The active site residue V266 of chlamydial HtrA is critical for substrate binding during both in vitro and in vivo conditions

S Gloeckl; J D A Tyndall; S H Stansfield; Peter Timms; W M Huston

doi:10.1159/000336312

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The active site residue V266 of chlamydial HtrA is critical for substrate binding during both in vitro and in vivo conditions

Journal article

Open access

Peer reviewed

The active site residue V266 of chlamydial HtrA is critical for substrate binding during both in vitro and in vivo conditions

S Gloeckl, J D A Tyndall, S H Stansfield, Peter Timms and W M Huston

Journal of Molecular Microbiology and Biotechnology, Vol.22(1), pp.10-16

2012

DOI: https://doi.org/10.1159/000336312

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Abstract

HtrA

Chlamydia

active-site binding specificity

Bacterial Adenylate Cyclase Two Hybrid (BACTH)

protease

HtrA is a complex, multimeric chaperone and serine protease important for the virulence and survival of many bacteria. Chlamydia trachomatis is an obligate, intracellular bacterial pathogen that is responsible for severe disease pathology. C. trachomatis HtrA (CtHtrA) has been shown to be highly expressed in laboratory models of disease. In this study, molecular modelling of CtHtrA protein active site structure identified putative S1-S3 subsite residues I242, I265, and V266. These residues were altered by site-directed mutagenesis, and these changes were shown to considerably reduce protease activity on known substrates and resulted in a narrower and distinct range of substrates compared to wild type. Bacterial two-hybrid analysis revealed that CtHtrA is able to interact in vivo with a broad range of protein sequences with high affinity. Notably, however, the interaction was significantly altered in 35 out of 69 clones when residue V266 was mutated, indicating that this residue has an important function during substrate binding. © 2012 S. Karger AG, Basel.

Details

Title: The active site residue V266 of chlamydial HtrA is critical for substrate binding during both in vitro and in vivo conditions
Authors: S Gloeckl (Author) - Queensland University of Technology
J D A Tyndall (Author) - University of Otago, New Zealand
S H Stansfield (Author) - Queensland University of Technology
Peter Timms (Author) - Queensland University of Technology
W M Huston (Author) - Queensland University of Technology
Publication details: Journal of Molecular Microbiology and Biotechnology, Vol.22(1), pp.10-16
Publisher: S. Karger AG
Date published: 2012
DOI: 10.1159/000336312
ISSN: 1464-1801
Copyright note: Copyright © 2012 S. Karger AG. This is the Author's Accepted Version of Gloeckl S. Tyndall J.D.A. Stansfield S.H. Timms P. Huston W.M. (2012) The Active Site Residue V266 of Chlamydial HtrA Is Critical for Substrate Binding during both in vitro and in vivo Conditions. J Mol Microbiol Biotechnol 2012;22:10-16 (DOI:10.1159/000336312). The final version is avaialble from: http://dx.doi.org/10.1159/000336312
Organisation Unit: University of the Sunshine Coast, Queensland; Centre for Bioinnovation
Language: English
Record Identifier: 99448847202621
Output Type: Journal article

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