GDH1 consisted of six identical subunits of 44 kDa each, whereas both GDH2 and GDH3 consisted of six similar-sized monomers (4 of 44 kDa and 2 of 46 kDa). Optimum pH for the three activities with each pyridine nucleotide was identical (8.5 with NADH; 7.7 with NADPH; and 9.0 with NAD+). The isozymes exhibited similar high optimum temperature values (60-62 degrees C) and isoelectric points (7.9-8.1). Activity was enhanced in vitro by Ca2+ ions and strongly inhibited by pyridoxal 5'-phosphate, KCN, o-phenanthroline and EDTA, and to a lesser extent by pHMB and methylacetimidate. In the aminating reaction the three isozymes were inhibited in a concentration-dependent process by both NADH and NADPH, with apparent Km values for NH4+ ranging from 13-53 mM; 0.36-1.85 mM for 2-oxoglutarate and 0.07-0.78 mM for NADH and NADPH. In the deaminating reaction apparent Km values ranged from 0.64-3.52 mM for L-glutamate and 0.20-0.32 for NAD+. In addition, the three isozymes exhibited a non-hyperbolic kinetics for NAD+ with negative cooperativity (n = 0.8).

  12. Dicty_cDB: Contig-U13969-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available tia... 48 0.79 1 ( AY083455 ) Melanoplus bowditchi NADH dehydrogenase subunit I... 48 0.79 1 ( AY083411 ) Melanoplus infantil...is isolate 4 NADH dehydrogenas... 48 0.79 1 ( AY083407 ) Melanoplus infantilis isolate 3 NA...hydrogenase subunit II... 48 0.79 1 ( AF317154 ) Melanoplus infantilis NADH dehydrogenase subunit ... 48 0.7....79 1 ( AF227281 ) Melanoplus infantilis NADH dehydrogenase subunit ... 48 0.79 1 ( AC113352 ) Homo sapiens

  13. Structural characterization of tartrate dehydrogenase: a versatile enzyme catalyzing multiple reactions

    Energy Technology Data Exchange (ETDEWEB)

    Malik, Radhika; Viola, Ronald E. (Toledo)

    2010-10-28

    The first structure of an NAD-dependent tartrate dehydrogenase (TDH) has been solved to 2 {angstrom} resolution by single anomalous diffraction (SAD) phasing as a complex with the intermediate analog oxalate, Mg{sup 2+} and NADH. This TDH structure from Pseudomonas putida has a similar overall fold and domain organization to other structurally characterized members of the hydroxy-acid dehydrogenase family. However, there are considerable differences between TDH and these functionally related enzymes in the regions connecting the core secondary structure and in the relative positioning of important loops and helices. The active site in these complexes is highly ordered, allowing the identification of the substrate-binding and cofactor-binding groups and the ligands to the metal ions. Residues from the adjacent subunit are involved in both the substrate and divalent metal ion binding sites, establishing a dimer as the functional unit and providing structural support for an alternating-site reaction mechanism. The divalent metal ion plays a prominent role in substrate binding and orientation, together with several active-site arginines. Functional groups from both subunits form the cofactor-binding site and the ammonium ion aids in the orientation of the nicotinamide ring of the cofactor. A lysyl amino group (Lys192) is the base responsible for the water-mediated proton abstraction from the C2 hydroxyl group of the substrate that begins the catalytic reaction, followed by hydride transfer to NAD. A tyrosyl hydroxyl group (Tyr141) functions as a general acid to protonate the enolate intermediate. Each substrate undergoes the initial hydride transfer, but differences in substrate orientation are proposed to account for the different reactions catalyzed by TDH.

  14. The NADH oxidase-Prx system in Amphibacillus xylanus.

    Science.gov (United States)

    Niimura, Youichi

    2007-01-01

    Amphibacillus NADH oxidase belongs to a growing new family of peroxiredoxin-linked oxidoreductases including alkyl hydroperoxide reductase F (AhpF). Like AhpF it displays extremely high hydroperoxide reductase activity in the presence of a Prx, thus making up the NADH oxidase-Prx system. The NADH oxidase primarily catalyzes the reduction of oxygen by NADH to form H2O2, while the Prx immediately reduces H2O2 (or ROOH) to water (or ROH). Consequently, the NADH oxidase-Prx system catalyzes the reduction of both oxygen and hydrogen peroxide to water with NADH as the preferred electron donor. The NADH oxidase-Prx system is widely distributed in aerobically growing bacteria lacking a respiratory chain and catalase, and plays an important role not only in scavenging hydroperoxides but also in regenerating NAD in these bacteria.

  15. Cloning and sequence analysis of the gene encoding 19-kD subunit of Complex I from Dunaliella salina.

    Science.gov (United States)

    Liu, Yi; Qiao, Dai Rong; Zheng, Hong Bo; Dai, Xu Lan; Bai, Lin Han; Zeng, Jing; Cao, Yi

    2008-09-01

    NADH:ubiquinone oxidoreductase (complex I ) of the mitochondrial respiratory chain catalyzes the transfer of electrons from NADH to ubiquinone coupled to proton translocation across the membrane. The cDNA sequence of Dunaliella salina mitochondrial NADH: ubiquinone oxidoreductase 19-kD subunit contains a 682-bp ORF encoding a protein with an apparent molecular mass of 19 kD. The sequence has been submitted to the GenBank database under Accession No. EF566890 (cDNA sequences) and EF566891 (genomic sequence). The deduced amino-acid sequence is 74% identical to Chlamydomonas reinhardtii mitochondrial NADH:ubiquinone oxidoreductase 18-kD subunit. The 19-kD subunit mRNA expression was observed in oxygen deficiency, salt treatment, and rotenone treatment with lower levels. It demonstrate that the 19-kD subunit of Complex I from Dunaliella salina is regulated by these stresses.

  16. Increasing anaerobic acetate consumption and ethanol yields in Saccharomyces cerevisiae with NADPH-specific alcohol dehydrogenase.

    Science.gov (United States)

    Henningsen, Brooks M; Hon, Shuen; Covalla, Sean F; Sonu, Carolina; Argyros, D Aaron; Barrett, Trisha F; Wiswall, Erin; Froehlich, Allan C; Zelle, Rintze M

    2015-12-01

    Saccharomyces cerevisiae has recently been engineered to use acetate, a primary inhibitor in lignocellulosic hydrolysates, as a cosubstrate during anaerobic ethanolic fermentation. However, the original metabolic pathway devised to convert acetate to ethanol uses NADH-specific acetylating acetaldehyde dehydrogenase and alcohol dehydrogenase and quickly becomes constrained by limited NADH availability, even when glycerol formation is abolished. We present alcohol dehydrogenase as a novel target for anaerobic redox engineering of S. cerevisiae. Introduction of an NADPH-specific alcohol dehydrogenase (NADPH-ADH) not only reduces the NADH demand of the acetate-to-ethanol pathway but also allows the cell to effectively exchange NADPH for NADH during sugar fermentation. Unlike NADH, NADPH can be freely generated under anoxic conditions, via the oxidative pentose phosphate pathway. We show that an industrial bioethanol strain engineered with the original pathway (expressing acetylating acetaldehyde dehydrogenase from Bifidobacterium adolescentis and with deletions of glycerol-3-phosphate dehydrogenase genes GPD1 and GPD2) consumed 1.9 g liter(-1) acetate during fermentation of 114 g liter(-1) glucose. Combined with a decrease in glycerol production from 4.0 to 0.1 g liter(-1), this increased the ethanol yield by 4% over that for the wild type. We provide evidence that acetate consumption in this strain is indeed limited by NADH availability. By introducing an NADPH-ADH from Entamoeba histolytica and with overexpression of ACS2 and ZWF1, we increased acetate consumption to 5.3 g liter(-1) and raised the ethanol yield to 7% above the wild-type level. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  17. Engineering Pichia pastoris for improved NADH regeneration: A novel chassis strain for whole-cell catalysis

    Directory of Open Access Journals (Sweden)

    Martina Geier

    2015-09-01

    Full Text Available Many synthetically useful reactions are catalyzed by cofactor-dependent enzymes. As cofactors represent a major cost factor, methods for efficient cofactor regeneration are required especially for large-scale synthetic applications. In order to generate a novel and efficient host chassis for bioreductions, we engineered the methanol utilization pathway of Pichia pastoris for improved NADH regeneration. By deleting the genes coding for dihydroxyacetone synthase isoform 1 and 2 (DAS1 and DAS2, NADH regeneration via methanol oxidation (dissimilation was increased significantly. The resulting Δdas1 Δdas2 strain performed better in butanediol dehydrogenase (BDH1 based whole-cell conversions. While the BDH1 catalyzed acetoin reduction stopped after 2 h reaching ~50% substrate conversion when performed in the wild type strain, full conversion after 6 h was obtained by employing the knock-out strain. These results suggest that the P. pastoris Δdas1 Δdas2 strain is capable of supplying the actual biocatalyst with the cofactor over a longer reaction period without the over-expression of an additional cofactor regeneration system. Thus, focusing the intrinsic carbon flux of this methylotrophic yeast on methanol oxidation to CO2 represents an efficient and easy-to-use strategy for NADH-dependent whole-cell conversions. At the same time methanol serves as co-solvent, inductor for catalyst and cofactor regeneration pathway expression and source of energy.

  18. EST Table: FS737382 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  19. EST Table: FS872671 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fner ...

  20. EST Table: BY914601 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/29 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h DN237205 an-- ...

  1. EST Table: FS744614 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/07 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  2. EST Table: BY922710 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/29 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 ovS0 ...

  3. EST Table: FS737591 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  4. EST Table: FS731263 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  5. EST Table: FS791098 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 ffbm ...

  6. EST Table: FS724700 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  7. EST Table: BY931227 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/29 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 ovS0 ...

  8. EST Table: FS734614 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  9. EST Table: FS835928 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/10 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fmgV ...

  10. EST Table: FS853128 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fner ...

  11. EST Table: BB986940 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/28 n.h 10/08/27 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 MSV3 ...

  12. EST Table: FS842132 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/10 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fner ...

  13. EST Table: FS778806 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/08 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fcaL ...

  14. EST Table: FS868644 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 30 %/152 aa FBpp0100185|mt:ND6-PA 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fner ...

  15. EST Table: FS808527 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fmgV ...

  16. EST Table: FS737834 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  17. EST Table: FS877704 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 ftes ...

  18. EST Table: FS729860 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  19. EST Table: FS792253 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 ffbm ...

  20. EST Table: FS771359 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/08 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fcaL ...

  1. EST Table: BY937322 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h DN237205 prW- ...

  2. EST Table: FS735238 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  3. EST Table: BB992901 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 PSV3 ...

  4. EST Table: FS815084 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fmgV ...

  5. EST Table: FS730426 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  6. EST Table: FS816925 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fmgV ...

  7. EST Table: FS803907 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fmgV ...

  8. EST Table: BB989559 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h DN237205 PSV3 ...

  9. EST Table: FS850307 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fner ...

  10. EST Table: FS838622 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/10 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fner ...

  11. EST Table: FS734674 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  12. EST Table: FS854365 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fner ...

  13. EST Table: FS853563 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fner ...

  14. EST Table: BY931776 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/29 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 ovS0 ...

  15. EST Table: FS741005 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/07 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  16. EST Table: FS849800 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fner ...

  17. EST Table: FS740649 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/07 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  18. EST Table: FS914429 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/12 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fufe ...

  19. EST Table: FS751581 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/08 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 caL- ...

  20. EST Table: FS920744 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/13 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fufe ...

  1. EST Table: FS726233 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  2. EST Table: FS830829 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/10 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fmgV ...

  3. EST Table: FS857377 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fner ...

  4. EST Table: FS871451 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fner ...

  5. EST Table: FS740442 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/07 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  6. EST Table: FS842158 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/10 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fner ...

  7. EST Table: FS761870 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/08 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fcaL ...

  8. EST Table: FS868155 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fner ...

  9. EST Table: FS727278 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  10. EST Table: BB992059 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/28 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 MSV3 ...

  11. EST Table: FS744551 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/07 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  12. EST Table: FS734081 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  13. EST Table: FS736778 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  14. EST Table: FS727040 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  15. EST Table: FS726833 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  16. EST Table: FS827516 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/10 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fmgV ...

  17. EST Table: FS811846 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fmgV ...

  18. EST Table: FS834082 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/10 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fmgV ...

  19. EST Table: FS732056 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  20. EST Table: DC551466 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/02 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 phe- ...

  1. EST Table: FS734055 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  2. EST Table: FS771658 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/08 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fcaL ...

  3. EST Table: BY926430 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/29 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 F1mg ...

  4. EST Table: FS743784 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/07 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 bmmt ...

  5. EST Table: FS871886 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/11 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS834881 fner ...

  6. EST Table: FS730064 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/03 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 bmmt ...

  7. EST Table: BY918125 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/29 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h DN237205 F1mg ...

  8. EST Table: BY936252 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/30 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h DN237205 prW- ...

  9. EST Table: FS799456 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ina] gb|ADE18271.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18284.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18388.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18557.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18583.1| ...NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18661.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18713.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/09/09 n.h 10/08/29 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h BB990129 fmgV ...

  10. Structure of a bacterial enzyme regulated by phosphorylation, isocitrate dehydrogenase.

    OpenAIRE

    1989-01-01

    The structure of isocitrate dehydrogenase [threo-DS-isocitrate: NADP+ oxidoreductase (decarboxylating), EC 1.1.1.42] from Escherichia coli has been solved and refined at 2.5 A resolution and is topologically different from that of any other dehydrogenase. This enzyme, a dimer of identical 416-residue subunits, is inactivated by phosphorylation at Ser-113, which lies at the edge of an interdomain pocket that also contains many residues conserved between isocitrate dehydrogenase and isopropylma...

  11. Malate dehydrogenase: a model for structure, evolution, and catalysis.

    OpenAIRE

    1994-01-01

    Malate dehydrogenases are widely distributed and alignment of the amino acid sequences show that the enzyme has diverged into 2 main phylogenetic groups. Multiple amino acid sequence alignments of malate dehydrogenases also show that there is a low degree of primary structural similarity, apart from in several positions crucial for nucleotide binding, catalysis, and the subunit interface. The 3-dimensional structures of several malate dehydrogenases are similar, despite their low amino acid s...

  12. Kinetics of myoglobin redox form stabilization by malate dehydrogenase.

    Science.gov (United States)

    Mohan, Anand; Muthukrishnan, S; Hunt, Melvin C; Barstow, Thomas J; Houser, Terry A

    2010-06-09

    This study reports the reduction of metmyoglobin (MMb) via oxidation of malate to oxaloacetate and the regeneration of reduced nicotinamide adenine dinucleotide (NADH) via malate dehydrogenase (MDH). Two experiments were conducted to evaluate a malate-MDH-NADH system as a possible mechanism for MMb reduction. In experiment 1, kinetics of MDH and MMb reduction were determined, and the results showed that increasing concentrations of oxidized nicotinamide adenine dinucleotide (NAD(+)) and l-malate also increased (p malate and NAD(+) added. Reduction of MMb in the muscle extracts via MDH was NAD(+), malate, and extract concentration dependent (p malate can replenish NADH via MDH activity in post-mortem muscle, ultimately resulting in a more functional meat color.

  13. A highly efficient ADH-coupled NADH-recycling system for the asymmetric bioreduction of carbon-carbon double bonds using enoate reductases.

    Science.gov (United States)

    Tauber, Katharina; Hall, Melanie; Kroutil, Wolfgang; Fabian, Walter M F; Faber, Kurt; Glueck, Silvia M

    2011-06-01

    The asymmetric bioreduction of activated alkenes catalyzed by flavin-dependent enoate reductases from the OYE-family represents a powerful method for the production of optically active compounds. For its preparative-scale application, efficient and economic NADH-recycling is crucial. A novel enzyme-coupled NADH-recycling system is proposed based on the concurrent oxidation of a sacrificial sec-alcohol catalyzed by an alcohol dehydrogenase (ADH-A). Due to the highly favorable position of the equilibrium of ene-reduction versus alcohol-oxidation, the cosubstrate is only required in slight excess.

  14. Contribution of NADH increases to ethanol’s inhibition of retinol oxidation by human ADH isoforms

    Science.gov (United States)

    Chase, Jennifer R.; Poolman, Mark G.; Fell, David A.

    2010-01-01

    Background A decrease in retinoic acid levels due to alcohol consumption has been proposed as a contributor to such conditions as fetal alcohol spectrum diseases and ethanol-induced cancers. One molecular mechanism, competitive inhibition by ethanol of the catalytic activity of human alcohol dehydrogenase (EC 1.1.1.1) (ADH) on all-trans retinol oxidation has been shown for the ADH7 isoform. Ethanol metabolism also causes an increase in the free NADH in cells, which might reasonably be expected to decrease the retinol oxidation rate by product inhibition of ADH isoforms. Method To understand the relative importance of these two mechanisms by which ethanol decreases the retinol oxidation in vivo we need to assess them quantitatively. We have built a model system of four reactions: (1) ADH oxidation of ethanol and NAD+ (2) ADH oxidation of retinol and NAD+ (3) oxidation of ethanol by a generalized Ethanoloxidase that uses NAD+ (4) NADHoxidase which carries out NADH turnover. Results Using the metabolic modeling package SCRUMPY, we have shown that the ethanol-induced increase in NADH contributes from 0–90% of the inhibition by ethanol, depending on [ethanol] and ADH isoform. Furthermore, while the majority of flux control of retinaldehyde production is exerted by ADH, Ethanoloxidase and the NADHoxidase contribute as well. Discussion Our results show that the ethanol-induced increase in NADH makes a contribution of comparable importance to the ethanol competitive inhibition throughout the range of conditions likely to occur in vivo, and must be considered in the assessment of the in vivo mechanism of ethanol interference with fetal development and other diseases. PMID:19183134

  15. NADH/NADPH Oxidase and Vascular Function.

    Science.gov (United States)

    Griendling, K K; Ushio-Fukai, M

    1997-11-01

    The vascular NADH/NADPH oxidase has been shown to be the major source of superoxide in the vessel wall. Recent work has provided insight into its structure and activity in vascular cells. This enzyme is involved in both vascular smooth muscle hypertrophy and in some forms of impaired endothelium-dependent relaxation. Because oxidative stress in general participates in the pathogenesis of hypertension and atherosclerosis, the enzymes that produce reactive oxygen species may be important determinants of the course of vascular disease. (Trends Cardiovasc Med 1997;7:301-307). © 1997, Elsevier Science Inc.

  16. Purification of Mitochondrial Glutamate Dehydrogenase from Dark-Grown Soybean Seedlings.

    Science.gov (United States)

    Turano, F. J.; Dashner, R.; Upadhyaya, A.; Caldwell, C. R.

    1996-11-01

    Proteins in extracts from cotyledons, hypocotyls, and roots of 5-d-old, dark-grown soybean (Glycine max L. Merr. cv Williams) seedlings were separated by polyacrylamide gel electrophoresis. Three isoforms of glutamate dehydrogenase (GDH) were resolved and visualized in gels stained for GDH activity. Two isoforms with high electrophoretic mobility, GDH1 and GDH2, were in protein extracts from cotyledons and a third isoform with the lowest electrophoretic mobility, GDH3, was identified in protein extracts from root and hypocotyls. Subcellular fractionation of dark-grown soybean tissues demonstrated that GDH3 was associated with intact mitochondria. GDH3 was purified to homogeneity, as determined by native and sodium dodecyl sulfate-polyacrylamide gels. The isoenzyme was composed of a single 42-kD subunit. The pH optima for the reductive amination and the oxidative deamination reactions were 8.0 and 9.3, respectively. At any given pH, GDH activity was 12- to 50-fold higher in the direction of reductive amination than in the direction of the oxidative deamination reaction. GDH3 had a cofactor preference for NAD(H) over NADP(H). The apparent Michaelis constant values for [alpha]-ketoglutarate, ammonium, and NADH at pH 8.0 were 3.6, 35.5, and 0.07 mM, respectively. The apparent Michaelis constant values for glutamate and NAD were 15.8 and 0.10 mM at pH 9.3, respectively. To our knowledge, this is the first biochemical and physical characterization of a purified mitochondrial NAD(H)-dependent GDH isoenzyme from soybean.

  17. 2-Butanol and butanone production in Saccharomyces cerevisiae through combination of a B12 dependent dehydratase and a secondary alcohol dehydrogenase using a TEV-based expression system.

    Directory of Open Access Journals (Sweden)

    Payam Ghiaci

    Full Text Available 2-Butanol and its chemical precursor butanone (methyl ethyl ketone--MEK are chemicals with potential uses as biofuels and biocommodity chemicals. In order to produce 2-butanol, we have demonstrated the utility of using a TEV-protease based expression system to achieve equimolar expression of the individual subunits of the two protein complexes involved in the B12-dependent dehydratase step (from the pdu-operon of Lactobacillus reuteri, which catalyze the conversion of meso-2,3-butanediol to butanone. We have furthermore identified a NADH dependent secondary alcohol dehydrogenase (Sadh from Gordonia sp. able to catalyze the subsequent conversion of butanone to 2-butanol. A final concentration of 4±0.2 mg/L 2-butanol and 2±0.1 mg/L of butanone was found. A key factor for the production of 2-butanol was the availability of NADH, which was achieved by growing cells lacking the GPD1 and GPD2 isogenes under anaerobic conditions.

  18. Single-walled carbon nanotubes covalently functionalized with polytyrosine: A new material for the development of NADH-based biosensors.

    Science.gov (United States)

    Eguílaz, Marcos; Gutierrez, Fabiana; González-Domínguez, Jose Miguel; Martínez, María T; Rivas, Gustavo

    2016-12-15

    We report for the first time the use of single-walled carbon nanotubes (SWCNT) covalently functionalized with polytyrosine (Polytyr) (SWCNT-Polytyr) as a new electrode material for the development of nicotinamide adenine dinucleotide (NADH)-based biosensors. The oxidation of glassy carbon electrodes (GCE) modified with SWCNT-Polytyr at potentials high enough to oxidize the tyrosine residues have allowed the electrooxidation of NADH at low potentials due to the catalytic activity of the quinones generated from the primary oxidation of tyrosine without any additional redox mediator. The amperometric detection of NADH at 0.200V showed a sensitivity of (217±3)µAmM(-1)cm(-2) and a detection limit of 7.9nM. The excellent electrocatalytic activity of SWCNT-Polytyr towards NADH oxidation has also made possible the development of a sensitive ethanol biosensor through the immobilization of alcohol dehydrogenase (ADH) via Nafion entrapment, with excellent analytical characteristics (sensitivity of (5.8±0.1)µAmM(-1)cm(-2), detection limit of 0.67µM) and very successful application for the quantification of ethanol in different commercial beverages.

  19. Enzymatic reduction of complex redox dyes using NADH-dependent reductase from Bacillus subtilis coupled with cofactor regeneration.

    Science.gov (United States)

    Bozic, Mojca; Pricelius, Sina; Guebitz, Georg M; Kokol, Vanja

    2010-01-01

    Conventional vat dyeing involves chemical reduction of dyes into their water-soluble leuco form generating considerable amounts of toxic chemicals in effluents. In the present study, a new beta-nicotinamide adenine dinucleotide disodium salt (NADH)-dependent reductase isolated from Bacillus subtilis was used to reduce the redox dyes CI Acid Blue 74, CI Natural Orange 6, and CI Vat Blue 1 into their water-soluble leuco form. Enzymatic reduction was optimized in relation to pH and temperature conditions. The reductase was able to reduce Acid Blue 74 and Natural Orange 6 in the presence of the stoichiometrically consumed cofactor NADH; meanwhile, Vat Blue 1 required the presence of mediator 1,8-dihydroxyanthraquinone. Oxygen from air was used to reoxidize the dyes into their initial forms. The enzymatic reduction of the dyes was studied and the kinetic constants determined, and these were compared to the chemically-reduced leuco form. The enzyme responsible for the reduction showed homology to a NADH-dependent reductase from B. subtilis based on results from the MS/MS peptide mass mapping of the tryptically digested protein. Additionally, the reduction of Acid Blue 74 to its leuco form by reductase from B. subtilis was confirmed using NADH regenerated by the oxidation of formic acid with formate dehydrogenase from Candida boidinii in the same solution.

  20. Assembly of the Escherichia coli NADH:ubiquinone oxidoreductase (respiratory complex I).

    Science.gov (United States)

    Friedrich, Thorsten; Dekovic, Doris Kreuzer; Burschel, Sabrina

    2016-03-01

    Energy-converting NADH:ubiquinone oxidoreductase, respiratory complex I, couples the electron transfer from NADH to ubiquinone with the translocation of four protons across the membrane. The Escherichia coli complex I is made up of 13 different subunits encoded by the so-called nuo-genes. The electron transfer is catalyzed by nine cofactors, a flavin mononucleotide and eight iron-sulfur (Fe/S)-clusters. The individual subunits and the cofactors have to be assembled together in a coordinated way to guarantee the biogenesis of the active holoenzyme. Only little is known about the assembly of the bacterial complex compared to the mitochondrial one. Due to the presence of so many Fe/S-clusters the assembly of complex I is intimately connected with the systems responsible for the biogenesis of these clusters. In addition, a few other proteins have been reported to be required for an effective assembly of the complex in other bacteria. The proposed role of known bacterial assembly factors is discussed and the information from other bacterial species is used in this review to draw an as complete as possible model of bacterial complex I assembly. In addition, the supramolecular organization of the complex in E. coli is briefly described. This article is part of a Special Issue entitled Organization and dynamics of bioenergetic systems in bacteria, edited by Prof. Conrad Mullineaux.

  1. Localization and function of the membrane-bound riboflavin in the Na+-translocating NADH:quinone oxidoreductase (Na+-NQR) from Vibrio cholerae.

    Science.gov (United States)

    Casutt, Marco S; Huber, Tamara; Brunisholz, René; Tao, Minli; Fritz, Günter; Steuber, Julia

    2010-08-27

    The sodium ion-translocating NADH:quinone oxidoreductase (Na(+)-NQR) from the human pathogen Vibrio cholerae is a respiratory membrane protein complex that couples the oxidation of NADH to the transport of Na(+) across the bacterial membrane. The Na(+)-NQR comprises the six subunits NqrABCDEF, but the stoichiometry and arrangement of these subunits are unknown. Redox-active cofactors are FAD and a 2Fe-2S cluster on NqrF, covalently attached FMNs on NqrB and NqrC, and riboflavin and ubiquinone-8 with unknown localization in the complex. By analyzing the cofactor content and NADH oxidation activity of subcomplexes of the Na(+)-NQR lacking individual subunits, the riboflavin cofactor was unequivocally assigned to the membrane-bound NqrB subunit. Quantitative analysis of the N-terminal amino acids of the holo-complex revealed that NqrB is present in a single copy in the holo-complex. It is concluded that the hydrophobic NqrB harbors one riboflavin in addition to its covalently attached FMN. The catalytic role of two flavins in subunit NqrB during the reduction of ubiquinone to ubiquinol by the Na(+)-NQR is discussed.

  2. Role of lactate dehydrogenase in metmyoglobin reduction and color stability of different bovine muscles.

    Science.gov (United States)

    Kim, Y H; Keeton, J T; Smith, S B; Berghman, L R; Savell, J W

    2009-11-01

    The role of lactate dehydrogenase (LDH) in metmyoglobin reducing activity (MRA) and color stability of different bovine muscles was studied in two consecutive experiments. In experiment 1, three different bovine muscles -M. longissimus lumborum (LL), M. semimembranosus (SM), and M. psoas major (PM) - were obtained (n=7, respectively), cut into steaks, PVC packaged, and then displayed for 7days at 1°C. The LL was the most red over display time and had more (PLDH-B activity (catalyzing toward NADH generation), LDH1 isoform expression, NADH, and higher (PLDH-B activity, NADH, and a* values after 10days display at 1°C. These results suggest that variation in color stability of physiologically different muscles is regulated by different replenishment rates of NADH via different LDH isozymes.

  3. Purification, properties, and kinetic studies of cytoplasmic malate dehydrogenase from Taenia solium cysticerci.

    Science.gov (United States)

    Plancarte, Agustín; Nava, Gabriela; Mendoza-Hernández, Guillermo

    2009-07-01

    Malate dehydrogenase (L: -malate: NAD oxidoreductase, EC 1.1.1.37) from the cytoplasm of Taenia solium cysticerci (cMDHTs) was purified 48-fold through a four-step procedure involving salt fractionation, ionic exchange, and dye affinity chromatography. cMDHTs had a native M (r) of 64,000, while the corresponding value per subunit, obtained under denaturing conditions, was 32,000. The enzyme is partially positive, with an isoelectric point of 8.7, and had a specific activity of 2,615 U mg(-1) in the reduction of oxaloacetate. The second to the 21st amino acids from cMDHTs N-terminal group were P G P L R V L I T G A A G Q I A Y N L S. This sequence is 100% identical to that of Echinococcus granulosus. Basic kinetic parameters were determined for this enzyme. The optimum pH for enzyme reaction was at 7.6 for oxaloacetate reduction and at 9.6 for malate oxidation. K (m) values for oxaloacetate, malate, NAD, and NADH were 2.4, 215, 50, and 48 microM, respectively. V (max) values for the substrates and cosubstrates as described above were 1,490, 87.8, 104, and 1,714 micromol min(-1) mg(-1). Several NAD analogs, structurally altered in either the pyridine or purine moiety, were observed to function as coenzymes in the reaction catalyzed by the purified malate dehydrogenase. cMDHTs activity was uncompetitive inhibited by arsenate for both the forward (Ki = 8.2 mM) and reverse (Ki = 77 mM) reactions. The mechanism of the cMDHTs reactivity was investigated kinetically by the product inhibition approach. The results of this study are qualitatively consistent with an Ordered Bi Bi reaction mechanism, in which only the coenzymes can react with the free enzyme.

  4. Studies on the mechanism of electron bifurcation catalyzed by electron transferring flavoprotein (Etf) and butyryl-CoA dehydrogenase (Bcd) of Acidaminococcus fermentans.

    Science.gov (United States)

    Chowdhury, Nilanjan Pal; Mowafy, Amr M; Demmer, Julius K; Upadhyay, Vikrant; Koelzer, Sebastian; Jayamani, Elamparithi; Kahnt, Joerg; Hornung, Marco; Demmer, Ulrike; Ermler, Ulrich; Buckel, Wolfgang

    2014-02-21

    Electron bifurcation is a fundamental strategy of energy coupling originally discovered in the Q-cycle of many organisms. Recently a flavin-based electron bifurcation has been detected in anaerobes, first in clostridia and later in acetogens and methanogens. It enables anaerobic bacteria and archaea to reduce the low-potential [4Fe-4S] clusters of ferredoxin, which increases the efficiency of the substrate level and electron transport phosphorylations. Here we characterize the bifurcating electron transferring flavoprotein (EtfAf) and butyryl-CoA dehydrogenase (BcdAf) of Acidaminococcus fermentans, which couple the exergonic reduction of crotonyl-CoA to butyryl-CoA to the endergonic reduction of ferredoxin both with NADH. EtfAf contains one FAD (α-FAD) in subunit α and a second FAD (β-FAD) in subunit β. The distance between the two isoalloxazine rings is 18 Å. The EtfAf-NAD(+) complex structure revealed β-FAD as acceptor of the hydride of NADH. The formed β-FADH(-) is considered as the bifurcating electron donor. As a result of a domain movement, α-FAD is able to approach β-FADH(-) by about 4 Å and to take up one electron yielding a stable anionic semiquinone, α-FAD, which donates this electron further to Dh-FAD of BcdAf after a second domain movement. The remaining non-stabilized neutral semiquinone, β-FADH(•), immediately reduces ferredoxin. Repetition of this process affords a second reduced ferredoxin and Dh-FADH(-) that converts crotonyl-CoA to butyryl-CoA.

  5. Measurement of mitochondrial NADH and FAD autofluorescence in live cells.

    Science.gov (United States)

    Bartolomé, Fernando; Abramov, Andrey Y

    2015-01-01

    In the process of energy production, mitochondrial networks are key elements to allow metabolism of substrates into ATP. Many pathological conditions have been associated with mitochondrial dysfunction as mitochondria are associated with a wide range of cellular processes. Therefore, any disruption in the energy production induces devastating effects that can ultimately lead to cell death due to chemical ischemia. To address the mitochondrial health and function, there are several bioenergetic parameters reflecting either whole mitochondrial functionality or individual mitochondrial complexes. Particularly, metabolism of nutrients in the tricarboxylic acid cycle provides substrates used to generate electron carriers (nicotinamide adenine dinucleotide [NADH] and flavin adenine dinucleotide [FADH2]) which ultimately donate electrons to the mitochondrial electron transport chain. The levels of NADH and FADH2 can be estimated through imaging of NADH/NAD(P)H or FAD autofluorescence. This report demonstrates how to perform and analyze NADH/NAD(P)H and FAD autofluorescence in a time-course-dependent manner and provides information about NADH and FAD redox indexes both reflecting the activity of the mitochondrial electron transport chain (ETC). Furthermore, total pools of NADH and FAD can be estimated providing information about the rate of substrate supply into the ETC. Finally, the analysis of NADH autofluorescence after induction of maximal respiration can offer information about the pentose phosphate pathway activity where glucose can be alternatively oxidized instead of pyruvate.

  6. High performance enzyme fuel cells using a genetically expressed FAD-dependent glucose dehydrogenase α-subunit of Burkholderia cepacia immobilized in a carbon nanotube electrode for low glucose conditions.

    Science.gov (United States)

    Fapyane, Deby; Lee, Soo-Jin; Kang, Seo-Hee; Lim, Du-Hyun; Cho, Kwon-Koo; Nam, Tae-hyun; Ahn, Jae-Pyoung; Ahn, Jou-Hyeon; Kim, Seon-Won; Chang, In Seop

    2013-06-28

    FAD-dependent glucose dehydrogenase (FAD-GDH) of Burkholderia cepacia was successfully expressed in Escherichia coli and subsequently purified in order to use it as an anode catalyst for enzyme fuel cells. The purified enzyme has a low Km value (high affinity) towards glucose, which is 463.8 μM, up to 2-fold exponential range lower compared to glucose oxidase. The heterogeneous electron transfer coefficient (Ks) of FAD-GDH-menadione on a glassy carbon electrode was 10.73 s(-1), which is 3-fold higher than that of GOX-menadione, 3.68 s(-1). FAD-GDH was able to maintain its native glucose affinity during immobilization in the carbon nanotube and operation of enzyme fuel cells. FAD-GDH-menadione showed 3-fold higher power density, 799.4 ± 51.44 μW cm(-2), than the GOX-menadione system, 308.03 ± 17.93 μW cm(-2), under low glucose concentration, 5 mM, which is the concentration in normal physiological fluid.

  7. An NAD-specific glutamate dehydrogenase from cyanobacteria. Identification and properties.

    Science.gov (United States)

    Chávez, S; Candau, P

    1991-07-08

    The unicellular cyanobacterium Synechocystis sp. PCC 6803 presents a hexameric NAD-specific glutamate dehydrogenase with a molecular mass of 295 kDa. The enzyme differs from the NADP-glutamate dehydrogenase found in the same strain and is coded by a different gene. NAD-glutamate dehydrogenase shows a high coenzyme specificity, catalyzes preferentially glutamate formation and presents Km values for ammonium, NADH and 2-oxoglutarate of 4.5 mM, 50 microM and 1.8 mM respectively. An animating role for the enzyme is discussed.

  8. 小分子干扰RNA抑制高氧暴露下人肺腺癌A549细胞中的硫氧还蛋白-2对还原型烟酰胺腺嘌呤二核苷酸脱氢酶亚单位1、细胞色素C氧化酶工表达的影响%Suppressed expression of thioredoxin-2 by small interference RNA in A549 cells exposed to hyperoxia reduced expression of nicotinamide-adenine dinucleotide dehydrogenase subunit 1 and cytochrome C oxidase Ⅰ

    Institute of Scientific and Technical Information of China (English)

    蔡成; 常立文; 李文斌; 陈燕; 单瑞艳; 刘伟; 潘睿

    2010-01-01

    Objective To explore the effects of expression of thioredoxin-2(Trx-2) suppressed by small interference RNA(SiRNA) in A549 cells exposed to hyperoxia on expression of nicotinamide adenine dinucleotide(NADH) dehydrogenase subunit 1(ND1)and cytochrome C oxidase Ⅰ(COX Ⅰ). Methods A549 cells were gained by serial subcultivation in vitro and transfered with synthetic Trx-2 sequence-specific SiRNA and then were randomly divided into air group without interference,hyperoxia group without interference,air group after interference,and hyperoxia group after interference.After exposure to oxygen or room air for 12,24 and 48 h,expressions of Trx-2,ND1 and COX Ⅰ mRNA of these cells were detected by reverse transcription-polymerase chain reaction (RT-PCR),and Trx-2 protein was detected by Western blot. Results (1)Sequence-specific SiRNA targeting Trx-2 could significantly down-regulate its expression in A549 cells.(2)Trx-2 mRNA levds in hyperoxia group without interference at 24 h was higher than those in air group without interference(0.7799±0.1249 VS 0.4424±Ⅰ.1140,P<0.05).Th-2 mRNA levels in hyperoxia group after ireedcrence at 24 h and 48 h were 0.2774±0.0174 and 0.2587±0.0069,lower than those in air group after interference and hyperoxia group without interference (P<0.05).(3)ND1 mRNA levels in hyperoxia group without interference at 24 h was 0.6609±0.0368,lower than those in air group without interference(0.8898±0.1049)(P<0.05).ND1 mRNA levels in hyperoxia group after interference at 12 h was 0.8848±0.0135,higher than those in air group after imederence(P<0.05).ND1 mRNA levels in hypemxia group after interference at 48 h was 0.3808±0.0937,lower than those in air group after imerference and hyperoxia group without interference(P<0.05).(4)COXI mRNA levels in hypemxia group without inteference at 12,24 and 48 h were 1.7313±0.4331,2.1929±0.6722 and 2.0754±0.2584,higher than those in air group witheUt interference,respectively (P<0.05). Conclusions ND1 and

  9. The Role of Pyruvate Dehydrogenase Kinase in Diabetes and Obesity

    Directory of Open Access Journals (Sweden)

    In-Kyu Lee

    2014-06-01

    Full Text Available The pyruvate dehydrogenase complex (PDC is an emerging target for the treatment of metabolic syndrome. To maintain a steady-state concentration of adenosine triphosphate during the feed-fast cycle, cells require efficient utilization of fatty acid and glucose, which is controlled by the PDC. The PDC converts pyruvate, coenzyme A (CoA, and oxidized nicotinamide adenine dinucleotide (NAD+ into acetyl-CoA, reduced form of nicotinamide adenine dinucleotide (NADH, and carbon dioxide. The activity of the PDC is up- and down-regulated by pyruvate dehydrogenase kinase and pyruvate dehydrogenase phosphatase, respectively. In addition, pyruvate is a key intermediate of glucose oxidation and an important precursor for the synthesis of glucose, glycerol, fatty acids, and nonessential amino acids.

  10. Glucose-6-phosphate dehydrogenase

    Science.gov (United States)

    ... medlineplus.gov/ency/article/003671.htm Glucose-6-phosphate dehydrogenase test To use the sharing features on this page, please enable JavaScript. Glucose-6-phosphate dehydrogenase (G6PD) is a protein that helps red ...

  11. Lactate dehydrogenase test

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003471.htm Lactate dehydrogenase test To use the sharing features on this page, please enable JavaScript. Lactate dehydrogenase (LDH) is a protein that helps produce energy ...

  12. Mitochondrial dynamics in human NADH:ubiquinone oxidoreductase deficiency.

    NARCIS (Netherlands)

    Willems, P.H.G.M.; Smeitink, J.A.M.; Koopman, W.J.H.

    2009-01-01

    Mitochondrial NADH:ubiquinone oxidoreductase or complex I (CI) is a frequently affected enzyme in cases of mitochondrial disorders. However, the cytopathological mechanism of the associated pediatric syndromes is poorly understood. Evidence in the literature suggests a connection between mitochondri

  13. Hybridizability of gamma-irradiated lactic dehydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Saito, M.

    1976-03-01

    The hybridizabilities of the gamma-irradiated chicken heart and pig muscle lactic dehydrogenases were estimated by hybridizing the irradiated enzymes with the unirradiated pig heart lactic dehydrogenase. The disc gel electrophoretic patterns of the inter- and intraspecific hybrids showed that the LDH activity of the pig heart isozyme band increased as a function of dose. This observation was analyzed upon the binomial redistribution pattern of the recombined subunits. The result shows that the hybridizabilities of both the chicken heart and pig muscle isozymes decreased along with the loss of catalytic activity and the release from substrate inhibition. The titration of free SH groups of the irradiated chicken isozyme suggested that the unfolding of the peptide chain destroyed the specific tertiary structure needed for the binding of subunits. (auth)

  14. A novel glutamate dehydrogenase from bovine brain: purification and characterization.

    Science.gov (United States)

    Lee, J; Kim, S W; Cho, S W

    1995-08-01

    A soluble form of novel glutamate dehydrogenase has been purified from bovine brain. The preparation was homogeneous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and composed of six identical subunits having a subunit size of 57,500 Da. The biochemical properties of glutamate dehydrogenase such as N-terminal amino acids sequences, kinetic parameters, amino acids analysis, and optimum pH were examined in both reductive amination of alpha-ketoglutarate and oxidative deamination of glutamate. N-terminal amino acid sequences of the bovine brain enzyme showed the significant differences in the first 5 amino acids compared to other glutamate dehydrogenases from various sources. These results indicate that glutamate dehydrogenase isolated from bovine brain is a novel polypeptide.

  15. A New Biochemical Way for Conversion of CO2 to Methanol via Dehydrogenases Encapsulated in SiO2 Matrix

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    CO2 is converted to methanol through an enzymatic approach using formate dehydro- genase (FateDH), formaldehyde dehydrogenase (FaldDH) and alcohol dehydrogenase (ADH) co- encapsulated in silica gel prepared by modified sol-gel process as catalysts, TEOS as precursor, NADH as an electron donor. The highest yield of methanol was up to 92.1% under 37℃, pH7.0 and 0.3Mpa.

  16. Mitochondrial Ca2+ homeostasis in human NADH:ubiquinone oxidoreductase deficiency.

    Science.gov (United States)

    Willems, Peter H G M; Valsecchi, Federica; Distelmaier, Felix; Verkaart, Sjoerd; Visch, Henk-Jan; Smeitink, Jan A M; Koopman, Werner J H

    2008-07-01

    NADH:ubiquinone oxidoreductase or complex I is a large multisubunit assembly of the mitochondrial inner membrane that channels high-energy electrons from metabolic NADH into the electron transport chain (ETC). Its dysfunction is associated with a range of progressive neurological disorders, often characterized by a very early onset and short devastating course. To better understand the cytopathological mechanisms of these disorders, we use live cell luminometry and imaging microscopy of patient skin fibroblasts with mutations in nuclear-encoded subunits of the complex. Here, we present an overview of our recent work, showing that mitochondrial membrane potential, Ca(2+) handling and ATP production are to a variable extent impaired among a large cohort of patient fibroblast lines. From the results obtained, the picture emerges that a reduction in cellular complex I activity leads to a depolarization of the mitochondrial membrane potential, resulting in a decreased supply of mitochondrial ATP to the Ca(2+)-ATPases of the intracellular stores and thus to a reduced Ca(2+) content of these stores. As a consequence, the increase in cytosolic Ca(2+) concentration evoked by a Ca(2+) mobilizing stimulus is decreased, leading to a reduction in mitochondrial Ca(2+) accumulation and ensuing ATP production and thus to a hampered energization of stimulus-induced cytosolic processes.

  17. Isolation, characterization and evaluation of the Pichia pastoris sorbitol dehydrogenase promoter for expression of heterologous proteins.

    Science.gov (United States)

    Periyasamy, Sankar; Govindappa, Nagaraj; Sreenivas, Suma; Sastry, Kedarnath

    2013-11-01

    Sorbitol is used as a non-repressive carbon source to develop fermentation process for Mut(s) recombinant clones obtained using the AOX1 promoter in Pichia pastoris. Sorbitol dehydrogenase is an enzyme in the carbohydrate metabolism that catalyzes reduction of D-fructose into D-sorbitol in the presence of NADH. The small stretch of 211bps upstream region of sorbitol dehydrogenase coding gene has all the promoter elements like CAAT box, GC box, etc. It is able to promote protein production under repressive as well as non-repressive carbon sources. In this study, the strength of the sorbitol dehydrogenase promoter was evaluated by expression of two heterologous proteins: human serum albumin and erythrina trypsin inhibitor. Sorbitol dehydrogenase promoter allowed constitutive expression of recombinant proteins in all carbon sources that were tested to grow P. pastoris and showed activity similar to GAP promoter. The sorbitol dehydrogenase promoter was active in all the growth phases of the P. pastoris.

  18. The free NADH concentration is kept constant in plant mitochondria under different metabolic conditions

    DEFF Research Database (Denmark)

    Kasimova, M.R.; Grigiene, J.; Krab, K.

    2006-01-01

    The reduced coenzyme NADH plays a central role in mitochondrial respiratory metabolism. However, reports on the amount of free NADH in mitochondria are sparse and contradictory. We first determined the emission spectrum of NADH bound to proteins using isothermal titration calorimetry combined...... with fluorescence spectroscopy. The NADH content of actively respiring mitochondria (from potato tubers [Solanum tuberosum cv Bintje]) in different metabolic states was then measured by spectral decomposition analysis of fluorescence emission spectra. Most of the mitochondrial NADH is bound to proteins...

  19. Probing the proton channels in subunit N of Complex I from Escherichia coli through intra-subunit cross-linking.

    Science.gov (United States)

    Tursun, Ablat; Zhu, Shaotong; Vik, Steven B

    2016-12-01

    Respiratory Complex I appears to have 4 sites for proton translocation, which are coupled to the oxidation of NADH and reduction of coenzyme Q. The proton pathways are thought to be made of offset half-channels that connect to the membrane surfaces, and are connected by a horizontal path through the center of the membrane. In this study of the enzyme from Escherichia coli, subunit N, containing one of the sites, was targeted. Pairs of cysteine residues were introduced into neighboring α-helices along the proposed proton pathways. In an effort to constrain conformational changes that might occur during proton translocation, we attempted to form disulfide bonds or methanethiosulfonate bridges between two engineered cysteine residues. Cysteine modification was inferred by the inability of PEG-maleimide to shift the electrophoretic mobility of subunit N, which will occur upon reaction with free sulfhydryl groups. After the cross-linking treatment, NADH oxidase and NADH-driven proton translocation were measured. Ten different pairs of cysteine residues showed evidence of cross-linking. The most significant loss of enzyme activity was seen for residues near the essential Lys 395. This residue is positioned between the proposed proton half-channel to the periplasm and the horizontal connection through subunit N, and is also near the essential Glu 144 of subunit M. The results suggest important conformational changes in this region for the delivery of protons to the periplasm, or for coupling the actions of subunit N to subunit M. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. NCBI nr-aa BLAST: CBRC-CFAM-25-0003 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CFAM-25-0003 gb|AAP74617.1|AF484337_1 NADH dehydrogenase subunit 4 [Venerupis (Ruditapes) philip...pinarum] gb|AAP74624.1|AF484338_1 NADH dehydrogenase subunit 4 [Venerupis (Ruditapes) philip...pinarum] gb|AAP74638.1|AF484340_1 NADH dehydrogenase subunit 4 [Venerupis (Ruditapes) philippinarum] AAP74617.1 5.1 30% ...

  1. NCBI nr-aa BLAST: CBRC-PCAP-01-0207 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PCAP-01-0207 gb|AAP74585.1|AF484332_8 NADH dehydrogenase subunit 5 [Venerupis philip...pinarum] gb|AAP74592.1|AF484333_13 NADH dehydrogenase subunit 5 [Venerupis philippinarum] gb|AAP74613....1|AF484336_18 NADH dehydrogenase subunit 5 [Venerupis philippinarum] AAP74585.1 0.72 29% ...

  2. AcEST: BP915326 [AcEST

    Lifescience Database Archive (English)

    Full Text Available 9ACAR NADH dehydrogenase subunit 4 OS=Unionicola foili Align length 54 Score (bit...W617_9ACAR NADH dehydrogenase subunit 4 OS=Unionicol... 33 9.2 tr|A7E3I5|A7E3I5_B...OMMO Odorant receptor 44 (Fragment) OS=Bombyx ... 33 9.2 >tr|B3W617|B3W617_9ACAR NADH dehydrogenase subunit 4 OS=Union

  3. The progress of pyruvate dehydrogenase E1 alpha subunit in myocardial ischemia-reperfusion injury%丙酮酸脱氢酶E1α亚单位与心肌缺血再灌注损伤的研究进展

    Institute of Scientific and Technical Information of China (English)

    叶星华; 梁贵友

    2016-01-01

    心肌缺血再灌注损伤(MIRI)是临床体外循环(CPB)心脏直视手术术后心功能障碍甚至导致死亡的主要原因之一,其发生机制至今仍未完全阐明.我们的前期研究结果显示,心肌胰岛素抵抗(IR)可能是MIRI的又一重要机制,涉及心肌能量底物葡萄糖和脂肪酸代谢紊乱.近来的研究结果显示,丙酮酸脱氢酶E1α亚单位(PDHA1)作为丙酮酸脱氢酶复合物(PDC)的重要组成部分,在维持缺血缺氧及再灌注心肌细胞糖、脂及能量代谢稳态中扮演关键的角色.通过研究PDHA1的相关分子机制,可进一步阐明体外循环心肌胰岛素抵抗的发生机制,对MIRI的防治具有重要意义.%The myocardial ischemia-reperfusion injury (MIRI) is one of the main reason to cardiac dysfunction which is even leading to the death after a open heart surgery by cardiopulmonary bypass (CPB).However,the mechanism of MIRI remains to be fully elucidated.Our previous studies have shown that myocardial insulin resistance (IR) might be another important mechanism of MIRI,involving myocardial energy substrate glucose and fatty acid metabolism disorders.Recent literatures indicated that pyruvate dehydrogenase E1 component subunit alpha (PDHA1)as the important part of pyruvate dehydrogenase complex (PDC),plays a key role in the maintenance of homeostasis of the carbohydrate,lipid and energy metabolism of ischemia and reperfusion myocardial cells.Through the study on molecular mechanisms of PDHA1,we can further elucidate the mechanism of CPB-myocardial IR and provide an important academic and practical significance forprevention and treatment of MIRI.

  4. Quinohemoprotein alcohol dehydrogenases: structure, function, and physiology.

    Science.gov (United States)

    Toyama, Hirohide; Mathews, F Scott; Adachi, Osao; Matsushita, Kazunobu

    2004-08-01

    Quino(hemo)protein alcohol dehydrogenases (ADH) that have pyrroloquinoline quinone (PQQ) as the prosthetic group are classified into 3 groups, types I, II, and III. Type I ADH is a simple quinoprotein having PQQ as the only prosthetic group, while type II and type III ADHs are quinohemoprotein having heme c as well as PQQ in the catalytic polypeptide. Type II ADH is a soluble periplasmic enzyme and is widely distributed in Proteobacteria such as Pseudomonas, Ralstonia, Comamonas, etc. In contrast, type III ADH is a membrane-bound enzyme working on the periplasmic surface solely in acetic acid bacteria. It consists of three subunits that comprise a quinohemoprotein catalytic subunit, a triheme cytochrome c subunit, and a third subunit of unknown function. The catalytic subunits of all the quino(hemo)protein ADHs have a common structural motif, a quinoprotein-specific superbarrel domain, where PQQ is deeply embedded in the center. In addition, in the type II and type III ADHs this subunit contains a unique heme c domain. Various type II ADHs each have a unique substrate specificity, accepting a wide variety of alcohols, as is discussed on the basis of recent X-ray crystallographic analyses. Electron transfer within both type II and III ADHs is discussed in terms of the intramolecular reaction from PQQ to heme c and also from heme to heme, and in terms of the intermolecular reaction with azurin and ubiquinone, respectively. Unique physiological functions of both types of quinohemoprotein ADHs are also discussed.

  5. NCBI nr-aa BLAST: CBRC-BTAU-01-2284 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-BTAU-01-2284 ref|NP_062836.1| NADH dehydrogenase subunit 6 [Loligo bleekeri] s...DH dehydrogenase 6 [Loligo bleekeri] dbj|BAB03648.1| NADH dehydrogenase subunit 6 [Loligo bleekeri] NP_062836.1 0.11 35% ...

  6. Assembly of respiratory complexes I, III, and IV into NADH oxidase supercomplex stabilizes complex I in Paracoccus denitrificans.

    Science.gov (United States)

    Stroh, Anke; Anderka, Oliver; Pfeiffer, Kathy; Yagi, Takao; Finel, Moshe; Ludwig, Bernd; Schägger, Hermann

    2004-02-01

    Stable supercomplexes of bacterial respiratory chain complexes III (ubiquinol:cytochrome c oxidoreductase) and IV (cytochrome c oxidase) have been isolated as early as 1985 (Berry, E. A., and Trumpower, B. L. (1985) J. Biol. Chem. 260, 2458-2467). However, these assemblies did not comprise complex I (NADH:ubiquinone oxidoreductase). Using the mild detergent digitonin for solubilization of Paracoccus denitrificans membranes we could isolate NADH oxidase, assembled from complexes I, III, and IV in a 1:4:4 stoichiometry. This is the first chromatographic isolation of a complete "respirasome." Inactivation of the gene for tightly bound cytochrome c552 did not prevent formation of this supercomplex, indicating that this electron carrier protein is not essential for structurally linking complexes III and IV. Complex I activity was also found in the membranes of mutant strains lacking complexes III or IV. However, no assembled complex I but only dissociated subunits were observed following the same protocols used for electrophoretic separation or chromatographic isolation of the supercomplex from the wild-type strain. This indicates that the P. denitrificans complex I is stabilized by assembly into the NADH oxidase supercomplex. In addition to substrate channeling, structural stabilization of a membrane protein complex thus appears as one of the major functions of respiratory chain supercomplexes.

  7. Influence of Altered NADH Metabolic Pathway on the Respiratory-deficient Mutant of Rhizopus oryzae and its L-lactate Production.

    Science.gov (United States)

    Shu, Chang; Guo, Chenchen; Luo, Shuizhong; Jiang, Shaotong; Zheng, Zhi

    2015-08-01

    Respiratory-deficient mutants of Rhizopus oryzae (R. oryzae) AS 3.3461 were acquired by ultraviolet (UV) irradiation to investigate changes in intracellular NADH metabolic pathway and its influence on the fermentation characteristics of the strain. Compared with R. oryzae AS 3.3461, the intracellular ATP level of the respiratory-deficient strain UV-1 decreased by 52.7 % and the glucose utilization rate rose by 8.9 %; When incubated for 36 h, the activities of phosphofructokinase (PFK), hexokinase (HK), and pyruvate kinase (PK) in the mutant rose by 74.2, 7.2, and 12.0 %, respectively; when incubated for 48 h, the intracellular NADH/NAD(+) ratio of the mutant rose by 14.6 %; when a mixed carbon source with a glucose/gluconic acid ratio of 1:1 was substituted to culture the mutant, the NADH/NAD(+) ratio decreased by 4.6 %; the ATP content dropped by 27.6 %; the lactate dehydrogenase (LDH) activity rose by 22.7 %; and the lactate yield rose by 11.6 %. These results indicated that changes to the NADH metabolic pathway under a low-energy charge level can effectively increase the glycolytic rate and further improve the yield of L-lactate of R. oryzae.

  8. Increasing NADH oxidation reduces overflow metabolism in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Vemuri, Goutham; Eiteman, M.A; McEwen, J.E

    2007-01-01

    Respiratory metabolism plays an important role in energy production in the form of ATP in all aerobically growing cells. However, a limitation in respiratory capacity results in overflow metabolism, leading to the formation of byproducts, a phenomenon known as ‘‘overflow metabolism’’ or ‘‘the...... Crabtree effect.’’ The yeast Saccharomyces cerevisiae has served as an important model organism for studying the Crabtree effect. When subjected to increasing glycolytic fluxes under aerobic conditions, there is a threshold value of the glucose uptake rate at which the metabolism shifts from purely...... by overexpression of a water-forming NADH oxidase reduced aerobic glycerol formation. The metabolic response to elevated alternative oxidase occurred predominantly in the mitochondria, whereas NADH oxidase affected genes that catalyze cytosolic reactions. Moreover, NADH oxidase restored the deficiency of cytosolic...

  9. EST Table: BY916369 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BY916369 mg0235 10/09/28 63 %/114 aa gb|ADE18193.1| NADH dehydrogenase subunit 6 [Bombyx mandarin...a] gb|ADE18206.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] gb|ADE18518.1| NADH dehydrog...enase subunit 6 [Bombyx mandarina] gb|ADE18687.1| NADH dehydrogenase subunit 6 [Bombyx mandarina] 10/08/29 n.h 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h DN237205 mg ...

  10. Identification of NADH kinase activity in filamentous fungi and structural modelling of the novel enzyme from Fusarium oxysporum

    DEFF Research Database (Denmark)

    Panagiotou, Gianni; Papadakis, Emmanouil; Topakas, E.

    2008-01-01

    ATP-NADH kinase phosphorylates NADH to produce NADPH at the expense of ATP. The present study describes Fusarium oxysporum NADH kinase (ATP:NADH 2'-phosphotransferase, EC 2.7.1.86), a novel fungal enzyme capable of synthesizing NADPH using NADH as the preferred diphosphonicotinamide (diphosphopyr...

  11. Purification and characterization of xylitol dehydrogenase from Fusarium oxysporum

    DEFF Research Database (Denmark)

    Panagiotou, Gianni; Kekos, D.; Macris, B.J.

    2002-01-01

    An NAD(+)-dependent xylitol dehydrogenase (XDH) from Fusarium oxysporum, a key enzyme in the conversion of xylose to ethanol, was purified to homogeneity and characterised. It was homodimeric with a subunit of M-r 48 000, and pI 3.6. It was optimally active at 45degreesC and pH 9-10. It was fully...

  12. Mutations associated with succinate dehydrogenase D-related malignant paragangliomas.

    NARCIS (Netherlands)

    Timmers, H.J.L.M.; Pacak, K.; Bertherat, J.; Lenders, J.W.M.; Duet, M.; Eisenhofer, G.; Stratakis, C.A.; Niccoli-Sire, P.; Tran, B.H.; Burnichon, N.; Gimenez-Roqueplo, A.P.

    2008-01-01

    OBJECTIVE: Hereditary paraganglioma (PGL) syndromes result from germline mutations in genes encoding subunits B, C and D of the mitochondrial enzyme succinate dehydrogenase (SDHB, SDHC and SDHD). SDHB-related PGLs are known in particular for their high malignant potential. Recently, however, maligna

  13. Biochemical characterization of ethanol-dependent reduction of furfural by alcohol dehydrogenases.

    Science.gov (United States)

    Li, Qunrui; Metthew Lam, L K; Xun, Luying

    2011-11-01

    Lignocellulosic biomass is usually converted to hydrolysates, which consist of sugars and sugar derivatives, such as furfural. Before yeast ferments sugars to ethanol, it reduces toxic furfural to non-inhibitory furfuryl alcohol in a prolonged lag phase. Bioreduction of furfural may shorten the lag phase. Cupriavidus necator JMP134 rapidly reduces furfural with a Zn-dependent alcohol dehydrogenase (FurX) at the expense of ethanol (Li et al. 2011). The mechanism of the ethanol-dependent reduction of furfural by FurX and three homologous alcohol dehydrogenases was investigated. The reduction consisted of two individual reactions: ethanol-dependent reduction of NAD(+) to NADH and then NADH-dependent reduction of furfural to furfuryl alcohol. The kinetic parameters of the coupled reaction and the individual reactions were determined for the four enzymes. The data indicated that limited NADH was released in the coupled reaction. The enzymes had high affinities for NADH (e.g., K ( d ) of 0.043 μM for the FurX-NADH complex) and relatively low affinities for NAD(+) (e.g., K ( d ) of 87 μM for FurX-NAD(+)). The kinetic data suggest that the four enzymes are efficient "furfural reductases" with either ethanol or NADH as the reducing power. The standard free energy change (ΔG°') for ethanol-dependent reduction of furfural was determined to be -1.1 kJ mol(-1). The physiological benefit for ethanol-dependent reduction of furfural is likely to replace toxic and recalcitrant furfural with less toxic and more biodegradable acetaldehyde.

  14. UniProt search blastx result: AK287459 [KOME

    Lifescience Database Archive (English)

    Full Text Available oroplast (EC 1.6.5.-) (NAD(P)H dehydrogenase subunit I) (NDH subunit I) (NADH-plastoquinone oxidoreductase subunit I) - Huperzia lucidula (Shining clubmoss) (Lycopodium lucidulum) 0 ...

  15. Differential Role of Glutamate Dehydrogenase in Nitrogen Metabolism of Maize Tissues 1

    Science.gov (United States)

    Loyola-Vargas, Victor Manuel; de Jimenez, Estela Sanchez

    1984-01-01

    Both calli and plantlets of maize (Zea mays L. var Tuxpeño 1) were exposed to specific nitrogen sources, and the aminative (NADH) and deaminative (NAD+) glutamate dehydrogenase activities were measured at various periods of time in homogenates of calli, roots, and leaves. A differential effect of the nitrogen sources on the tissues tested was observed. In callus tissue, glutamate, ammonium, and urea inhibited glutamate dehydrogenase (GDH) activity. The amination and deamination reactions also showed different ratios of activity under different nitrogen sources. In roots, ammonium and glutamine produced an increase in GDH-NADH activity whereas the same metabolites were inhibitory of this activity in leaves. These data suggest the presence of isoenzymes or conformers of GDH, specific for each tissue, whose activities vary depending on the nutritional requirements of the tissue and the state of differentiation. PMID:16663876

  16. Detection of ATP and NADH: A Bioluminescent Experience.

    Science.gov (United States)

    Selig, Ted C.; And Others

    1984-01-01

    Described is a bioluminescent assay for adenosine triphosphate (ATP) and reduced nicotineamide-adenine dinucleotide (NADH) that meets the requirements of an undergraduate biochemistry laboratory course. The 3-hour experiment provides students with experience in bioluminescence and analytical biochemistry yet requires limited instrumentation,…

  17. Human NADH : ubiquinone oxidoreductase deficiency : radical changes in mitochondrial morphology?

    NARCIS (Netherlands)

    Koopman, W.J.H.; Verkaart, S.A.J.; Visch, H.J.; Vries, S. de; Nijtmans, L.G.J.; Smeitink, J.A.M.; Willems, P.H.G.M.

    2007-01-01

    Malfunction of NADH:ubiquinone oxidoreductase or complex I (CI), the first and largest complex of the mitochondrial oxidative phosphorylation system, has been implicated in a wide variety of human disorders. To demonstrate a quantitative relationship between CI amount and activity and mitochondrial

  18. Studies on NADH (NADPH)-cytochrome c reductase (FMN-containing) from yeast. Isolation and physicochemical properties of the enzyme from top-fermenting ale yeast.

    Science.gov (United States)

    Johnson, M S; Kuby, S A

    1985-10-05

    Only three major NADPH-nitrotetrazolium blue (NTB) reductases may be detected in a unique top-ale yeast (Saccharomyces cerevisiae, Narragansett strain), which appears to be of a near anaerobic type with the absence of cytochromes c and a/a3 and the presence of cytochromes P-450 and b5. Two of these three major NADPH-NTB reductases possessed NADH-NTB reductase activity; the third was specific for NADPH and was isolated in this laboratory (Tryon, E., Cress, M. C., Hamada, M., and Kuby, S. A. (1979) Arch. Biochem. Biophys. 197, 104-118) vis. NADPH-cytochrome c reductase (FAD-containing). A description of the isolation procedure is provided for one of these two NADH(NADPH)-NTB reductases, viz. NADH(NADPH)-cytochrome c reductase (FMN-containing), which accounts for about one-half of the total cyanide-insensitive menadione-activated respiration of this yeast. This NADH(NADPH)-cytochrome c reductase has been isolated from an extract of an acetone powder of the top-fermenting ale yeast, with an apparent purification of more than 67-fold and a final specific activity of 0.41 and 0.31 mumol/min/mg for NADH- and NADPH-dependent reduction, respectively. The isolated enzyme proved to be homogeneous by electrophoresis on cellulose acetate and on polyacrylamide gels. It had a pI of 5.25 (at gamma/2 = 0.05) and a molecular size under nondenaturing conditions (as determined by chromatography on Sephadex G-100 and Sephacryl S-200) of 70,000 daltons. On denaturation, the enzyme dissociated into two similar, if not identical, subunits which possessed a molecular weight of 34,000 by sodium dodecyl sulfate/urea-polyacrylamide gel electrophoresis and a weight average molecular weight of 35,000 by sedimentation equilibrium in the presence of 4.0 M guanidinium chloride. The absorbance spectrum of NADH(NADPH)-cytochrome c reductase (FMN-containing) showed three maxima at 464, 383, and 278 nm, with extinction coefficients of 9.88, 9.98, and 64.6 mM-1 cm-1, respectively. The reductase, as

  19. Heterozygosity of the sheep: Polymorphism of 'malic enzyme', isocitrate dehydrogenase (NADP+), catalase and esterase.

    Science.gov (United States)

    Baker, C M; Manwell, C

    1977-04-01

    In contrast to other reports, it is found that the sheep has approximately as much enzyme variation as man. Most of the genetically interpretable enzyme variation in heart, liver, kidney and muscle from 52 sheep (Merinos or Merino crosses) is in the NADP-dependent dehydrogenases [two 'malic enzymes' and the supernatant isocitrate dehydrogenase (NADP+)] and in the esterases. Ten different loci for NAD-dependent dehydrogenases are electrophoretically monomorphic, as are five different NADH diaphorases from heart muscle and 15 different major proteins from skeletal muscle. It is highly statistically significant that NADP-dependent dehydrogenases and esterases are polymorphic but representatives of several other major classes of enzymes are not. The physiological significance of this polymorphism may be related to the role of these enzymes in growth and detoxication, sheep having been selected by man for faster growth, of wool or of carcass, and for grazing a wide variety of plants.

  20. Oxidant-induced formation of a neutral flavosemiquinone in the Na+-translocating NADH:Quinone oxidoreductase (Na+-NQR) from Vibrio cholerae.

    Science.gov (United States)

    Tao, Minli; Casutt, Marco S; Fritz, Günter; Steuber, Julia

    2008-01-01

    The Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR) from the human pathogen Vibrio cholerae is a respiratory flavo-FeS complex composed of the six subunits NqrA-F. The Na(+)-NQR was produced as His(6)-tagged protein by homologous expression in V. cholerae. The isolated complex contained near-stoichiometric amounts of non-covalently bound FAD (0.78 mol/mol Na(+)-NQR) and riboflavin (0.70 mol/mol Na(+)-NQR), catalyzed NADH-driven Na(+) transport (40 nmol Na(+)min(-1) mg(-1)), and was inhibited by 2-n-heptyl-4-hydroxyquinoline-N-oxide. EPR spectroscopy showed that Na(+)-NQR as isolated contained very low amounts of a neutral flavosemiquinone (10(-3) mol/mol Na(+)-NQR). Reduction with NADH resulted in the formation of an anionic flavosemiquinone (0.10 mol/mol Na(+)-NQR). Subsequent oxidation of the Na(+)-NQR with ubiquinone-1 or O(2) led to the formation of a neutral flavosemiquinone (0.24 mol/mol Na(+)-NQR). We propose that the Na(+)-NQR is fully oxidized in its resting state, and discuss putative schemes of NADH-triggered redox transitions.

  1. Pyruvate dehydrogenase complex and nicotinamide nucleotide transhydrogenase constitute an energy consuming redox circuit

    OpenAIRE

    2015-01-01

    Cellular proteins rely on reversible redox reactions to establish and maintain biological structure and function. How redox catabolic (NAD+:NADH) and anabolic (NADP+:NADPH) processes integrate during metabolism to maintain cellular redox homeostasis however is unknown. The present work identifies a continuously cycling, mitochondrial membrane potential-dependent redox circuit between the pyruvate dehydrogenase complex (PDHC) and nicotinamide nucleotide transhydrogenase (NNT). PDHC is shown to...

  2. Differential pulse voltammetric studies on the effects of Al(Ⅲ) on the lactate dehydrogenase activity

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    In this paper, differential pulse voltammetry (DPV) was applied to study the effects of aluminum Al(Ⅲ) on the lactate dehydrogenase (LDH) activity. Michaelis-Menten constant (KNADHm) and maximum velocity (vmax) in the enzyme promoting catalytic reaction of "pyruvate(Pyr) + NADH + H+ LDH(=) lactate + NAD+" under different conditions by monitoring DPV reduction current of NAD+ were reported.(C) 2007 Shu Ping Bi. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

  3. NAD(+)-linked alcohol dehydrogenase 1 regulates methylglyoxal concentration in Candida albicans.

    Science.gov (United States)

    Kwak, Min-Kyu; Ku, MyungHee; Kang, Sa-Ouk

    2014-04-02

    We purified a fraction that showed NAD(+)-linked methylglyoxal dehydrogenase activity, directly catalyzing methylglyoxal oxidation to pyruvate, which was significantly increased in glutathione-depleted Candida albicans. It also showed NADH-linked methylglyoxal-reducing activity. The fraction was identified as a NAD(+)-linked alcohol dehydrogenase (ADH1) through mass spectrometric analyses. In ADH1-disruptants of both the wild type and glutathione-depleted cells, the intracellular methylglyoxal concentration increased significantly; defects in growth, differentiation, and virulence were observed; and G2-phase arrest was induced.

  4. Stabilized NADH as a Countermeasure for Jet Lag

    Science.gov (United States)

    Kay, Gary G.; Viirre, Erik; Clark, Jonathan

    2001-01-01

    Current remedies for jet lag (phototherapy, melatonin, stimulant, and sedative medications) are limited in efficacy and practicality. The efficacy of a stabilized, sublingual form of reduced nicotin amide adenine dinucleotide (NADH, ENADAlert, Menuco Corp.) as a countermeasure for jet lag was examined. Because NADH increases cellular production of ATP and facilitates dopamine synthesis, it may counteract the effects of jet lag on cognitive functioning and sleepiness. Thirty-five healthy, employed subjects participated in this double-blind, placebo-controlled study. Training and baseline testing were conducted on the West Coast before subjects flew overnight to the East Coast, where they would experience a 3-hour time difference. Upon arrival, individuals were randomly assigned to receive either 20 mg of sublingual stabilized ADH (n=18) or identical placebo tablets (n=17). All participants completed computer-administered tests (including CogScreen7) to assess changes in cognitive functioning, mood, and sleepiness in the morning and afternoon. Jet lag resulted in increased sleepiness for over half the participants and deterioration of cognitive functioning for approximately one third. The morning following the flight, subjects experienced lapses of attention in addition to disruptions in working memory, divided attention, and visual perceptual speed. Individuals who received NADH performed significantly better on 5 of 8 cognitive and psychomotor test measures (P less than or equal to 0.5) and showed a trend for better performance on the other three measures (P less than or equal to .l0). Subjects also reported less sleepiness compared with those who received placebo. No adverse effects were observed with NADH treatment. Stabilized NADH significantly reduced jet lag-induced disruptions of cognitive functioning, was easily administered, and was found to have no adverse side effects.

  5. Furfural reduction mechanism of a zinc-dependent alcohol dehydrogenase from Cupriavidus necator JMP134.

    Science.gov (United States)

    Kang, ChulHee; Hayes, Robert; Sanchez, Emiliano J; Webb, Brian N; Li, Qunrui; Hooper, Travis; Nissen, Mark S; Xun, Luying

    2012-01-01

    FurX is a tetrameric Zn-dependent alcohol dehydrogenase (ADH) from Cupriavidus necator JMP134. The enzyme rapidly reduces furfural with NADH as the reducing power. For the first time among characterized ADHs, the high-resolution structures of all reaction steps were obtained in a time-resolved manner, thereby illustrating the complete catalytic events of NADH-dependent reduction of furfural and the dynamic Zn(2+) coordination among Glu66, water, substrate and product. In the fully closed conformation of the NADH complex, the catalytic turnover proved faster than observed for the partially closed conformation due to an effective proton transfer network. The domain motion triggered by NAD(H) association/dissociation appeared to facilitate dynamic interchanges in Zn(2+) coordination with substrate and product molecules, ultimately increasing the enzymatic turnover rate. NAD(+) dissociation appeared to be a slow process, involving multiple steps in concert with a domain opening and reconfiguration of Glu66. This agrees with the report that the cofactor is not dissociated from FurX during ethanol-dependent reduction of furfural, in which ethanol reduces NAD(+) to NADH that is subsequently used for furfural reduction. © 2011 Blackwell Publishing Ltd.

  6. Cytosolic malate dehydrogenase regulates senescence in human fibroblasts.

    Science.gov (United States)

    Lee, Seung-Min; Dho, So Hee; Ju, Sung-Kyu; Maeng, Jin-Soo; Kim, Jeong-Yoon; Kwon, Ki-Sun

    2012-10-01

    Carbohydrate metabolism changes during cellular senescence. Cytosolic malate dehydrogenase (MDH1) catalyzes the reversible reduction of oxaloacetate to malate at the expense of reduced nicotinamide adenine dinucleotide (NADH). Here, we show that MDH1 plays a critical role in the cellular senescence of human fibroblasts. We observed that the activity of MDH1 was reduced in old human dermal fibroblasts (HDFs) [population doublings (PD) 56], suggesting a link between decreased MDH1 protein levels and aging. Knockdown of MDH1 in young HDFs (PD 20) and the IMR90 human fibroblast cell line resulted in the appearance of significant cellular senescence features, including senescence-associated β-galactosidase staining, flattened and enlarged morphology, increased population doubling time, and elevated p16(INK4A) and p21(CIP1) protein levels. Cytosolic NAD/NADH ratios were decreased in old HDFs to the same extent as in MDH1 knockdown HDFs, suggesting that cytosolic NAD depletion is related to cellular senescence. We found that AMP-activated protein kinase, a sensor of cellular energy, was activated in MDH1 knockdown cells. We also found that sirtuin 1 (SIRT1) deacetylase, a controller of cellular senescence, was decreased in MDH1 knockdown cells. These results indicate that the decrease in MDH1 and subsequent reduction in NAD/NADH ratio, which causes SIRT1 inhibition, is a likely carbohydrate metabolism-controlled cellular senescence mechanism.

  7. Purification of arogenate dehydrogenase from Phenylobacterium immobile.

    Science.gov (United States)

    Mayer, E; Waldner-Sander, S; Keller, B; Keller, E; Lingens, F

    1985-01-07

    Phenylobacterium immobile, a bacterium which is able to degrade the herbicide chloridazon, utilizes for L-tyrosine synthesis arogenate as an obligatory intermediate which is converted in the final biosynthetic step by a dehydrogenase to tyrosine. This enzyme, the arogenate dehydrogenase, has been purified for the first time in a 5-step procedure to homogeneity as confirmed by electrophoresis. The Mr of the enzyme that consists of two identical subunits amounts to 69000 as established by gel electrophoresis after cross-linking the enzyme with dimethylsuberimidate. The Km values were 0.09 mM for arogenate and 0.02 mM for NAD+. The enzyme has a high specificity with respect to its substrate arogenate.

  8. Expression of NAD+-dependent formate dehydrogenase in Enterobacter aerogenes and its involvement in anaerobic metabolism and H2 production.

    Science.gov (United States)

    Lu, Yuan; Zhao, Hongxin; Zhang, Chong; Lai, Qiheng; Wu, Xi; Xing, Xin-Hui

    2009-10-01

    An expression system for NAD(+)-dependent formate dehydrogenase gene (fdh1), from Candida boidinii, was constructed and cloned into Enterobacter aerogenes IAM1183. With the fdh1 expression, the total H(2) yield was attributed to a decrease in activity of the lactate pathway and an increase of the formate pathway flux due to the NADH regeneration. Analysis of the redox state balance and ethanol-to-acetate ratio in the fdhl-expressed strain showed that increased reducing power arose from the reconstruction of NADH regeneration pathway from formate thereby contributing to the improved H(2) production.

  9. Glutamate dehydrogenase from pumpkin cotyledons: characterization and isoenzymes.

    Science.gov (United States)

    Chou, K H; Splittstoesser, W E

    1972-04-01

    Glutamate dehydrogenase from pumpkin (Cucurbita moschata Pior. cultivar Dickinson Field) cotyledons was found in both soluble and particulate fractions with the bulk of the activity in the soluble fraction. Both enzymes used NAD(H) and NADP(H) but NAD(H) was favored. The enzymes were classified as glutamate-NAD oxidoreductase, deaminating (EC 1.4.1.3). Both enzymes were heat stable, had a pH optimum for reductive amination of 8.0, and were inhibited by high concentrations of NH(4) (+) or alpha-ketoglutarate. The soluble enzyme was more sensitive to NH(4) (+) inhibition and was activated by metal ions after ammonium sulfate fractionation while the solubilized particulate enzyme was not. Inhibition by ethylenediaminetetraacetate was restored by several divalent ions and inhibition by p-hydroxymercuribenzoate was reversed by glutathione. Particulate glutamate dehydrogenase showed a greater activity with NADP. The molecular weights of the enzymes are 250,000. Separation of the enzymes by disc gel electrophoresis showed that during germination the soluble isoenzymes increased from 1 to 7 in number, while only one particulate isoenzyme was found at any time. This particulate isoenzyme was identical with one of the soluble isoenzymes. A number of methods indicated that the soluble isoenzymes were not simply removed from the particulate fraction and that true isoenzymes were found.

  10. New biotechnological perspectives of a NADH oxidase variant from Thermus thermophilus HB27 as NAD+-recycling enzyme

    Directory of Open Access Journals (Sweden)

    Rocha-Martín Javier

    2011-11-01

    Full Text Available Abstract Background The number of biotransformations that use nicotinamide recycling systems is exponentially growing. For this reason one of the current challenges in biocatalysis is to develop and optimize more simple and efficient cofactor recycling systems. One promising approach to regenerate NAD+ pools is the use of NADH-oxidases that reduce oxygen to hydrogen peroxide while oxidizing NADH to NAD+. This class of enzymes may be applied to asymmetric reduction of prochiral substrates in order to obtain enantiopure compounds. Results The NADH-oxidase (NOX presented here is a flavoenzyme which needs exogenous FAD or FMN to reach its maximum velocity. Interestingly, this enzyme is 6-fold hyperactivated by incubation at high temperatures (80°C under limiting concentrations of flavin cofactor, a change that remains stable even at low temperatures (37°C. The hyperactivated form presented a high specific activity (37.5 U/mg at low temperatures despite isolation from a thermophile source. Immobilization of NOX onto agarose activated with glyoxyl groups yielded the most stable enzyme preparation (6-fold more stable than the hyperactivated soluble enzyme. The immobilized derivative was able to be reactivated under physiological conditions after inactivation by high solvent concentrations. The inactivation/reactivation cycle could be repeated at least three times, recovering full NOX activity in all cases after the reactivation step. This immobilized catalyst is presented as a recycling partner for a thermophile alcohol dehydrogenase in order to perform the kinetic resolution secondary alcohols. Conclusion We have designed, developed and characterized a heterogeneous and robust biocatalyst which has been used as recycling partner in the kinetic resolution of rac-1-phenylethanol. The high stability along with its capability to be reactivated makes this biocatalyst highly re-useable for cofactor recycling in redox biotransformations.

  11. Electrochemical study in both classical cell and microreactors of flavin adenine dinucleotide as a redox mediator for NADH regeneration

    Energy Technology Data Exchange (ETDEWEB)

    Tzedakis, Theodore, E-mail: tzedakis@chimie.ups-tlse.f [Laboratoire de Genie Chimique, UMR 5503, Universite Paul Sabatier, 31062 Toulouse cedex 04 (France); Cheikhou, Kane [Ecole Superieure Polytechnique de Dakar BP: 16263 Dakar-Fann (Senegal); Jerome, Roche; Karine, Groenen Serrano; Olivier, Reynes [Laboratoire de Genie Chimique, UMR 5503, Universite Paul Sabatier, 31062 Toulouse cedex 04 (France)

    2010-02-28

    The electrochemical reduction of flavin adenine dinucleotide (FAD) is studied in a classical electrochemical cell as well as in two types of microreactors: the first one is a one-channel reactor and the other one, a multichannel filter-press reactor. The ultimate goal is to use the reduced form of flavin (FADH{sub 2}), in the presence of formate dehydrogenase (FDH), in order to continuously regenerate the reduced form of nicotinamide adenine dinucleotide (NADH) for chiral syntheses. Various voltammetric and adsorption measurements were carried out for a better understanding of the redox behavior of the FAD as well as its adsorption on gold. Diffusivity and kinetic electrochemical parameters of FAD were determined.

  12. A water-forming NADH oxidase from Lactobacillus pentosus and its potential application in the regeneration of synthetic biomimetic cofactors

    Directory of Open Access Journals (Sweden)

    Claudia eNowak

    2015-09-01

    Full Text Available The cell-free biocatalytic production of fine chemicals by oxidoreductases has continuously grown over the past years. Since especially dehydrogenases depend on the stoichiometric use of nicotinamide pyridine cofactors, an integrated efficient recycling system is crucial to allow process operation under economic conditions. Lately, the variety of cofactors for biocatalysis was broadened by the utilization of totally synthetic and cheap biomimetics. Though, to date the regeneration has been limited to chemical or electrochemical methods. Here, we report an enzymatic recycling by the flavoprotein NADH-oxidase from Lactobacillus pentosus (LpNox. Since this enzyme has not been described before, we first characterized it in regard to its optimal reaction parameters. We found that the heterologously overexpressed enzyme only contained 13 % FAD. In vitro loading of the enzyme with FAD, resulted in a higher specific activity towards its natural cofactor NADH as well as different nicotinamide derived biomimetics. Apart from the enzymatic recycling, which gives water as a by-product by transferring four electrons onto oxygen, unbound FAD can also catalyse the oxidation of biomimetic cofactors. Here a two electron process takes place yielding H2O2 instead. The enzymatic and chemical recycling was compared in regard to reaction kinetics for the natural and biomimetic cofactors. With LpNox and FAD, two recycling strategies for biomimetic cofactors are described with either water or hydrogen peroxide as a by-product.

  13. A water-forming NADH oxidase from Lactobacillus pentosus suitable for the regeneration of synthetic biomimetic cofactors.

    Science.gov (United States)

    Nowak, Claudia; Beer, Barbara; Pick, André; Roth, Teresa; Lommes, Petra; Sieber, Volker

    2015-01-01

    The cell-free biocatalytic production of fine chemicals by oxidoreductases has continuously grown over the past years. Since especially dehydrogenases depend on the stoichiometric use of nicotinamide pyridine cofactors, an integrated efficient recycling system is crucial to allow process operation under economic conditions. Lately, the variety of cofactors for biocatalysis was broadened by the utilization of totally synthetic and cheap biomimetics. Though, to date the regeneration has been limited to chemical or electrochemical methods. Here, we report an enzymatic recycling by the flavoprotein NADH-oxidase from Lactobacillus pentosus (LpNox). Since this enzyme has not been described before, we first characterized it in regard to its optimal reaction parameters. We found that the heterologously overexpressed enzyme only contained 13% FAD. In vitro loading of the enzyme with FAD, resulted in a higher specific activity towards its natural cofactor NADH as well as different nicotinamide derived biomimetics. Apart from the enzymatic recycling, which gives water as a by-product by transferring four electrons onto oxygen, unbound FAD can also catalyze the oxidation of biomimetic cofactors. Here a two electron process takes place yielding H2O2 instead. The enzymatic and chemical recycling was compared in regard to reaction kinetics for the natural and biomimetic cofactors. With LpNox and FAD, two recycling strategies for biomimetic cofactors are described with either water or hydrogen peroxide as by-product.

  14. AcEST: DK949667 [AcEST

    Lifescience Database Archive (English)

    Full Text Available WC1_9ASTR NADH dehydrogenase subunit I OS=Onoseris ... 250 4e-65 tr|B0Z5I1|B0Z5I1_OENPA NADH dehydrogenase s...PVIEDYTIRTILNS 161 >tr|B2XWC1|B2XWC1_9ASTR NADH dehydrogenase subunit I OS=Onoseris hastata GN=ndhI PE=4 SV=

  15. Investigations concerning the determination of NADH concentrations using optical biopsy

    Science.gov (United States)

    Beuthan, Juergen; Bocher, Thomas; Minet, Olaf; Roggan, Andre; Schmitt, Isabella; Weber, A.; Mueller, Gerhard J.

    1994-05-01

    The intrinsic NADH autofluorescence intensity of biological tissue depends on the local, cellular concentration of this coenzyme. It plays a dominant role in the Krebs-Cycle and therefore serves as indicator for the vitality of the observed cells. Due to individually and locally varying boundary conditions and optical tissue properties, which are scattering coefficients, absorption coefficients and g-factors the fluorescence signal needs to be rescaled. One possible rescaling method is the theoretical derived Photon Migration Theory. Our new rescaling method is partly based on measurements and partly theoretical derived. By using the 4 information channels: LIF time-resolved signal, biochemical concentration measurements, Monte Carlo simulations with optical parameters and microscopic investigations we demonstrate that simultaneous detection of the fluorescence and the backscattering signal can easily and accurately provide rescaled, quantitative values for the NADH concentrations.

  16. Involvement of NADH Oxidase in Biofilm Formation in Streptococcus sanguinis.

    Directory of Open Access Journals (Sweden)

    Xiuchun Ge

    Full Text Available Biofilms play important roles in microbial communities and are related to infectious diseases. Here, we report direct evidence that a bacterial nox gene encoding NADH oxidase is involved in biofilm formation. A dramatic reduction in biofilm formation was observed in a Streptococcus sanguinis nox mutant under anaerobic conditions without any decrease in growth. The membrane fluidity of the mutant bacterial cells was found to be decreased and the fatty acid composition altered, with increased palmitic acid and decreased stearic acid and vaccenic acid. Extracellular DNA of the mutant was reduced in abundance and bacterial competence was suppressed. Gene expression analysis in the mutant identified two genes with altered expression, gtfP and Idh, which were found to be related to biofilm formation through examination of their deletion mutants. NADH oxidase-related metabolic pathways were analyzed, further clarifying the function of this enzyme in biofilm formation.

  17. Production of superoxide/hydrogen peroxide by the mitochondrial 2-oxoadipate dehydrogenase complex.

    Science.gov (United States)

    Goncalves, Renata L S; Bunik, Victoria I; Brand, Martin D

    2016-02-01

    In humans, mutations in dehydrogenase E1 and transketolase domain containing 1 (DHTKD1) are associated with neurological abnormalities and accumulation of 2-oxoadipate, 2-aminoadipate, and reactive oxygen species. The protein encoded by DHTKD1 has sequence and structural similarities to 2-oxoglutarate dehydrogenase, and the 2-oxoglutarate dehydrogenase complex can produce superoxide/H2O2 at high rates. The DHTKD1 enzyme is hypothesized to catalyze the oxidative decarboxylation of 2-oxoadipate, a shared intermediate of the degradative pathways for tryptophan, lysine and hydroxylysine. Here, we show that rat skeletal muscle mitochondria can produce superoxide/H2O2 at high rates when given 2-oxoadipate. We identify the putative mitochondrial 2-oxoadipate dehydrogenase complex as one of the sources and characterize the conditions that favor its superoxide/H2O2 production. Rates increased at higher NAD(P)H/NAD(P)(+) ratios and were higher at each NAD(P)H/NAD(P)(+) ratio when 2-oxoadipate was present, showing that superoxide/H2O2 was produced during the forward reaction from 2-oxoadipate, but not in the reverse reaction from NADH in the absence of 2-oxoadipate. The maximum capacity of the 2-oxoadipate dehydrogenase complex for production of superoxide/H2O2 is comparable to that of site IF of complex I, and seven, four and almost two-fold lower than the capacities of the 2-oxoglutarate, pyruvate and branched-chain 2-oxoacid dehydrogenase complexes, respectively. Regulation by ADP and ATP of H2O2 production driven by 2-oxoadipate was very different from that driven by 2-oxoglutarate, suggesting that site AF of the 2-oxoadipate dehydrogenase complex is a new source of superoxide/H2O2 associated with the NADH isopotential pool in mitochondria.

  18. NADH fluorescence imaging of isolated biventricular working rabbit hearts.

    Science.gov (United States)

    Asfour, Huda; Wengrowski, Anastasia M; Jaimes, Rafael; Swift, Luther M; Kay, Matthew W

    2012-07-24

    Since its inception by Langendorff(1), the isolated perfused heart remains a prominent tool for studying cardiac physiology(2). However, it is not well-suited for studies of cardiac metabolism, which require the heart to perform work within the context of physiologic preload and afterload pressures. Neely introduced modifications to the Langendorff technique to establish appropriate left ventricular (LV) preload and afterload pressures(3). The model is known as the isolated LV working heart model and has been used extensively to study LV performance and metabolism(4-6). This model, however, does not provide a properly loaded right ventricle (RV). Demmy et al. first reported a biventricular model as a modification of the LV working heart model(7, 8). They found that stroke volume, cardiac output, and pressure development improved in hearts converted from working LV mode to biventricular working mode(8). A properly loaded RV also diminishes abnormal pressure gradients across the septum to improve septal function. Biventricular working hearts have been shown to maintain aortic output, pulmonary flow, mean aortic pressure, heart rate, and myocardial ATP levels for up to 3 hours(8). When studying the metabolic effects of myocardial injury, such as ischemia, it is often necessary to identify the location of the affected tissue. This can be done by imaging the fluorescence of NADH (the reduced form of nicotinamide adenine dinucleotide)(9-11), a coenzyme found in large quantities in the mitochondria. NADH fluorescence (fNADH) displays a near linearly inverse relationship with local oxygen concentration(12) and provides a measure of mitochondrial redox state(13). fNADH imaging during hypoxic and ischemic conditions has been used as a dye-free method to identify hypoxic regions(14, 15) and to monitor the progression of hypoxic conditions over time(10). The objective of the method is to monitor the mitochondrial redox state of biventricular working hearts during protocols

  19. Engineering of alanine dehydrogenase from Bacillus subtilis for novel cofactor specificity.

    Science.gov (United States)

    Lerchner, Alexandra; Jarasch, Alexander; Skerra, Arne

    2016-09-01

    The l-alanine dehydrogenase of Bacillus subtilis (BasAlaDH), which is strictly dependent on NADH as redox cofactor, efficiently catalyzes the reductive amination of pyruvate to l-alanine using ammonia as amino group donor. To enable application of BasAlaDH as regenerating enzyme in coupled reactions with NADPH-dependent alcohol dehydrogenases, we alterated its cofactor specificity from NADH to NADPH via protein engineering. By introducing two amino acid exchanges, D196A and L197R, high catalytic efficiency for NADPH was achieved, with kcat /KM  = 54.1 µM(-1)  Min(-1) (KM  = 32 ± 3 µM; kcat  = 1,730 ± 39 Min(-1) ), almost the same as the wild-type enzyme for NADH (kcat /KM  = 59.9 µM(-1)  Min(-1) ; KM  = 14 ± 2 µM; kcat  = 838 ± 21 Min(-1) ). Conversely, recognition of NADH was much diminished in the mutated enzyme (kcat /KM  = 3 µM(-1)  Min(-1) ). BasAlaDH(D196A/L197R) was applied in a coupled oxidation/transamination reaction of the chiral dicyclic dialcohol isosorbide to its diamines, catalyzed by Ralstonia sp. alcohol dehydrogenase and Paracoccus denitrificans ω-aminotransferase, thus allowing recycling of the two cosubstrates NADP(+) and l-Ala. An excellent cofactor regeneration with recycling factors of 33 for NADP(+) and 13 for l-Ala was observed with the engineered BasAlaDH in a small-scale biocatalysis experiment. This opens a biocatalytic route to novel building blocks for industrial high-performance polymers.

  20. Water-insoluble material from apple pomace makes changes in intracellular NAD⁺/NADH ratio and pyrophosphate content and stimulates fermentative production of hydrogen.

    Science.gov (United States)

    Sato, Osamu; Suzuki, Yuma; Sato, Yuki; Sasaki, Shinsuke; Sonoki, Tomonori

    2015-05-01

    Apple pomace is one of the major agricultural residues in Aomori prefecture, Japan, and it would be useful to develop effective applications for it. As apple pomace contains easily fermentable sugars such as glucose, fructose and sucrose, it can be used as a feedstock for the fermentation of fuels and chemicals. We previously isolated a new hydrogen-producing bacterium, Clostridium beijerinckii HU-1, which could produce H2 at a production rate of 14.5 mmol of H2/L/h in a fed-batch culture at 37 °C, pH 6.0. In this work we found that the HU-1 strain produces H2 at an approximately 20% greater rate when the fermentation medium contains the water-insoluble material from apple pomace. The water-insoluble material from apple pomace caused a metabolic shift that stimulated H2 production. HU-1 showed a decrease of lactate production, which consumes NADH, accompanied by an increase of the intracellular pyrophosphate content, which is an inhibitor of lactate dehydrogenase. The intracellular NAD(+)/NADH ratios of HU-1 during H2 fermentation were maintained in a more reductive state than those observed without the addition of the water insoluble material. To correct the abnormal intracellular redox balance, caused by the repression of lactate production, H2 production with NADH oxidation must be stimulated.

  1. Molecular cloning, characterization and regulation of two different NADH-glutamate synthase cDNAs in bean nodules

    Science.gov (United States)

    NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) is a key enzyme in primary ammonia assimilation in bean (Phaseolus vulgaris L.) nodules. Two different types of cDNA clones of PvNADH-GOGAT were isolated from two independent nodule cDNA libraries. The full-length cDNA clones of PvNADH-GOGA...

  2. AcEST: BP914956 [AcEST

    Lifescience Database Archive (English)

    Full Text Available HEMI NADH dehydrogenase subunit 4 OS=Geocoris ... 33 7.0 tr|B8MHC1|B8MHC1_9EURO Cell cycle control protein (...SMC2|B7SMC2_9HEMI NADH dehydrogenase subunit 4 OS=Geocoris pallidipennis GN=ND4 PE=4 SV=1 Length = 438 Score

  3. NCBI nr-aa BLAST: CBRC-OPRI-01-1140 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OPRI-01-1140 ref|YP_002519420.1| NADH dehydrogenase subunit 2 [Bombus hypocrita sapporo...ensis] gb|ABY75171.1| NADH dehydrogenase subunit 2 [Bombus hypocrita sapporoensis] YP_002519420.1 0.30 22% ...

  4. NCBI nr-aa BLAST: CBRC-PHAM-01-1493 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PHAM-01-1493 ref|YP_002519420.1| NADH dehydrogenase subunit 2 [Bombus hypocrita sapporo...ensis] gb|ABY75171.1| NADH dehydrogenase subunit 2 [Bombus hypocrita sapporoensis] YP_002519420.1 0.13 22% ...

  5. NCBI nr-aa BLAST: CBRC-LAFR-01-0694 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-0694 ref|NP_066533.1| NADH dehydrogenase subunit 2 [Naegleria gruberi]... gb|AAG17811.1|AF288092_36 NADH dehydrogenase subunit 2 [Naegleria gruberi] NP_066533.1 0.66 25% ...

  6. NCBI nr-aa BLAST: CBRC-OPRI-01-0587 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OPRI-01-0587 ref|YP_001936231.1| NADH dehydrogenase subunit 5 [Phalangium opil...io] gb|ACA66085.1| NADH dehydrogenase subunit 5 [Phalangium opilio] YP_001936231.1 0.30 23% ...

  7. NCBI nr-aa BLAST: CBRC-DNOV-01-1200 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-1200 ref|YP_784045.1| NADH dehydrogenase subunit 1 [Scutigerella cause...yae] gb|ABF93312.1| NADH dehydrogenase subunit 1 [Scutigerella causeyae] YP_784045.1 1.6 23% ...

  8. NCBI nr-aa BLAST: CBRC-TBEL-01-0894 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TBEL-01-0894 ref|YP_784041.1| NADH dehydrogenase subunit 4 [Scutigerella cause...yae] gb|ABF93309.1| NADH dehydrogenase subunit 4 [Scutigerella causeyae] YP_784041.1 0.050 28% ...

  9. NCBI nr-aa BLAST: CBRC-MEUG-01-1375 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MEUG-01-1375 ref|YP_002261391.1| NADH dehydrogenase subunit 2 [Troglophilus neglect...us] gb|ACG59335.1| NADH dehydrogenase subunit 2 [Troglophilus neglectus] YP_002261391.1 0.31 30% ...

  10. NCBI nr-aa BLAST: CBRC-OPRI-01-0222 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OPRI-01-0222 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.94 23% ...

  11. NCBI nr-aa BLAST: CBRC-TTRU-01-0312 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0312 ref|YP_002317278.1| NADH dehydrogenase subunit 5 [Steganacarus magnu...s] gb|ACH41151.1| NADH dehydrogenase subunit 5 [Steganacarus magnus] YP_002317278.1 0.002 27% ...

  12. NCBI nr-aa BLAST: CBRC-OPRI-01-0769 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OPRI-01-0769 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.11 23% ...

  13. NCBI nr-aa BLAST: CBRC-MDOM-03-0070 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-03-0070 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.016 31% ...

  14. NCBI nr-aa BLAST: CBRC-TTRU-01-0165 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0165 ref|YP_002317278.1| NADH dehydrogenase subunit 5 [Steganacarus magnu...s] gb|ACH41151.1| NADH dehydrogenase subunit 5 [Steganacarus magnus] YP_002317278.1 3e-04 30% ...

  15. NCBI nr-aa BLAST: CBRC-MDOM-04-0076 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-04-0076 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.36 23% ...

  16. NCBI nr-aa BLAST: CBRC-MDOM-01-0134 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0134 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.029 25% ...

  17. NCBI nr-aa BLAST: CBRC-MDOM-01-0273 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0273 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.12 25% ...

  18. NCBI nr-aa BLAST: CBRC-MDOM-05-0011 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-05-0011 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.015 24% ...

  19. NCBI nr-aa BLAST: CBRC-MDOM-05-0015 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-05-0015 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.81 24% ...

  20. NCBI nr-aa BLAST: CBRC-MDOM-08-0245 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-08-0245 ref|YP_002317278.1| NADH dehydrogenase subunit 5 [Steganacarus magnu...s] gb|ACH41151.1| NADH dehydrogenase subunit 5 [Steganacarus magnus] YP_002317278.1 0.16 30% ...

  1. NCBI nr-aa BLAST: CBRC-OPRI-01-0087 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OPRI-01-0087 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.19 21% ...

  2. NCBI nr-aa BLAST: CBRC-MDOM-03-0035 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-03-0035 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.17 24% ...

  3. NCBI nr-aa BLAST: CBRC-MDOM-07-0060 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-07-0060 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.046 24% ...

  4. NCBI nr-aa BLAST: CBRC-MDOM-05-0049 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-05-0049 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.12 25% ...

  5. NCBI nr-aa BLAST: CBRC-MDOM-08-0115 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-08-0115 ref|YP_002317279.1| NADH dehydrogenase subunit 4 [Steganacarus magnu...s] gb|ACH41152.1| NADH dehydrogenase subunit 4 [Steganacarus magnus] YP_002317279.1 0.005 26% ...

  6. NCBI nr-aa BLAST: CBRC-TTRU-01-0146 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0146 ref|YP_001837118.1| NADH dehydrogenase subunit 4 [Walchia hayashi...i] dbj|BAG24174.1| NADH dehydrogenase subunit 4 [Walchia hayashii] YP_001837118.1 0.019 23% ...

  7. NCBI nr-aa BLAST: CBRC-STRI-01-2357 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-2357 ref|YP_003097101.1| NADH dehydrogenase subunit 6 [Angiostrongylus canton...ensis] gb|ACT88784.1| NADH dehydrogenase subunit 6 [Angiostrongylus cantonensis] YP_003097101.1 0.16 28% ...

  8. NCBI nr-aa BLAST: CBRC-TTRU-01-0610 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0610 ref|YP_003097108.1| NADH dehydrogenase subunit 4 [Angiostrongylus canton...ensis] gb|ACT88791.1| NADH dehydrogenase subunit 4 [Angiostrongylus cantonensis] YP_003097108.1 0.021 25% ...

  9. NCBI nr-aa BLAST: CBRC-TTRU-01-0527 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0527 ref|YP_003097108.1| NADH dehydrogenase subunit 4 [Angiostrongylus canton...ensis] gb|ACT88791.1| NADH dehydrogenase subunit 4 [Angiostrongylus cantonensis] YP_003097108.1 0.88 27% ...

  10. NCBI nr-aa BLAST: CBRC-DNOV-01-0578 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0578 ref|YP_001256912.1| NADH dehydrogenase subunit 1 [Reticulitermes santon...ensis] gb|ABN10443.1| NADH dehydrogenase subunit 1 [Reticulitermes santonensis] YP_001256912.1 2.4 23% ...

  11. NCBI nr-aa BLAST: CBRC-DNOV-01-2359 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2359 ref|YP_001256912.1| NADH dehydrogenase subunit 1 [Reticulitermes santon...ensis] gb|ABN10443.1| NADH dehydrogenase subunit 1 [Reticulitermes santonensis] YP_001256912.1 2.1 23% ...

  12. NCBI nr-aa BLAST: CBRC-CELE-01-0017 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CELE-01-0017 ref|YP_537103.1| NADH dehydrogenase subunit 2 [Paracoccidioides brasil...iensis] gb|AAY30327.1| NADH dehydrogenase subunit 2 [Paracoccidioides brasiliensis] YP_537103.1 0.022 21% ...

  13. NCBI nr-aa BLAST: CBRC-CBRE-01-1153 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CBRE-01-1153 ref|YP_073309.1| NADH dehydrogenase subunit 2 [Schizaphis graminu...m] gb|AAS00827.1| NADH dehydrogenase subunit 2 [Schizaphis graminum] YP_073309.1 0.034 21% ...

  14. NCBI nr-aa BLAST: CBRC-TTRU-01-1218 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1218 ref|YP_073303.1| NADH dehydrogenase subunit 5 [Schizaphis graminu...m] gb|AAS00821.1| NADH dehydrogenase subunit 5 [Schizaphis graminum] YP_073303.1 0.007 24% ...

  15. NCBI nr-aa BLAST: CBRC-TTRU-01-0389 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0389 ref|YP_073303.1| NADH dehydrogenase subunit 5 [Schizaphis graminu...m] gb|AAS00821.1| NADH dehydrogenase subunit 5 [Schizaphis graminum] YP_073303.1 0.003 26% ...

  16. NCBI nr-aa BLAST: CBRC-CBRE-01-1139 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CBRE-01-1139 ref|YP_073309.1| NADH dehydrogenase subunit 2 [Schizaphis graminu...m] gb|AAS00827.1| NADH dehydrogenase subunit 2 [Schizaphis graminum] YP_073309.1 0.007 22% ...

  17. NCBI nr-aa BLAST: CBRC-TTRU-01-1105 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1105 ref|YP_073303.1| NADH dehydrogenase subunit 5 [Schizaphis graminu...m] gb|AAS00821.1| NADH dehydrogenase subunit 5 [Schizaphis graminum] YP_073303.1 9e-04 25% ...

  18. NCBI nr-aa BLAST: CBRC-TTRU-01-1284 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1284 ref|YP_073303.1| NADH dehydrogenase subunit 5 [Schizaphis graminu...m] gb|AAS00821.1| NADH dehydrogenase subunit 5 [Schizaphis graminum] YP_073303.1 0.005 24% ...

  19. NCBI nr-aa BLAST: CBRC-DNOV-01-0511 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0511 ref|YP_087066.1|ND5_17929 NADH dehydrogenase subunit 5 [Megabalan...us volcano] dbj|BAD44768.1| NADH dehydrogenase subunit 5 [Megabalanus volcano] YP_087066.1 0.56 23% ...

  20. NCBI nr-aa BLAST: CBRC-OSAT-07-0033 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OSAT-07-0033 ref|YP_087065.1|ND4_17929 NADH dehydrogenase subunit 4 [Megabalan...us volcano] dbj|BAD44767.1| NADH dehydrogenase subunit 4 [Megabalanus volcano] YP_087065.1 2.4 30% ...

  1. NCBI nr-aa BLAST: CBRC-BTAU-01-2342 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-BTAU-01-2342 ref|YP_087065.1|ND4_17929 NADH dehydrogenase subunit 4 [Megabalan...us volcano] dbj|BAD44767.1| NADH dehydrogenase subunit 4 [Megabalanus volcano] YP_087065.1 0.003 21% ...

  2. NCBI nr-aa BLAST: CBRC-RNOR-03-0485 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-03-0485 gb|AAY43510.1| NADH dehydrogenase subunit 2 [Milvus milvus fascii...cauda] gb|AAY43514.1| NADH dehydrogenase subunit 2 [Milvus milvus fasciicauda] AAY43510.1 0.19 25% ...

  3. NCBI nr-aa BLAST: CBRC-TTRU-01-0411 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0411 ref|YP_003097082.1| NADH dehydrogenase subunit 5 [Rhopalomyia pom...um] gb|ACT80211.1| NADH dehydrogenase subunit 5 [Rhopalomyia pomum] YP_003097082.1 0.002 23% ...

  4. NCBI nr-aa BLAST: CBRC-MDOM-01-0447 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0447 ref|YP_003097087.1| NADH dehydrogenase subunit 1 [Rhopalomyia pom...um] gb|ACT80216.1| NADH dehydrogenase subunit 1 [Rhopalomyia pomum] YP_003097087.1 0.060 30% ...

  5. NCBI nr-aa BLAST: CBRC-PCAP-01-0672 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PCAP-01-0672 ref|YP_003097087.1| NADH dehydrogenase subunit 1 [Rhopalomyia pom...um] gb|ACT80216.1| NADH dehydrogenase subunit 1 [Rhopalomyia pomum] YP_003097087.1 0.098 26% ...

  6. NCBI nr-aa BLAST: CBRC-TTRU-01-1013 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1013 ref|YP_003097082.1| NADH dehydrogenase subunit 5 [Rhopalomyia pom...um] gb|ACT80211.1| NADH dehydrogenase subunit 5 [Rhopalomyia pomum] YP_003097082.1 0.006 23% ...

  7. NCBI nr-aa BLAST: CBRC-TTRU-01-0140 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0140 ref|YP_003097082.1| NADH dehydrogenase subunit 5 [Rhopalomyia pom...um] gb|ACT80211.1| NADH dehydrogenase subunit 5 [Rhopalomyia pomum] YP_003097082.1 0.026 23% ...

  8. NCBI nr-aa BLAST: CBRC-TTRU-01-1203 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1203 ref|YP_003097082.1| NADH dehydrogenase subunit 5 [Rhopalomyia pom...um] gb|ACT80211.1| NADH dehydrogenase subunit 5 [Rhopalomyia pomum] YP_003097082.1 0.002 23% ...

  9. NCBI nr-aa BLAST: CBRC-TTRU-01-1187 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1187 ref|YP_002970996.1| NADH dehydrogenase subunit 6 [Loligo opalesce...ns] gb|ACS12932.1| NADH dehydrogenase subunit 6 [Loligo opalescens] YP_002970996.1 0.12 23% ...

  10. NCBI nr-aa BLAST: CBRC-CBRE-01-1180 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CBRE-01-1180 ref|NP_051147.1| NADH dehydrogenase subunit 2 [Cafeteria roenberg...ensis] gb|AAF05798.1|AF193903_21 NADH dehydrogenase subunit 2 [Cafeteria roenbergensis] NP_051147.1 2e-05 24% ...

  11. NCBI nr-aa BLAST: CBRC-DNOV-01-2064 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2064 ref|NP_051147.1| NADH dehydrogenase subunit 2 [Cafeteria roenberg...ensis] gb|AAF05798.1|AF193903_21 NADH dehydrogenase subunit 2 [Cafeteria roenbergensis] NP_051147.1 0.009 24% ...

  12. NCBI nr-aa BLAST: CBRC-CREM-01-0003 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CREM-01-0003 ref|NP_051147.1| NADH dehydrogenase subunit 2 [Cafeteria roenberg...ensis] gb|AAF05798.1|AF193903_21 NADH dehydrogenase subunit 2 [Cafeteria roenbergensis] NP_051147.1 1e-04 27% ...

  13. NCBI nr-aa BLAST: CBRC-XTRO-01-2845 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-2845 ref|NP_066350.1| NADH dehydrogenase subunit 2 [Malawimonas jakobi...formis] gb|AAG13717.1|AF295546_43 NADH dehydrogenase subunit 2 [Malawimonas jakobiformis] NP_066350.1 0.71 22% ...

  14. NCBI nr-aa BLAST: CBRC-TTRU-01-0923 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0923 ref|YP_448919.1| NADH dehydrogenase subunit 1 [Sclerophasma paresisense...] gb|ABB81903.1| NADH dehydrogenase subunit 1 [Sclerophasma paresisense] YP_448919.1 0.003 26% ...

  15. NCBI nr-aa BLAST: CBRC-TTRU-01-0023 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0023 ref|YP_448919.1| NADH dehydrogenase subunit 1 [Sclerophasma paresisense...] gb|ABB81903.1| NADH dehydrogenase subunit 1 [Sclerophasma paresisense] YP_448919.1 0.040 27% ...

  16. NCBI nr-aa BLAST: CBRC-TSYR-01-0210 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TSYR-01-0210 ref|YP_448915.1| NADH dehydrogenase subunit 4 [Sclerophasma paresisense...] gb|ABB81899.1| NADH dehydrogenase subunit 4 [Sclerophasma paresisense] YP_448915.1 0.004 23% ...

  17. NCBI nr-aa BLAST: CBRC-TSYR-01-0107 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TSYR-01-0107 ref|YP_448915.1| NADH dehydrogenase subunit 4 [Sclerophasma paresisense...] gb|ABB81899.1| NADH dehydrogenase subunit 4 [Sclerophasma paresisense] YP_448915.1 0.035 22% ...

  18. NCBI nr-aa BLAST: CBRC-LAFR-01-0838 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-0838 ref|YP_227573.1| NADH dehydrogenase subunit 4 [Candida orthopsilos...is] gb|AAX73018.1| NADH dehydrogenase subunit 4 [Candida orthopsilosis] YP_227573.1 1e-105 80% ...

  19. NCBI nr-aa BLAST: CBRC-LAFR-01-0838 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-0838 ref|NP_943641.2| NADH dehydrogenase subunit 4 [Candida parapsilos...is] emb|CAE54604.2| NADH dehydrogenase subunit 4 [Candida parapsilosis] NP_943641.2 1e-106 80% ...

  20. NCBI nr-aa BLAST: CBRC-LAFR-01-0838 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-0838 ref|YP_227558.1| NADH dehydrogenase subunit 4 [Candida metapsilos...is] gb|AAX73033.1| NADH dehydrogenase subunit 4 [Candida metapsilosis] YP_227558.1 1e-105 80% ...

  1. NCBI nr-aa BLAST: CBRC-DNOV-01-2024 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2024 ref|YP_514788.1| NADH dehydrogenase subunit 5 [Sepioteuthis lesson...iana] dbj|BAE80054.1| NADH dehydrogenase subunit 5 [Sepioteuthis lessoniana] YP_514788.1 0.26 28% ...

  2. NCBI nr-aa BLAST: CBRC-TTRU-01-0536 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0536 ref|NP_063966.2| NADH dehydrogenase subunit 2 [Paragonimus westerman...i] gb|AAM98248.1| NADH dehydrogenase subunit 2 [Paragonimus westermani] NP_063966.2 0.026 26% ...

  3. AcEST: DK956396 [AcEST

    Lifescience Database Archive (English)

    Full Text Available ---GGSFFLSVSCLFSFLFLGGLFWCYSYSWCHCWCHMMLSSSAVLV 177 >tr|Q8M676|Q8M676_9TREM NADH dehydrogenase subunit 2 OS=Paragonimus westerman...FF----LSSGGAFFLNVVCVLSFLFLGGLFWCYSYSWCHCWCHMMLSSSA 174 >tr|Q8HN33|Q8HN33_9TREM NADH dehydrogenase subunit 2 OS=Paragonimus westerman

  4. NCBI nr-aa BLAST: CBRC-CBRI-01-0004 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CBRI-01-0004 ref|NP_063966.2| NADH dehydrogenase subunit 2 [Paragonimus westerman...i] gb|AAM98248.1| NADH dehydrogenase subunit 2 [Paragonimus westermani] NP_063966.2 0.068 28% ...

  5. NCBI nr-aa BLAST: CBRC-TTRU-01-1031 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1031 ref|NP_063966.2| NADH dehydrogenase subunit 2 [Paragonimus westerman...i] gb|AAM98248.1| NADH dehydrogenase subunit 2 [Paragonimus westermani] NP_063966.2 2.6 33% ...

  6. NCBI nr-aa BLAST: CBRC-TTRU-01-1259 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1259 ref|NP_059467.1| NADH dehydrogenase subunit 6 [Paragonimus westerman...i] gb|AAF73395.1|AF219379_9 NADH dehydrogenase subunit 6 [Paragonimus westermani] NP_059467.1 0.076 27% ...

  7. NCBI nr-aa BLAST: CBRC-DNOV-01-0366 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0366 ref|NP_059467.1| NADH dehydrogenase subunit 6 [Paragonimus westerman...i] gb|AAF73395.1|AF219379_9 NADH dehydrogenase subunit 6 [Paragonimus westermani] NP_059467.1 0.75 24% ...

  8. NCBI nr-aa BLAST: CBRC-RNOR-21-0156 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-21-0156 ref|NP_063966.2| NADH dehydrogenase subunit 2 [Paragonimus westerman...i] gb|AAM98248.1| NADH dehydrogenase subunit 2 [Paragonimus westermani] NP_063966.2 0.70 25% ...

  9. NCBI nr-aa BLAST: CBRC-TTRU-01-0824 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0824 ref|YP_087051.1| NADH dehydrogenase subunit 6 [Tetraleurodes acacia...e] gb|AAU14156.1| NADH dehydrogenase subunit 6 [Tetraleurodes acaciae] YP_087051.1 0.14 23% ...

  10. NCBI nr-aa BLAST: CBRC-TTRU-01-0873 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0873 ref|YP_087051.1| NADH dehydrogenase subunit 6 [Tetraleurodes acacia...e] gb|AAU14156.1| NADH dehydrogenase subunit 6 [Tetraleurodes acaciae] YP_087051.1 0.018 30% ...

  11. NCBI nr-aa BLAST: CBRC-TTRU-01-1194 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1194 ref|YP_087051.1| NADH dehydrogenase subunit 6 [Tetraleurodes acacia...e] gb|AAU14156.1| NADH dehydrogenase subunit 6 [Tetraleurodes acaciae] YP_087051.1 0.009 30% ...

  12. NCBI nr-aa BLAST: CBRC-DNOV-01-2430 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2430 ref|YP_087056.1| NADH dehydrogenase subunit 2 [Tetraleurodes acacia...e] gb|AAU14160.1| NADH dehydrogenase subunit 2 [Tetraleurodes acaciae] YP_087056.1 1.2 24% ...

  13. NCBI nr-aa BLAST: CBRC-TTRU-01-1250 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1250 ref|YP_087051.1| NADH dehydrogenase subunit 6 [Tetraleurodes acacia...e] gb|AAU14156.1| NADH dehydrogenase subunit 6 [Tetraleurodes acaciae] YP_087051.1 0.020 29% ...

  14. NCBI nr-aa BLAST: CBRC-TTRU-01-1229 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1229 ref|YP_087051.1| NADH dehydrogenase subunit 6 [Tetraleurodes acacia...e] gb|AAU14156.1| NADH dehydrogenase subunit 6 [Tetraleurodes acaciae] YP_087051.1 0.011 28% ...

  15. NCBI nr-aa BLAST: CBRC-DNOV-01-1740 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-1740 ref|YP_087056.1| NADH dehydrogenase subunit 2 [Tetraleurodes acacia...e] gb|AAU14160.1| NADH dehydrogenase subunit 2 [Tetraleurodes acaciae] YP_087056.1 0.009 24% ...

  16. NCBI nr-aa BLAST: CBRC-TTRU-01-1377 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1377 ref|YP_087051.1| NADH dehydrogenase subunit 6 [Tetraleurodes acacia...e] gb|AAU14156.1| NADH dehydrogenase subunit 6 [Tetraleurodes acaciae] YP_087051.1 0.004 31% ...

  17. NCBI nr-aa BLAST: CBRC-DNOV-01-0383 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0383 ref|YP_087056.1| NADH dehydrogenase subunit 2 [Tetraleurodes acacia...e] gb|AAU14160.1| NADH dehydrogenase subunit 2 [Tetraleurodes acaciae] YP_087056.1 0.22 23% ...

  18. NCBI nr-aa BLAST: CBRC-TTRU-01-0598 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0598 ref|YP_087051.1| NADH dehydrogenase subunit 6 [Tetraleurodes acacia...e] gb|AAU14156.1| NADH dehydrogenase subunit 6 [Tetraleurodes acaciae] YP_087051.1 0.015 30% ...

  19. NCBI nr-aa BLAST: CBRC-TTRU-01-1076 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1076 ref|YP_087051.1| NADH dehydrogenase subunit 6 [Tetraleurodes acacia...e] gb|AAU14156.1| NADH dehydrogenase subunit 6 [Tetraleurodes acaciae] YP_087051.1 0.002 31% ...

  20. NCBI nr-aa BLAST: CBRC-SARA-01-0430 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-SARA-01-0430 ref|NP_878187.1| NADH dehydrogenase subunit 1 [Fejervarya limnoch...aris] gb|AAO12107.1| NADH dehydrogenase subunit 1 [Fejervarya limnocharis] NP_878187.1 2.5 29% ...

  1. NCBI nr-aa BLAST: CBRC-OCUN-01-0083 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0083 ref|NP_044800.1| NADH dehydrogenase subunit 5 [Reclinomonas america...na] gb|AAD11915.1| NADH dehydrogenase subunit 5 [Reclinomonas americana] NP_044800.1 3e-82 47% ...

  2. NCBI nr-aa BLAST: CBRC-CFAM-05-0058 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CFAM-05-0058 ref|YP_025917.1| NADH dehydrogenase subunit 5 [Xiphinema american...um] gb|AAQ75781.1| NADH dehydrogenase subunit 5 [Xiphinema americanum] YP_025917.1 5.0 20% ...

  3. NCBI nr-aa BLAST: CBRC-CBRI-07-0000 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CBRI-07-0000 ref|NP_579967.2| NADH dehydrogenase subunit 5 [Necator americanus...] emb|CAD10452.2| NADH dehydrogenase subunit 5 [Necator americanus] NP_579967.2 1e-121 73% ...

  4. NCBI nr-aa BLAST: CBRC-CBRI-08-0176 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CBRI-08-0176 ref|NP_579967.2| NADH dehydrogenase subunit 5 [Necator americanus...] emb|CAD10452.2| NADH dehydrogenase subunit 5 [Necator americanus] NP_579967.2 1e-121 73% ...

  5. NCBI nr-aa BLAST: CBRC-ACAR-01-1152 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-1152 ref|YP_025913.1| NADH dehydrogenase subunit 2 [Xiphinema american...um] gb|AAQ75777.1| NADH dehydrogenase subunit 2 [Xiphinema americanum] YP_025913.1 0.062 24% ...

  6. NCBI nr-aa BLAST: CBRC-MDOM-08-0118 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-08-0118 ref|YP_025913.1| NADH dehydrogenase subunit 2 [Xiphinema american...um] gb|AAQ75777.1| NADH dehydrogenase subunit 2 [Xiphinema americanum] YP_025913.1 0.49 27% ...

  7. NCBI nr-aa BLAST: CBRC-DNOV-01-0856 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0856 ref|YP_025913.1| NADH dehydrogenase subunit 2 [Xiphinema american...um] gb|AAQ75777.1| NADH dehydrogenase subunit 2 [Xiphinema americanum] YP_025913.1 0.005 27% ...

  8. NCBI nr-aa BLAST: CBRC-MDOM-01-0308 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0308 ref|YP_001789002.1| NADH dehydrogenase subunit 2 [Phthonandria at...rilineata] gb|ACB20521.1| NADH dehydrogenase subunit 2 [Phthonandria atrilineata] YP_001789002.1 0.39 24% ...

  9. NCBI nr-aa BLAST: CBRC-OSAT-07-0033 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OSAT-07-0033 ref|YP_025744.1| NADH dehydrogenase subunit 4 [Ornithoctonus huwe...na] gb|AAP51154.2| NADH dehydrogenase subunit 4 [Ornithoctonus huwena] YP_025744.1 3.2 25% ...

  10. NCBI nr-aa BLAST: CBRC-TTRU-01-0100 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0100 ref|YP_025744.1| NADH dehydrogenase subunit 4 [Ornithoctonus huwe...na] gb|AAP51154.2| NADH dehydrogenase subunit 4 [Ornithoctonus huwena] YP_025744.1 0.012 25% ...

  11. NCBI nr-aa BLAST: CBRC-BTAU-01-1570 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-BTAU-01-1570 ref|NP_443568.1|ND2_15834 NADH dehydrogenase subunit 2 [Rondeletia lorica...ta] dbj|BAB70262.1| NADH dehydrogenase subunit 2 [Rondeletia loricata] NP_443568.1 3.9 25% ...

  12. NCBI nr-aa BLAST: CBRC-TTRU-01-0016 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0016 ref|YP_001816763.1| NADH dehydrogenase subunit 2 [Nephtys sp. 'San Juan... Island' YV-2008] gb|ACB28516.1| NADH dehydrogenase subunit 2 [Nephtys sp. 'San Juan Island' YV-2008] YP_001816763.1 5e-04 28% ...

  13. NCBI nr-aa BLAST: CBRC-TTRU-01-1049 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1049 gb|ACB46268.1| NADH dehydrogenase subunit 2 [Glossina morsitans morsitan...s] gb|ACB46269.1| NADH dehydrogenase subunit 2 [Glossina morsitans centralis] ACB46268.1 0.85 28% ...

  14. NCBI nr-aa BLAST: CBRC-ETEL-01-0669 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ETEL-01-0669 ref|YP_001382309.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infern...alis] dbj|BAF73637.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infernalis] YP_001382309.1 0.054 23% ...

  15. NCBI nr-aa BLAST: CBRC-TTRU-01-0816 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0816 ref|YP_001382309.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infern...alis] dbj|BAF73637.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infernalis] YP_001382309.1 0.014 22% ...

  16. NCBI nr-aa BLAST: CBRC-CPOR-01-1888 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CPOR-01-1888 ref|YP_001382309.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infern...alis] dbj|BAF73637.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infernalis] YP_001382309.1 0.009 22% ...

  17. NCBI nr-aa BLAST: CBRC-TTRU-01-1302 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1302 ref|YP_001382309.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infern...alis] dbj|BAF73637.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infernalis] YP_001382309.1 0.038 24% ...

  18. NCBI nr-aa BLAST: CBRC-TTRU-01-0297 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0297 ref|YP_001382309.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infern...alis] dbj|BAF73637.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infernalis] YP_001382309.1 0.17 25% ...

  19. NCBI nr-aa BLAST: CBRC-TTRU-01-0568 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0568 ref|YP_001382309.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infern...alis] dbj|BAF73637.1| NADH dehydrogenase subunit 2 [Vampyroteuthis infernalis] YP_001382309.1 0.016 21% ...

  20. NCBI nr-aa BLAST: CBRC-CINT-01-0194 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CINT-01-0194 ref|YP_514737.1| NADH dehydrogenase subunit 2 [Parakneria cameron...ensis] dbj|BAE79982.1| NADH dehydrogenase subunit 2 [Parakneria cameronensis] YP_514737.1 0.20 28% ...

  1. NCBI nr-aa BLAST: CBRC-DNOV-01-0872 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0872 ref|NP_008495.1|ND6_15043 NADH dehydrogenase subunit 6 [Artibeus jama...icensis] gb|AAD05448.1| NADH dehydrogenase subunit 6 [Artibeus jamaicensis] NP_008495.1 0.62 22% ...

  2. NCBI nr-aa BLAST: CBRC-OCUN-01-0083 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0083 ref|NP_150132.1| NADH dehydrogenase subunit 5 [Schizophyllum comm...une] gb|AAK83416.1|AF402141_20 NADH dehydrogenase subunit 5 [Schizophyllum commune] NP_150132.1 2e-88 51% ...

  3. NCBI nr-aa BLAST: CBRC-DSIM-03-0004 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DSIM-03-0004 ref|NP_150132.1| NADH dehydrogenase subunit 5 [Schizophyllum comm...une] gb|AAK83416.1|AF402141_20 NADH dehydrogenase subunit 5 [Schizophyllum commune] NP_150132.1 0.81 24% ...

  4. NCBI nr-aa BLAST: CBRC-LAFR-01-0838 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-0838 ref|NP_150124.1| NADH dehydrogenase subunit 4 [Schizophyllum comm...une] gb|AAK83408.1|AF402141_12 NADH dehydrogenase subunit 4 [Schizophyllum commune] NP_150124.1 3e-62 48% ...

  5. NCBI nr-aa BLAST: CBRC-CINT-01-0190 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CINT-01-0190 ref|YP_086875.1| NADH dehydrogenase subunit 4L [Amphisbaena schmidt...i] gb|AAT08524.1| NADH dehydrogenase subunit 4L [Amphisbaena schmidti] YP_086875.1 4.4 32% ...

  6. NCBI nr-aa BLAST: CBRC-TTRU-01-0617 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0617 ref|YP_002587081.1| NADH dehydrogenase subunit 4 [Litopenaeus sty...lirostris] gb|ACA47126.1| NADH dehydrogenase subunit 4 [Litopenaeus stylirostris] YP_002587081.1 0.24 22% ...

  7. NCBI nr-aa BLAST: CBRC-XTRO-01-2592 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-2592 ref|NP_543046.1| NADH dehydrogenase subunit 2 [Venerupis (Ruditapes) philip...pinarum] dbj|BAB83797.1| NADH dehydrogenase subunit 2 [Venerupis (Ruditapes) philippinarum] NP_543046.1 0.092 24% ...

  8. NCBI nr-aa BLAST: CBRC-ACAR-01-0325 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-0325 ref|NP_543045.1| NADH dehydrogenase subunit 1 [Venerupis (Ruditapes) philip...pinarum] dbj|BAB83796.1| NADH dehydrogenase subunit 1 [Venerupis (Ruditapes) philippinarum] NP_543045.1 6.5 26% ...

  9. NCBI nr-aa BLAST: CBRC-OLAT-26-0090 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OLAT-26-0090 ref|YP_001096004.1| NADH dehydrogenase subunit 2 [Metaseiulus occidental...is] gb|ABN45839.1| NADH dehydrogenase subunit 2 [Metaseiulus occidentalis] YP_001096004.1 6e-09 26% ...

  10. NCBI nr-aa BLAST: CBRC-MDOM-08-0012 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-08-0012 ref|YP_002889396.1| NADH dehydrogenase subunit 2 [Orussus occidental...is] gb|ACJ69694.1| NADH dehydrogenase subunit 2 [Orussus occidentalis] YP_002889396.1 0.078 26% ...

  11. NCBI nr-aa BLAST: CBRC-TTRU-01-0272 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0272 ref|YP_001096004.1| NADH dehydrogenase subunit 2 [Metaseiulus occidental...is] gb|ABN45839.1| NADH dehydrogenase subunit 2 [Metaseiulus occidentalis] YP_001096004.1 0.003 25% ...

  12. NCBI nr-aa BLAST: CBRC-TTRU-01-1208 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1208 ref|YP_002889403.1| NADH dehydrogenase subunit 5 [Orussus occidental...is] gb|ACJ69701.1| NADH dehydrogenase subunit 5 [Orussus occidentalis] YP_002889403.1 0.087 24% ...

  13. NCBI nr-aa BLAST: CBRC-OPRI-01-0198 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OPRI-01-0198 ref|YP_001096004.1| NADH dehydrogenase subunit 2 [Metaseiulus occidental...is] gb|ABN45839.1| NADH dehydrogenase subunit 2 [Metaseiulus occidentalis] YP_001096004.1 0.28 23% ...

  14. NCBI nr-aa BLAST: CBRC-ATHA-03-0003 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ATHA-03-0003 ref|YP_001096013.1| NADH dehydrogenase subunit 4 [Metaseiulus occidental...is] gb|ABN45848.1| NADH dehydrogenase subunit 4 [Metaseiulus occidentalis] YP_001096013.1 0.002 25% ...

  15. NCBI nr-aa BLAST: CBRC-ATHA-03-0003 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ATHA-03-0003 ref|YP_001096002.1| NADH dehydrogenase subunit 4 [Metaseiulus occidental...is] gb|ABN45837.1| NADH dehydrogenase subunit 4 [Metaseiulus occidentalis] YP_001096002.1 0.001 23% ...

  16. NCBI nr-aa BLAST: CBRC-VPAC-01-1521 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-VPAC-01-1521 ref|YP_002889403.1| NADH dehydrogenase subunit 5 [Orussus occidental...is] gb|ACJ69701.1| NADH dehydrogenase subunit 5 [Orussus occidentalis] YP_002889403.1 1.2 29% ...

  17. NCBI nr-aa BLAST: CBRC-TTRU-01-1108 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1108 ref|YP_002889403.1| NADH dehydrogenase subunit 5 [Orussus occidental...is] gb|ACJ69701.1| NADH dehydrogenase subunit 5 [Orussus occidentalis] YP_002889403.1 0.048 23% ...

  18. NCBI nr-aa BLAST: CBRC-TTRU-01-1062 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1062 ref|YP_002003296.1| NADH dehydrogenase subunit 4 [Unionicola foil...i] gb|ACF19644.1| NADH dehydrogenase subunit 4 [Unionicola foili] YP_002003296.1 0.006 32% ...

  19. NCBI nr-aa BLAST: CBRC-TTRU-01-1257 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1257 ref|YP_002003299.1| NADH dehydrogenase subunit 1 [Unionicola foil...i] gb|ACF19648.1| NADH dehydrogenase subunit 1 [Unionicola foili] YP_002003299.1 0.056 26% ...

  20. NCBI nr-aa BLAST: CBRC-MDOM-02-0145 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-02-0145 ref|YP_002003293.1| NADH dehydrogenase subunit 5 [Unionicola foil...i] gb|ACF19641.1| NADH dehydrogenase subunit 5 [Unionicola foili] YP_002003293.1 4e-15 28% ...

  1. NCBI nr-aa BLAST: CBRC-TTRU-01-0424 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0424 ref|YP_002003293.1| NADH dehydrogenase subunit 5 [Unionicola foil...i] gb|ACF19641.1| NADH dehydrogenase subunit 5 [Unionicola foili] YP_002003293.1 0.029 26% ...

  2. NCBI nr-aa BLAST: CBRC-MEUG-01-2775 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MEUG-01-2775 ref|YP_002003296.1| NADH dehydrogenase subunit 4 [Unionicola foil...i] gb|ACF19644.1| NADH dehydrogenase subunit 4 [Unionicola foili] YP_002003296.1 0.097 25% ...

  3. NCBI nr-aa BLAST: CBRC-MDOM-08-0012 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-08-0012 ref|YP_002003296.1| NADH dehydrogenase subunit 4 [Unionicola foil...i] gb|ACF19644.1| NADH dehydrogenase subunit 4 [Unionicola foili] YP_002003296.1 0.002 27% ...

  4. NCBI nr-aa BLAST: CBRC-PCAP-01-1017 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PCAP-01-1017 ref|NP_443464.1| NADH dehydrogenase subunit 2 [Ateleopus japonicu...s] dbj|BAB69989.1| NADH dehydrogenase subunit 2 [Ateleopus japonicus] NP_443464.1 0.093 25% ...

  5. NCBI nr-aa BLAST: CBRC-TTRU-01-1268 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1268 ref|YP_003097137.1| NADH dehydrogenase subunit 5 [Angiostrongylus... costaricensis] gb|ACT88807.1| NADH dehydrogenase subunit 5 [Angiostrongylus costaricensis] YP_003097137.1 0.002 26% ...

  6. NCBI nr-aa BLAST: CBRC-MDOM-06-0051 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-06-0051 ref|YP_003097130.1| NADH dehydrogenase subunit 2 [Angiostrongylus... costaricensis] gb|ACT88800.1| NADH dehydrogenase subunit 2 [Angiostrongylus costaricensis] YP_003097130.1 0.28 23% ...

  7. NCBI nr-aa BLAST: CBRC-TTRU-01-1195 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1195 ref|YP_003097137.1| NADH dehydrogenase subunit 5 [Angiostrongylus... costaricensis] gb|ACT88807.1| NADH dehydrogenase subunit 5 [Angiostrongylus costaricensis] YP_003097137.1 0.019 25% ...

  8. NCBI nr-aa BLAST: CBRC-MDOM-07-0060 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-07-0060 ref|YP_003097130.1| NADH dehydrogenase subunit 2 [Angiostrongylus... costaricensis] gb|ACT88800.1| NADH dehydrogenase subunit 2 [Angiostrongylus costaricensis] YP_003097130.1 0.51 22% ...

  9. NCBI nr-aa BLAST: CBRC-MDOM-01-0287 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0287 ref|YP_003097130.1| NADH dehydrogenase subunit 2 [Angiostrongylus... costaricensis] gb|ACT88800.1| NADH dehydrogenase subunit 2 [Angiostrongylus costaricensis] YP_003097130.1 0.015 22% ...

  10. NCBI nr-aa BLAST: CBRC-TTRU-01-0293 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0293 ref|YP_026098.1| NADH dehydrogenase subunit 5 [Habronattus oregon...ensis] gb|AAT02495.1| NADH dehydrogenase subunit 5 [Habronattus oregonensis] YP_026098.1 0.062 25% ...

  11. NCBI nr-aa BLAST: CBRC-PVAM-01-0785 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PVAM-01-0785 ref|YP_026099.1| NADH dehydrogenase subunit 4 [Habronattus oregon...ensis] gb|AAT02494.1| NADH dehydrogenase subunit 4 [Habronattus oregonensis] YP_026099.1 0.026 21% ...

  12. NCBI nr-aa BLAST: CBRC-SARA-01-0900 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-SARA-01-0900 ref|YP_026098.1| NADH dehydrogenase subunit 5 [Habronattus oregon...ensis] gb|AAT02495.1| NADH dehydrogenase subunit 5 [Habronattus oregonensis] YP_026098.1 0.032 22% ...

  13. NCBI nr-aa BLAST: CBRC-TTRU-01-0902 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0902 ref|YP_026098.1| NADH dehydrogenase subunit 5 [Habronattus oregon...ensis] gb|AAT02495.1| NADH dehydrogenase subunit 5 [Habronattus oregonensis] YP_026098.1 0.003 25% ...

  14. NCBI nr-aa BLAST: CBRC-TTRU-01-0490 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0490 ref|YP_026099.1| NADH dehydrogenase subunit 4 [Habronattus oregon...ensis] gb|AAT02494.1| NADH dehydrogenase subunit 4 [Habronattus oregonensis] YP_026099.1 0.13 26% ...

  15. NCBI nr-aa BLAST: CBRC-ACAR-01-0691 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-0691 ref|YP_145513.1| NADH dehydrogenase subunit 6 [Asymmetron lucayan...um] dbj|BAD72150.1| NADH dehydrogenase subunit 6 [Asymmetron lucayanum] YP_145513.1 0.77 27% ...

  16. NCBI nr-aa BLAST: CBRC-RNOR-07-0249 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-07-0249 ref|YP_001434591.1| NADH dehydrogenase subunit 2 [Asymmetron sp. ...A TK-2007] dbj|BAF76622.1| NADH dehydrogenase subunit 2 [Asymmetron sp. A TK-2007] YP_001434591.1 0.75 26% ...

  17. NCBI nr-aa BLAST: CBRC-DNOV-01-0458 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0458 ref|YP_313639.1| NADH dehydrogenase subunit 4 [Epidermophyton flo...ccosum] gb|AAW78245.1| NADH dehydrogenase subunit 4 [Epidermophyton floccosum] YP_313639.1 0.46 27% ...

  18. NCBI nr-aa BLAST: CBRC-MDOM-01-0287 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0287 ref|YP_003001967.1| NADH dehydrogenase subunit 6 [Lymantria dispa...r] gb|ACL93267.1| NADH dehydrogenase subunit 6 [Lymantria dispar] YP_003001967.1 0.044 25% ...

  19. NCBI nr-aa BLAST: CBRC-DYAK-08-0044 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DYAK-08-0044 ref|YP_001165397.1| NADH dehydrogenase subunit 4 [Phytophthora soja...e] gb|ABG54058.1| NADH dehydrogenase subunit 4 [Phytophthora sojae] YP_001165397.1 0.40 25% ...

  20. NCBI nr-aa BLAST: CBRC-DYAK-08-0034 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DYAK-08-0034 ref|YP_001165397.1| NADH dehydrogenase subunit 4 [Phytophthora soja...e] gb|ABG54058.1| NADH dehydrogenase subunit 4 [Phytophthora sojae] YP_001165397.1 0.40 25% ...

  1. NCBI nr-aa BLAST: CBRC-TGUT-37-0024 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TGUT-37-0024 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 1e-12 34% ...

  2. NCBI nr-aa BLAST: CBRC-DRER-08-0014 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-08-0014 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 1e-07 29% ...

  3. NCBI nr-aa BLAST: CBRC-DRER-04-0032 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-04-0032 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 2e-15 32% ...

  4. NCBI nr-aa BLAST: CBRC-HSAP-07-0014 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-HSAP-07-0014 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 2e-12 29% ...

  5. NCBI nr-aa BLAST: CBRC-CINT-01-0047 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CINT-01-0047 ref|NP_937906.1| NADH dehydrogenase subunit 2 [Strongyloides stercoral...is] emb|CAD90566.1| NADH dehydrogenase subunit 2 [Strongyloides stercoralis] NP_937906.1 6e-09 28% ...

  6. NCBI nr-aa BLAST: CBRC-RNOR-03-0001 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RNOR-03-0001 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 6e-04 30% ...

  7. NCBI nr-aa BLAST: CBRC-OANA-01-1383 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OANA-01-1383 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 2e-15 27% ...

  8. NCBI nr-aa BLAST: CBRC-DRER-26-0053 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-26-0053 ref|NP_937903.1| NADH dehydrogenase subunit 4 [Strongyloides stercoral...is] emb|CAD90563.1| NADH dehydrogenase subunit 4 [Strongyloides stercoralis] NP_937903.1 7e-25 30% ...

  9. NCBI nr-aa BLAST: CBRC-DNOV-01-2370 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2370 ref|NP_937907.1| NADH dehydrogenase subunit 3 [Strongyloides stercoral...is] emb|CAD90567.1| NADH dehydrogenase subunit 3 [Strongyloides stercoralis] NP_937907.1 1e-05 30% ...

  10. NCBI nr-aa BLAST: CBRC-DRER-09-0076 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-09-0076 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 5e-16 37% ...

  11. NCBI nr-aa BLAST: CBRC-XTRO-01-3943 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-3943 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 0.32 30% ...

  12. NCBI nr-aa BLAST: CBRC-DRER-26-0145 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-26-0145 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 1e-15 30% ...

  13. NCBI nr-aa BLAST: CBRC-TTRU-01-0623 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0623 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 0.007 27% ...

  14. NCBI nr-aa BLAST: CBRC-LAFR-01-2017 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-2017 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 0.86 37% ...

  15. NCBI nr-aa BLAST: CBRC-DRER-03-0034 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-03-0034 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 3e-11 30% ...

  16. NCBI nr-aa BLAST: CBRC-MMUS-23-0130 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-23-0130 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 1e-07 30% ...

  17. NCBI nr-aa BLAST: CBRC-AGAM-04-0035 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-AGAM-04-0035 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 0.009 23% ...

  18. NCBI nr-aa BLAST: CBRC-CPOR-01-0714 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CPOR-01-0714 ref|NP_937906.1| NADH dehydrogenase subunit 2 [Strongyloides stercoral...is] emb|CAD90566.1| NADH dehydrogenase subunit 2 [Strongyloides stercoralis] NP_937906.1 0.002 26% ...

  19. NCBI nr-aa BLAST: CBRC-BTAU-01-1871 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-BTAU-01-1871 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 1.4 29% ...

  20. NCBI nr-aa BLAST: CBRC-PVAM-01-0785 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PVAM-01-0785 ref|NP_937903.1| NADH dehydrogenase subunit 4 [Strongyloides stercoral...is] emb|CAD90563.1| NADH dehydrogenase subunit 4 [Strongyloides stercoralis] NP_937903.1 0.026 25% ...

  1. NCBI nr-aa BLAST: CBRC-TBEL-01-1985 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TBEL-01-1985 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 0.001 31% ...

  2. NCBI nr-aa BLAST: CBRC-DNOV-01-1062 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-1062 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 2e-05 26% ...

  3. NCBI nr-aa BLAST: CBRC-MMUS-16-0066 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUS-16-0066 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 1e-17 34% ...

  4. NCBI nr-aa BLAST: CBRC-XTRO-01-1662 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-1662 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 3e-06 29% ...

  5. NCBI nr-aa BLAST: CBRC-DRER-02-0066 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-02-0066 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 4e-24 30% ...

  6. NCBI nr-aa BLAST: CBRC-ACAR-01-1086 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-1086 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 1e-06 36% ...

  7. NCBI nr-aa BLAST: CBRC-DRER-26-0480 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-26-0480 ref|NP_937906.1| NADH dehydrogenase subunit 2 [Strongyloides stercoral...is] emb|CAD90566.1| NADH dehydrogenase subunit 2 [Strongyloides stercoralis] NP_937906.1 7e-13 29% ...

  8. NCBI nr-aa BLAST: CBRC-DNOV-01-3061 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-3061 ref|NP_937896.1| NADH dehydrogenase subunit 1 [Strongyloides stercoral...is] emb|CAD90556.1| NADH dehydrogenase subunit 1 [Strongyloides stercoralis] NP_937896.1 0.014 25% ...

  9. NCBI nr-aa BLAST: CBRC-TTRU-01-0075 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0075 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 0.038 25% ...

  10. NCBI nr-aa BLAST: CBRC-RMAC-13-0002 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-RMAC-13-0002 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 2e-18 34% ...

  11. NCBI nr-aa BLAST: CBRC-DRER-26-0182 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-26-0182 ref|NP_937906.1| NADH dehydrogenase subunit 2 [Strongyloides stercoral...is] emb|CAD90566.1| NADH dehydrogenase subunit 2 [Strongyloides stercoralis] NP_937906.1 2e-08 24% ...

  12. NCBI nr-aa BLAST: CBRC-VPAC-01-1472 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-VPAC-01-1472 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 1.4 24% ...

  13. NCBI nr-aa BLAST: CBRC-TSYR-01-0314 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TSYR-01-0314 ref|NP_937903.1| NADH dehydrogenase subunit 4 [Strongyloides stercoral...is] emb|CAD90563.1| NADH dehydrogenase subunit 4 [Strongyloides stercoralis] NP_937903.1 0.094 43% ...

  14. NCBI nr-aa BLAST: CBRC-CFAM-18-0124 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CFAM-18-0124 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 5e-19 33% ...

  15. NCBI nr-aa BLAST: CBRC-CFAM-04-0019 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CFAM-04-0019 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 5e-17 39% ...

  16. NCBI nr-aa BLAST: CBRC-GGAL-02-0006 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-GGAL-02-0006 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 2e-07 26% ...

  17. NCBI nr-aa BLAST: CBRC-DNOV-01-2370 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2370 ref|NP_937903.1| NADH dehydrogenase subunit 4 [Strongyloides stercoral...is] emb|CAD90563.1| NADH dehydrogenase subunit 4 [Strongyloides stercoralis] NP_937903.1 0.001 32% ...

  18. NCBI nr-aa BLAST: CBRC-CJAC-01-0851 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CJAC-01-0851 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 4e-09 29% ...

  19. NCBI nr-aa BLAST: CBRC-MDOM-07-0041 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-07-0041 ref|NP_937903.1| NADH dehydrogenase subunit 4 [Strongyloides stercoral...is] emb|CAD90563.1| NADH dehydrogenase subunit 4 [Strongyloides stercoralis] NP_937903.1 1e-04 28% ...

  20. NCBI nr-aa BLAST: CBRC-CINT-01-0047 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CINT-01-0047 ref|NP_937902.1| NADH dehydrogenase subunit 5 [Strongyloides stercoral...is] emb|CAD90562.1| NADH dehydrogenase subunit 5 [Strongyloides stercoralis] NP_937902.1 9e-10 27% ...