Electron Transfer Ability from NADH to Menaquinone and from NADPH to Oxygen of Type II NADH Dehydrogenase of<i>Corynebacterium glutamicum</i>

Nantapong, Nawarat; Otofuji, Asuka; Migita, Catharina T.; Adachi, Osao; Toyama, Hirohide; Matsushita, Kazunobu

doi:10.1271/bbb.69.149

Cited by 37 publications

(30 citation statements)

References 38 publications

(58 reference statements)

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“…We also examined NADPH as an electron donor; however, Mtb NDH-2 did not show any detectable NADPH-Q oxidoreductase activity within a pH range of 4.0 -9.0. In addition, we did not detect electron transfer activity to oxygen (NAD(P)H oxidase activity), unlike NDH-2 from S. cerevisiae and C. glutamicum, as reported earlier (35).…”

Section: Nd 29supporting

confidence: 85%

“…The activity in the absence of inhibitor (ϳ15 unit mg Ϫ1 ) was normalized to 100%, and the remaining activities in the presence of inhibitors were plotted as a function of inhibitor concentration. C. glutamicum (35), and Trypanosoma brucei (38) have been reported to reduce oxygen by a single electron to produce harmful reactive oxygen species. Although it remains unknown whether reactive oxygen species produced by other NDH-2s play any physiological role, electron transfer without electron leak to oxygen, as seen in Mtb NDH-2, ensures efficient respiratory activity and reduces exogenous oxidative stress.…”

Section: Discussionmentioning

confidence: 99%

“…Among a number of different types of detergents tested, only potassium cholate was able to solubilize Mtb NDH-2 from the membranes without losing enzymatic activity. Mtb NDH-2 lost activity immediately after the addition of Triton X-100 and dodecyl-␤-D-maltoside, which have been successfully used for purification of NDH-2 from other organisms, such as E. coli (3,4) and C. glutamicum (7,35). Incubation of Mtb cytoplasmic membranes with these detergents or in alkaline buffer resulted in immediate loss of the enzymatic activity of the native NDH-2.…”

Section: Spectroscopic and Enzymatic Properties Of The Purified Mtb Nmentioning

confidence: 99%

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Steady-state Kinetics and Inhibitory Action of Antitubercular Phenothiazines on Mycobacterium tuberculosis Type-II NADH-Menaquinone Oxidoreductase (NDH-2)

Yano

Weinstein

et al. 2006

Journal of Biological Chemistry

127

133

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Type-II NADH-menaquinone oxidoreductase (NDH-2) is an essential respiratory enzyme of the pathogenic bacterium Mycobacterium tuberculosis (Mtb) that plays a pivotal role in its growth. In the present study, we expressed and purified highly active Mtb NDH-2 using a Mycobacterium smegmatis expression system, and the steady-state kinetics and inhibitory actions of phenothiazines were characterized. Purified NDH-2 contains a non-covalently bound flavin adenine dinucleotide cofactor and oxidizes NADH with quinones but does not react with either NADPH or oxygen. Ubiquinone-2 (Q2) and decylubiquinone showed high electron-accepting activity, and the steady-state kinetics and the NADH-Q2 oxidoreductase reaction were found to operate by a ping-pong reaction mechanism. Phenothiazine analogues, trifluoperazine, Compound 1, and Compound 2 inhibit the NADH-Q2 reductase activity with IC 50 ‫؍‬ 12, 11, and 13 M, respectively. Trifluoperazine inhibition is non-competitive for NADH, whereas the inhibition kinetics is found to be uncompetitive in terms of Q2.The Gram-positive bacterium Mycobacterium tuberculosis (Mtb) 3 causes tuberculosis, one of the leading causes of morbidity and mortality in the world. Each year nine million active cases of the disease are diagnosed, accounting for three million deaths. Multidrug-resistant tuberculosis and the existence of "persistent" organisms that are tolerant to antibiotics exacerbate the problem, for which more effective and efficient treatments need to be urgently developed. Mtb is traditionally considered an obligate aerobe, yet during the normal course of events in the infectious cycle, the bacillus is able to survive in conditions of low oxygen and nutrient concentrations, such as those postulated to exist within granulomas. Mtb adapts its metabolic activity, cellular transcription, and protein expression accordingly (1). It is therefore of great importance to understand how Mtb generates ATP under a variety of environmental conditions. Type-II NADH-dehydrogenase (NDH-2) is a critical enzyme in the life cycle of Mtb. The enzyme has been purified from Saccharomyces cerevisiae (2), Escherichia coli (3, 4), Bacillus subtilis (5), Methyloccocus capsulatus (6), Corynebacterium glutamicum (7, 8), Acidianus ambicalens (9, 10), and Sulfolobus metallicus (11) and is, in general, composed of a single polypeptide chain, which contains a flavin as a sole cofactor. It is noteworthy that this enzyme is not found in mitochondria. The essential role of NDH-2 in Mtb is supported by extensive evidence from biochemical (12) and transcriptional studies (13), gene deletion analysis, investigation of bacterial growth in various media and under various culture conditions, and animal experiments (12). Mtb contains two copies of ndh genes (ndh and ndhA). The Mtb NDH-2 and NDH-2A share 67% sequence identity, and the genes are separated by 17 kb. Mtb NDH-2 is highly homologous to those of Mycobacterium leprae and Mycobacterium smegmatis with 91 and 81% amino acid sequence identity, respectively. A strain of Mtb...

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Section: Nd 29supporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Section: Spectroscopic and Enzymatic Properties Of The Purified Mtb Nmentioning

confidence: 99%

See 1 more Smart Citation

Steady-state Kinetics and Inhibitory Action of Antitubercular Phenothiazines on Mycobacterium tuberculosis Type-II NADH-Menaquinone Oxidoreductase (NDH-2)

Yano

Weinstein

et al. 2006

Journal of Biological Chemistry

127

133

View full text Add to dashboard Cite

show abstract

“…4). Previous studies with purified NDH-II have shown different activities at different pHs (56). At a lower pH (pH 5.5), NDH-II activities with NADPH as a cofactor were higher in crude extracts of GSM1 (0.06 Ϯ 0.01 U/mg) than in crude extracts of the WT strain (0.01 Ϯ 0.00 U/mg).…”

Section: Design Of a C Glutamicum Strain With Atp-neutral Glycolysismentioning

confidence: 84%

Metabolic Engineering of an ATP-Neutral Embden-Meyerhof-Parnas Pathway in Corynebacterium glutamicum: Growth Restoration by an Adaptive Point Mutation in NADH Dehydrogenase

Reddy

Lindner

Wendisch

2015

Appl Environ Microbiol

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Corynebacterium glutamicum uses the Embden-Meyerhof-Parnas pathway of glycolysis and gains 2 mol of ATP per mol of glucose by substrate-level phosphorylation (SLP). To engineer glycolysis without net ATP formation by SLP, endogenous phosphorylating NAD-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was replaced by nonphosphorylating NADPdependent glyceraldehyde-3-phosphate dehydrogenase (GapN) from Clostridium acetobutylicum, which irreversibly converts glyceraldehyde-3-phosphate (GAP) to 3-phosphoglycerate (3-PG) without generating ATP. As shown recently (S. Takeno, R. Murata, R. Kobayashi, S. Mitsuhashi, and M. Ikeda, Appl Environ Microbiol 76:7154 -7160, 2010, http://dx.doi.org/10.1128/AEM.01464-10), this ATP-neutral, NADPH-generating glycolytic pathway did not allow for the growth of Corynebacterium glutamicum with glucose as the sole carbon source unless hitherto unknown suppressor mutations occurred; however, these mutations were not disclosed. In the present study, a suppressor mutation was identified, and it was shown that heterologous expression of udhA encoding soluble transhydrogenase from Escherichia coli partly restored growth, suggesting that growth was inhibited by NADPH accumulation. Moreover, genome sequence analysis of second-site suppressor mutants that were able to grow faster with glucose revealed a single point mutation in the gene of non-proton-pumping NADH:ubiquinone oxidoreductase (NDH-II) leading to the amino acid change D213G, which was shared by these suppressor mutants. Since related NDH-II enzymes accepting NADPH as the substrate possess asparagine or glutamine residues at this position, D213G, D213N, and D213Q variants of C. glutamicum NDH-II were constructed and were shown to oxidize NADPH in addition to NADH. Taking these findings together, ATP-neutral glycolysis by the replacement of endogenous NAD-dependent GAPDH with NADP-dependent GapN became possible via oxidation of NADPH formed in this pathway by mutant NADPH-accepting NDH-II D213G and thus by coupling to electron transport phosphorylation (ETP).A TP generation occurs naturally by substrate-level phosphorylation (SLP), electron transport phosphorylation (ETP), photophosphorylation, and decarboxylation phosphorylation. The Embden-Meyerhof-Parnas (EMP) pathway of glycolysis supplies ATP by SLP in the absence of terminal electron acceptors or anaerobic conditions, as well as supplying direct precursors for biomass formation (1). Bacteria and archaea possess one or more of multiple biologically feasible routes for glucose catabolism, such as the EMP pathway, the Entner-Doudoroff (ED) pathway, and the phosphoketolase pathway, which yield zero to 3 ATP molecules per glucose molecule and exist in different variants. In natural habitats, a trade-off between the rate and the yield of ATP production is observed for heterotrophic organisms; faster but less efficient ATP production may be advantageous under conditions characterized by a low abundance of growth substrates (2, 3). For example, Zymomonas mobilis ferments suga...

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“…NDH-2-type dehydrogenases also acting as an NADH:MK oxidoreductase are also ubiquitous in bacteria but are simpler and smaller in structure than NDH-1. The NDH-2-type dehydrogenases lack proton pumping activity (Nantapong et al, 2005). This type of dehydrogenase might be crucial for substrate oxidation under relatively transmembrane potential-rich conditions.…”

Section: Menaquinone In the Prokaryotic Respiratory Chainmentioning

confidence: 99%

Menaquinone as Well as Ubiquinone as a Crucial Component in the Escherichia coli Respiratory Chain

Fujimoto¹,

Kosaka²,

Yamada³

2012

Chemical Biology

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Electron Transfer Ability from NADH to Menaquinone and from NADPH to Oxygen of Type II NADH Dehydrogenase ofCorynebacterium glutamicum

Cited by 37 publications

References 38 publications

Steady-state Kinetics and Inhibitory Action of Antitubercular Phenothiazines on Mycobacterium tuberculosis Type-II NADH-Menaquinone Oxidoreductase (NDH-2)

Steady-state Kinetics and Inhibitory Action of Antitubercular Phenothiazines on Mycobacterium tuberculosis Type-II NADH-Menaquinone Oxidoreductase (NDH-2)

Metabolic Engineering of an ATP-Neutral Embden-Meyerhof-Parnas Pathway in Corynebacterium glutamicum: Growth Restoration by an Adaptive Point Mutation in NADH Dehydrogenase

Menaquinone as Well as Ubiquinone as a Crucial Component in the Escherichia coli Respiratory Chain

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