REAL

Characterization of a novel acylaminoacyl peptidase with hexameric structure and endopeptidase activity

Szeltner, Zoltán and Kiss, András and Domokos, Klarissza and Harmat, Veronika and Náray-Szabó, Gábor and Polgár, László (2009) Characterization of a novel acylaminoacyl peptidase with hexameric structure and endopeptidase activity. Biochimica et Biophysica Acta, 1794 (8). pp. 1204-1210. ISSN 1570-9639

[img] PDF
BBA.pdf
Restricted to Registered users only

Download (312Kb)

Abstract

We have overexpressed in E. coli, purified and investigated the kinetic, thermodynamic and biophysical properties of an acylaminoacyl peptidase (AAP), from the thermophile Pyrococcus horikoshii (PhAAP). It was shown that the electrostatic environment of the catalytic site of PhAAP substantially influenced the pH dependence of the specificity rate constant (kcat/Km). However, 0.3 M NaCl, which depressed the electrostatic effects, simplified the complex pH-rate profile. The rate of formation of the enzyme–substrate complex (k1) was obtained from a non-linear Arrhenius plot. The lack of substrate leaving group effects indicated that k1 is the rate determining step in the catalysis. DSC and CD measurements demonstrated that PhAAP displayed a stable structure in the catalytically competent pH range. It was shown that PhAAP is not just an acylaminoacyl peptidase, but it also has an endopeptidase activity and so differs from the mammalian AAPs. Size exclusion chromatography with PhAAP revealed a hexameric structure, which is unique among the known members of the prolyl oligopeptidase family that includes AAPs and suggests that its cellular function may be different from that of the dimeric AAP also found in the same organism.

Item Type: Article
Subjects: Q Science / természettudomány > QD Chemistry / kémia > QD05 Crystallography / kristálytan
Depositing User: Prof Gábor Náray-Szabó
Date Deposited: 06 Feb 2012 14:34
Last Modified: 08 Oct 2012 10:49
URI: http://real.mtak.hu/id/eprint/2821

Actions (login required)

View Item View Item