Catalytic role of the conformational change in succinyl-CoA:3-oxoacid CoA transferase on binding CoA.
Catalysis by succinyl-CoA:3-oxoacid CoA transferase proceeds through a thioester intermediate in which CoA is covalently linked to the enzyme. To determine the conformation of the thioester intermediate, crystals of the pig enzyme were grown in the presence of the substrate acetoacetyl-CoA. X-ray diffraction data show the enzyme in both the free form and covalently bound to CoA via Glu305. In the complex, the protein adopts a conformation in which residues 267-275, 280-287, 357-373, and 398-477 have shifted toward Glu305, closing the enzyme around the thioester. Enzymes provide catalysis by stabilizing the transition state relative to complexes with substrates or products. In this case, the conformational change allows the enzyme to interact with parts of CoA distant from the reactive thiol while the thiol is covalently linked to the enzyme. The enzyme forms stabilizing interactions with both the nucleotide and pantoic acid portions of CoA, while the interactions with the amide groups of the pantetheine portion are poor. The results shed light on how the enzyme uses the binding energy for groups remote from the active center of CoA to destabilize atoms closer to the active center, leading to acceleration of the reaction by the enzyme.
Department of Biological Sciences, University of Calgary, 2500 University Drive NW, Calgary, Alberta, Canada. frasm@ucalgary.ca
Marie E
ME
Fraser
Koto
K
Hayakawa
William D
WD
Brown
D000214:Acyl Coenzyme A / Q000378:metabolism* / Q000494:pharmacology*; D000818:Animals; D001665:Binding Sites; D055162:Biocatalysis; D015256:Coenzyme A-Transferases / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D004795:Enzyme Stability / Q000187:drug effects; D007700:Kinetics; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects; D013552:Swine
D000214:Acyl Coenzyme A; C010667:acetoacetyl CoA; D015256:Coenzyme A-Transferases; C021460:3-ketoacid CoA-transferase