title: Crystallographic trapping of the glutamyl-CoA thioester intermediate of family I CoA transferases.
abstract: Coenzyme A transferases are involved in a broad range of biochemical processes in both prokaryotes and eukaryotes, and exhibit a diverse range of substrate specificities. The YdiF protein from Escherichia coli O157:H7 is an acyl-CoA transferase of unknown physiological function, and belongs to a large sequence family of CoA transferases, present in bacteria to humans, which utilize oxoacids as acceptors. In vitro measurements showed that YdiF displays enzymatic activity with short-chain acyl-CoAs. The crystal structures of YdiF and its complex with CoA, the first co-crystal structure for any Family I CoA transferase, have been determined and refined at 1.9 and 2.0 A resolution, respectively. YdiF is organized into tetramers, with each monomer having an open alpha/beta structure characteristic of Family I CoA transferases. Co-crystallization of YdiF with a variety of CoA thioesters in the absence of acceptor carboxylic acid resulted in trapping a covalent gamma-glutamyl-CoA thioester intermediate. The CoA binds within a well defined pocket at the N- and C-terminal domain interface, but makes contact only with the C-terminal domain. The structure of the YdiF complex provides a basis for understanding the different catalytic steps in the reaction of Family I CoA transferases.
authors.affiliation: Department of Biochemistry, McGill University, Quebec, Canada.
authors.forename: Erumbi S
authors.initials: ES
authors.lastname: Rangarajan
authors.forename: Yunge
authors.initials: Y
authors.lastname: Li
authors.forename: Eunice
authors.initials: E
authors.lastname: Ajamian
authors.forename: Pietro
authors.initials: P
authors.lastname: Iannuzzi
authors.forename: Stephanie D
authors.initials: SD
authors.lastname: Kernaghan
authors.forename: Marie E
authors.initials: ME
authors.lastname: Fraser
authors.forename: Miroslaw
authors.initials: M
authors.lastname: Cygler
authors.forename: Allan
authors.initials: A
authors.lastname: Matte
mesh_terms: D001665:Binding Sites; D002264:Carboxylic Acids / Q000737:chemistry; D002384:Catalysis; D020134:Catalytic Domain; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D003065:Coenzyme A / Q000737:chemistry*; D015256:Coenzyme A-Transferases / Q000737:chemistry* / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D004952:Esters / Q000737:chemistry*; D018698:Glutamic Acid / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry; D013058:Mass Spectrometry; D008956:Models, Chemical; D008958:Models, Molecular; D008968:Molecular Conformation; D011487:Protein Conformation; D017510:Protein Folding; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
chemical_list: D002264:Carboxylic Acids; D029968:Escherichia coli Proteins; D004952:Esters; D018698:Glutamic Acid; D006639:Histidine; D015256:Coenzyme A-Transferases; C507992:YdiF protein, E coli; D003065:Coenzyme A