Binding of carbon disulfide to the site of acetyl-CoA synthesis by the nickel-iron-sulfur protein, carbon monoxide dehydrogenase, from Clostridium thermoaceticum.

Abstract

Carbon monoxide dehydrogenase (CODH) is a key enzyme in the pathway of carbon monoxide and carbon dioxide fixation by anaerobic bacteria. It performs the oxidation of CO to CO2, the reduction of CO2 to CO, and the synthesis of acetyl-CoA from a methylated corrinoid/iron-sulfur protein, CO, and CoA. These reactions occur at metal centers on CODH and involve metal-carbon bond formation and transformation. There are three iron-containing centers that play distinct roles in CODH: Centers A, B, and C. Center A is the site of synthesis of acetyl-CoA and catalyzes an exchange reaction between CO and acetyl-CoA. Center C is the site of CO oxidation and CO2 reduction. In the work described here, inhibition of CODH by carbon disulfide was studied. CS2 was found to serve as a probe of the interaction of CODH with CO at Center A. EPR spectroscopic and steady-state kinetic studies demonstrated that CS2 mimics the binding of CO to the nickel/iron-sulfur cluster at Center A; however, CS2 itself does not undergo oxidation-reduction and does not appear to bind to Center C as does CO. In the isotope exchange reaction between acetyl-CoA and CO, CS2 was found to be a competitive inhibitor with respect to CO (Ki = 0.47 mM) and a mixed inhibitor with respect to acetyl-CoA (Ki1 = 0.30 and Ki2 = 1.1 mM). The reaction of dithionite-reduced CODH with CS2 resulted in an EPR spectrum with g values of 2,200, 2,087, and 2,017.(ABSTRACT TRUNCATED AT 250 WORDS)