role of motif iii in catalysis by acetyl-coa synthetase第三主题由乙酰辅酶a合成酶催化的作用.pdf

Hindawi Publishing Corporation Archaea Volume 2012, Article ID 509579, 8 pages doi:10.1155/2012/509579 Research Article Role of Motif III in Catalysis by Acetyl-CoA Synthetase Cheryl Ingram-Smith, Jerry L. Thurman Jr., Karen Zimowski, and Kerry S. Smith Department of Genetics and Biochemistry, Clemson University, Clemson, SC 29634-0318, USA Correspondence should be addressed to Kerry S. Smith, kssmith@ Received 31 May 2012; Revised 12 July 2012; Accepted 30 July 2012 Academic Editor: Herman van Tilbeurgh Copyright © 2012 Cheryl Ingram-Smith et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The acyl-adenylate-forming enzyme superfamily, consisting of acyl- and aryl-CoA synthetases, the adenylation domain of the nonribosomal peptide synthetases, and luciferase, has three signature motifs (I–III) and ten conserved core motifs (A1–A10), some of which overlap the signature motifs. The consensus sequence for signature motif III (core motif A7) in acetyl-CoA synthetase is Y-X-S/T/A-G-D, with an invariant fifth position, highly conserved first and fourth positions, and variable second and third positions. Kinetic studies of enzyme variants revealed that an alteration at any position resulted in a strong decrease in the catalytic rate, although the most deleterious effects were observed when the first or fifth positions were changed. Structural modeling suggests that the highly conserved Tyr in the first position plays a key role in active site architecture through interaction with a highly conserved active-site Gln, and the invariant Asp in the fifth position plays a critical role in ATP binding and catalysis