pH dependence studies provide insight into the structure and mechanism of thimet oligopeptidase (EC 3.4.24.15).

Abstract:

:Thimet oligopeptidase (EC 3.4.24.15; TOP) is a Zn(II) endopeptidase implicated in physiological regulation of processes involving neuropeptides. The present study clarifies the active site structure and mechanism of catalysis of TOP. The enzyme exhibited a bell-shaped pH dependence of activity having an acidic limb due to a protonation event with a pK(a) of 5.7 and a basic limb with pK(a) of 8.8. The acidic limb can be attributed to protonation of a residue affecting k(cat) while the alkaline limb may be due to conformational change. Mutation of Tyr612 to Phe resulted in more than 400-fold decrease in activity. This result, supported by modeling studies, implicates Tyr612 in transition state stabilization analogous to the role of His231 of thermolysin.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Sigman JA,Edwards SR,Pabon A,Glucksman MJ,Wolfson AJ

doi

10.1016/s0014-5793(03)00548-9

keywords:

subject

Has Abstract

pub_date

2003-06-19 00:00:00

pages

224-8

issue

2-3

eissn

0014-5793

issn

1873-3468

pii

S0014579303005489

journal_volume

545

pub_type

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