Analysis of conserved residues of the human puromycin-sensitive aminopeptidase

被引：13

作者：

Thompson, MW ^{[1
]}

Hersh, LB ^{[1
]}

机构：

[1] Univ Kentucky, Dept Cellular & Mol Biochem, Lexington, KY 40536 USA

来源：

PEPTIDES | 2003年 / 24卷 / 09期

关键词：

aminopeptidase; substrate binding; catalysis; mechanism; mutagenesis; catalytic residues; bestatin; puromycin;

D O I：

10.1016/j.peptides.2003.07.012

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The puromycin-sensitive aminopeptidase (ApPS) is a zinc metallopeptidase involved in the degradation of neuropeptides. Putative catalytic residues of the enzyme, Cys146, Glu338, and Lys396 were mutated, and the resultant mutant enzymes characterized. ApPS C146S exhibited normal catalytic activity, ApPS E338A exhibited decreased substrate binding, and ApPS K396I exhibited decreases in both substrate binding and catalysis. ApPS K396I and ApPS Y394F were analyzed with respect to transition state inhibitor binding. No effect was seen with the K396I mutation, but ApPS Y394F exhibited a 3.3-fold lower affinity for RB-3014, a transition state inhibitor, indicating that Tyr394 is involved in transition state stabilization. (C) 2003 Elsevier Inc. All rights reserved.

引用

页码：1359 / 1365

页数：7

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