A Biochemical Nickel(I) State Supports Nucleophilic Alkyl Addition: A Roadmap for Methyl Reactivity in Acetyl Coenzyme A Synthase

Abstract

Nickel-containing enzymes such as methyl coenzyme M reductase (MCR) and carbon monoxide dehydrogenase/acetyl coenzyme A synthase (CODH/ACS) play a critical role in global energy conversion reactions, with significant contributions to carbon-centered processes. These enzymes are implied to cycle through a series of nickel-based organometallic intermediates during catalysis, though identification of these intermediates remains challenging. In this work, we have developed and characterized a nickel-containing metalloprotein that models the methyl-bound organometallic intermediates proposed in the native enzymes. Using a nickel(I)-substituted azurin mutant, we demonstrate that alkyl binding occurs via nucleophilic addition of methyl iodide as a methyl donor. The paramagnetic Ni ^III-CH ₃ species initially generated can be rapidly reduced to a high-spin Ni ^II-CH ₃ species in the presence of exogenous reducing agent, following a reaction sequence analogous to that proposed for ACS. These two distinct bioorganometallic species have been characterized by optical, EPR, XAS, and MCD spectroscopy, and the overall mechanism describing methyl reactivity with nickel azurin has been quantitatively modeled using global kinetic simulations. A comparison between the nickel azurin protein system and existing ACS model compounds is presented. Ni ^III-CH ₃ Az is only the second example of two-electron addition of methyl iodide to a Ni ^I center to give an isolable species and the first to be formed in a biologically relevant system. These results highlight the divergent reactivity of nickel across the two intermediates, with implications for likely reaction mechanisms and catalytically relevant states in the native ACS enzyme.

Abstract

A bioorganometallic model for acetyl coenzyme A synthase has been developed. This model protein is able to bind a cationic methyl group via direct addition to the nickel(I) center. The resultant nickel(III)-methyl species has been characterized via optical and electron paramagnetic resonance spectroscopy, and the reduced nickel(II)-methyl state has been characterized using magnetic circular dichroism and X-ray spectroscopy. Implications for further reactivity with CO are gleaned from electronic structure analysis of the nickel-methyl species.