EPR evidence for nickel-substrate interaction in carbon monoxide dehydrogenase from Clostridium thermoaceticum

Ragsdale, S.W.; Ljungdahl, Lars G.; Dervartanian, D.V.

doi:10.1016/0006-291x(82)90880-4

Cited by 104 publications

(81 citation statements)

References 13 publications

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“…This signal showed broadening when evoked with 13CO but was not light sensitive [207,210]. Its line shape shows some resemblance to the CO-induced EPR signal (gxy = 2.07, gz = 2.02) in CO dehydrogenase from Clostridium thermoaceticum [155,180]. Its origin is not known.…”

Section: Binding Of Carbon Monoxide To Ni(ii)mentioning

confidence: 95%

Nickel hydrogenases: in search of the active site

Albracht

1994

Biochimica et Biophysica Acta (BBA) - Bioenergetics

470

479

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Section: Binding Of Carbon Monoxide To Ni(ii)mentioning

confidence: 95%

Nickel hydrogenases: in search of the active site

Albracht

1994

Biochimica et Biophysica Acta (BBA) - Bioenergetics

470

479

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“…In CDH from anaerobic microorganisms, at least one putative [4Fe-4S] cubane structure has been detected by EPR spectroscopy [13,[14][15][16][17]331. It is likely that this cluster with an Em around -400 mV functions as an electron acceptor in the oxidation of CO [7-101.…”

Section: Disc Ussl Onmentioning

confidence: 99%

EPR characterization of a high‐spin system in carbon monoxide dehydrogenase from Methanothrix soehngenii

Jetten

Pierik

Hagen

1991

European Journal of Biochemistry

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Carbon monoxide dehydrogenase and methyl-coenzyme M reductase were purified from 61Ni-enriched and natural-abundance nickel-grown cells of the methanogenic archae Methanothrix soehngenii. The nickel-EPR signal from cofactor F-430 in methyl-CoM reductase was of substoichiometric intensity and exhibited near-axial symmetry with g = 2.153, 2.221 and resolved porphinoid nitrogen superhyperfine splittings of z 1 mT. In the spectrum from 61Ni-enriched enzyme a wellresolved parallel I = 3/2 nickel hyperfine splitting was observed, All = 4.4 mT. From a computer simulation of this spectrum the final enrichment in 61Ni was estimated to be 69%, while the original enrichment of the nickel metal was 87%. Carbon monoxide dehydrogenase isolated from the same batch exhibited four different EPR spectra. However, in none of these signals could any splitting or broadening from "Ni be detected. Also, the characteristic g = 2.08 EPR signal found in some other carbon monoxide dehydrogenases and ascribed to a Ni-Fe-C complex, was never observed by us under any conditions of detection (4 to 100 K) and incubation in the presence of ferricyanide, dithionite, CO, coenzyme A, or acetyl-coenzyme A. Novel, high-spin EPR was found in the oxidized enzyme with effective g-values at g = 14.5, 9.6, 5.5, 4.6, 4.2, 3.8. The lines at g = 14.5 and 5.5 were tentatively ascribed to an S = 912 system (~0 . 3 spinslap) with rhombicity EID = 0.047 and D < 0.The other signals were assigned to an S = 512 system (0.1 spinslag) with E / D = 0.27. Both sets of signals disappear upon reduction with Em, 7 . 5 = -280 mV. With a very similar reduction potential, Em,,,5 = -261 mV, an S = l j 2 signal (0.1 spinslap) appears with the unusual g-tensor 2.005, 1.894, 1.733. Upon further lowering of the potential the putative double cubane signal also appears. At a potential E z -320 mV the double cubane is only reduced by a few percent and this allows the detection of individual cubane EPR not subjected to dipolar interaction; a single spectral component is observed with g-tensor 2.048, 1.943, 1.894.Carbon monoxide dehydrogenase (CDH) is the central enzyme in the Wood pathway for autotrophic acetylcoenzyme A (CoASAc) synthesis by homoacetogenic bacteria [I -31. In these bacteria the enzyme catalyzes several reactions, the most important being the reduction of C 0 2 to CO and the condensation of a methyl group bound to CDH with CO and CoASH to CoASAc [4 -61. CDH also plays an important role in acetate degradation by methanogenic archae, where the enzyme catalyzes the oxidation of CO to COz and the cleavage of CoASAc to a methyl, carbonyl and CoASH moiety [7-101. Although C D H activity has opposite functions in the metabolisms of these two microbial groups, on the molecular level the enzymes have several properties in common. CDH constitutes up to 2 -5% of the soluble protein and this reflects its metabolic importance. The enzyme has a tetrameric structure ( M , Y )~, but it can be part of a complex [4, lo]. The genes for the subunits are clustered in a opcronlike struct...

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“…Most other hyperthermophiles, the majority anaerobes, were isolated by Karl Stetter (141) in the 1980s and 1990s. The organism with the highest growth temperature (122 • C) that is presently known is Methanopyrus kandleri, a strictly anaerobic methanogenic archaeon. No one in the 1960s would have believed that life is possible, and might even have evolved, at temperatures >100…”

Section: Fifty Years Ago: What Was Not Yet Knownmentioning

confidence: 99%

“…With partially purified (14-fold) enzyme of M. thermoacetica, Harland Wood's lab subsequently showed, in 1980, that nickel and CO dehydrogenase activity cochromatograph in polyacrylamide gels (46). Final evidence that CO dehydrogenase is a nickel enzyme was then provided at the end of 1982, when the enzyme was purified to apparent homogeneity (40,122). The heterodimeric enzyme complex contained two to three nickel atoms per mol.…”

Section: Nickel In Carbon Monoxide Dehydrogenase (1979)mentioning

confidence: 99%

My Lifelong Passion for Biochemistry and Anaerobic Microorganisms

Thauer

2015

Annu. Rev. Microbiol.

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Early parental influence led me first to medical school, but after developing a passion for biochemistry and sensing the need for a deeper foundation, I changed to chemistry. During breaks between semesters, I worked in various biochemistry labs to acquire a feeling for the different areas of investigation. The scientific puzzle that fascinated me most was the metabolism of the anaerobic bacterium Clostridium kluyveri, which I took on in 1965 in Karl Decker's lab in Freiburg, Germany. I quickly realized that little was known about the biochemistry of strict anaerobes such as clostridia, methanogens, acetogens, and sulfate-reducing bacteria and that these were ideal model organisms to study fundamental questions of energy conservation, CO 2 fixation, and the evolution of metabolic pathways. My passion for anaerobes was born then and is unabated even after 50 years of study.

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EPR evidence for nickel-substrate interaction in carbon monoxide dehydrogenase from Clostridium thermoaceticum

Cited by 104 publications

References 13 publications

Nickel hydrogenases: in search of the active site

Nickel hydrogenases: in search of the active site

EPR characterization of a high‐spin system in carbon monoxide dehydrogenase from Methanothrix soehngenii

My Lifelong Passion for Biochemistry and Anaerobic Microorganisms

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