title: Kinetic and mechanistic characterization of the formyl-CoA transferase from Oxalobacter formigenes.
abstract: Oxalobacter formigenes is an obligate anaerobe that colonizes the human gastrointestinal tract and employs oxalate breakdown to generate ATP in a novel process involving the interplay of two coupled enzymes and a membrane-bound oxalate:formate antiporter. Formyl-CoA transferase is a critical enzyme in oxalate-dependent ATP synthesis and is the first Class III CoA-transferase for which a high resolution, three-dimensional structure has been determined (Ricagno, S., Jonsson, S., Richards, N., and Lindqvist, Y. (2003) EMBO J. 22, 3210-3219). We now report the first detailed kinetic characterizations of recombinant, wild type formyl-CoA transferase and a number of site-specific mutants, which suggest that catalysis proceeds via a series of anhydride intermediates. Further evidence for this mechanistic proposal is provided by the x-ray crystallographic observation of an acylenzyme intermediate that is formed when formyl-CoA transferase is incubated with oxalyl-CoA. The catalytic mechanism of formyl-CoA transferase is therefore established and is almost certainly employed by all other members of the Class III CoA-transferase family.
authors.affiliation: Department of Chemistry, University of Florida, Gainesville, Florida 32611-7200, USA.
authors.forename: Stefan
authors.initials: S
authors.lastname: Jonsson
authors.forename: Stefano
authors.initials: S
authors.lastname: Ricagno
authors.forename: Ylva
authors.initials: Y
authors.lastname: Lindqvist
authors.forename: Nigel G J
authors.initials: NG
authors.lastname: Richards
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D015256:Coenzyme A-Transferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D015394:Molecular Structure; D009154:Mutation; D020624:Oxalobacter formigenes / Q000201:enzymology*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D015256:Coenzyme A-Transferases; C064813:formyl-coenzyme A transferase