Interaction of ferredoxin with carbon monoxide dehydrogenase from Clostridium thermoaceticum.
Acetogenic bacteria, as determined with Clostridium thermoaceticum, synthesize acetate by the acetyl-CoA pathway which involves the reduction of CO2 to a methyl group and then combination of the methyl with CoA and a carbonyl group formed from CO or CO2 (Wood, H.G., Ragsdale, S.W., and Pezacka, E. (1986) Trends Biochem. Sci. 11, 14-18). Carbon monoxide dehydrogenase (CODH), the key enzyme in this pathway not only catalyzes the oxidation of CO to CO2 but also the final step, the synthesis of acetyl-CoA from a methyl group, CO, and CoA. Previously, it has been shown that ferredoxin can stimulate exchange of CO with CH3 14COSCoA (Ragsdale, S.W., and Wood, H.G. (1985) J. Biol. Chem. 260, 3970-3977). In the present study, it has been observed that ferredoxin and CODH can form an electrostatically stabilized complex. In order to identify the ferredoxin binding region on CODH, the ferredoxin and CODH were cross-linked by using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. The cross-linked CODH-ferredoxin adduct was enzymatically as active as the uncross-linked complex. The native CODH and cross-linked CODH-ferredoxin complex were subjected to cyanogen bromide cleavage. By comparison of the high-performance liquid chromatography peptide profiles, it was observed that the mobility of at least one peptide is altered in the CODH-ferredoxin cross-linked complex. The peptide was identified with residues 229-239 of the alpha-subunit of CODH.
Shanmugasundaram
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Department of Biochemistry, Case Western Reserve University, Cleveland, Ohio 44106.
Wood
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HG
D000445:Aldehyde Oxidoreductases / Q000378:metabolism*; D000595:Amino Acid Sequence; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D003013:Clostridium / Q000201:enzymology*; D003432:Cross-Linking Reagents; D003488:Cyanogen Bromide; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry; D005288:Ferredoxins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D009994:Osmolar Concentration; D010084:Oxidation-Reduction
D003432:Cross-Linking Reagents; D005288:Ferredoxins; D009097:Multienzyme Complexes; D000445:Aldehyde Oxidoreductases; C019823:carbon monoxide dehydrogenase; D003488:Cyanogen Bromide; D005022:Ethyldimethylaminopropyl Carbodiimide