Characterization of the Ni-Fe-C Complex Formed by Reaction of Carbon Monoxide with the Carbon Monoxide Dehydrogenase from Clostridium thermoaceticum by Q-Band ENDOR

Chaoliang Fan, Carol M. Gorst*, Stephen W. Ragsdale, Brian M. Hoffman

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

91 Scopus citations

Abstract

Q-Band EN DOR studies on carbon monoxide dehydrogenase (CODH) from the acetogenic bacterium Clostridium thermoaceticum provided unambiguous evidence that the reaction of CO with CODH produces a novel metal center that includes at least one nickel, at least three iron sites, and the carbon of one CO. The 57Fe hyperfine couplings determined by ENDOR are similar to the values used in simulation of the Mössbauer spectra [Lindahl et al. (1989) J. Biol. Chem. 265, 3880–3888]. EPR simulation using these AFe values is equally good for a 4Fe or a 3 Fe center. The 13C ENDOR data are consistent with the binding of a carbon atom to either the Ni or the Fe component of the spin-coupled cluster. The 13C hyperfine couplings are similar to those determined earlier for the CO-bound form of the H cluster of the Clostridium pasteurianum hydrogenase, proposed to be the active site of hydrogen activation [Telser et al. (1987) J. Biol. Chem. 262, 6589–5694]. The 61 Ni ENDOR data are the first nickel ENDOR recorded for an enzyme. The EPR simulation using the ENDOR-derived hyperfine values for 61Ni is consistent with a single nickel site in the Ni-Fe-C complex. On the basis of our results and the Mössbauer data [Lindahl et al. (1989) J. Biol. Chem. 265, 3880–3888], we propose the stoichiometry of the components of the Ni-Fe-C complex to be Ni1Fe3_4S≥4C1, with four acid-labile sulfides.

Original languageEnglish (US)
Pages (from-to)431-435
Number of pages5
JournalBiochemistry
Volume30
Issue number2
DOIs
StatePublished - May 1 1991

ASJC Scopus subject areas

  • Biochemistry

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