Cofactor Editing by the G-protein Metallochaperone Domain Regulates the Radical B 12 Enzyme IcmF

• Authors: Zhu Li, University of Michigan; Kenichi Kitanishi, University of Michigan; Umar T. Twahir, Emory University; Valentin Cracan, University of Michigan; Derrell Chapman, University of Michigan; Kurt Warncke, Emory University; Ruma Banerjee, University of Michigan
• Journal Title: Journal of Biological Chemistry
• Volume: Volume 292, Number 10
• Publisher: American Society for Biochemistry and Molecular Biology | 2017-03-10, Pages 3977-3987
• Type of Work: Article | Final Publisher PDF
• Publisher DOI: http://doi.org/10.1074/jbc.M117.775957
• Subjects: Biology, Molecular; Physics, General
• Keywords: adenosylcobalamin (AdoCbl); ATP; GTPase; low molecular weight G-protein; metalloprotein

Abstract

IcmF is a 5′-deoxyadenosylcobalamin (AdoCbl)-dependent enzyme that catalyzes the carbon skeleton rearrangement of isobutyryl-CoA to butyryl-CoA. It is a bifunctional protein resulting from the fusion of a G-protein chaperone with GTPase activity and the cofactor- and substrate-binding mutase domains with isomerase activity. IcmF is prone to inactivation during catalytic turnover, thus setting up its dependence on a cofactor repair system. Herein, we demonstrate that the GTPase activity of IcmF powers the ejection of the inactive cob(II)alamin cofactor and requires the presence of an acceptor protein, adenosyltransferase, for receiving it. Adenosyltransferase in turn converts cob(II)alamin to AdoCbl in the presence of ATP and a reductant. The repaired cofactor is then reloaded onto IcmF in a GTPase-gated step. The mechanistic details of cofactor loading and offloading from the AdoCbl-dependent IcmF are distinct from those of the better characterized and homologous methylmalonyl-CoA mutase/G-protein chaperone system.