Functional role of a non-active site residue Trp23 on the enzyme activity of Escherichia coli thioesterase I/protease I/lysophospholipase L1

Li Chiun Lee, Yi Li Chou, Hong Hwa Chen, Ya Lin Lee, Jei Fu Shaw

研究成果: Article同行評審

12 引文 斯高帕斯(Scopus)

摘要

Escherichia coli possesses a versatile protein with the enzyme activities of thioesterase I, protease I, and lysophospholipase L1. The protein is dubbed as TAP according to the chronological order of gene discovery (TesA/ApeA/PldC). Our previous studies showed that TAP comprises the catalytic triad Ser10, Asp154, and His157 as a charge relay system, as well as Gly44 and Asn73 residues devoted to oxyanion hole stabilization. Geometrically, about 10 Å away from the enzyme catalytic cleft, Trp23 showed a stronger resonance shift than the backbone amide resonance observed in the nuclear magnetic resonance (NMR) analyses. In the present work, we conducted site-directed mutagenesis to change Trp into alanine (Ala), phenylalanine (Phe), or tyrosine (Tyr) to unveil the role of the Trp23 indole ring. Biochemical analyses of the mutant enzymes in combination with TAP's three-dimensional structures suggest that by interlinking the residues participating in this catalytic machinery, Trp23 could effectively influence substrate binding and the following turnover number. Moreover, it may serve as a contributor to both H-bond and aromatic-aromatic interaction in maintaining the cross-link within the interweaving framework of protein.

原文English
頁(從 - 到)1467-1473
頁數7
期刊Biochimica et Biophysica Acta - Proteins and Proteomics
1794
發行號10
DOIs
出版狀態Published - 2009 十月 1

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biophysics
  • Biochemistry
  • Molecular Biology

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