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

Gloeckl, Sarina, Tyndall, Joel D. A., Stansfield, Scott H., Timms, Peter, & Huston, Wilhelmina M. (2012) The active site residue V266 of Chlamydial HtrA is critical for substrate binding during both in vitro and in vivo conditions. Journal of Molecular Microbiology and Biotechnology, 22(1), pp. 10-16.

View at publisher

Abstract

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.

Impact and interest:

3 citations in Scopus
Search Google Scholar™
4 citations in Web of Science®

Citation counts are sourced monthly from Scopus and Web of Science® citation databases.

These databases contain citations from different subsets of available publications and different time periods and thus the citation count from each is usually different. Some works are not in either database and no count is displayed. Scopus includes citations from articles published in 1996 onwards, and Web of Science® generally from 1980 onwards.

Citations counts from the Google Scholar™ indexing service can be viewed at the linked Google Scholar™ search.

Full-text downloads:

105 since deposited on 12 Dec 2012
16 in the past twelve months

Full-text downloads displays the total number of times this work’s files (e.g., a PDF) have been downloaded from QUT ePrints as well as the number of downloads in the previous 365 days. The count includes downloads for all files if a work has more than one.

ID Code: 54542
Item Type: Journal Article
Refereed: Yes
Keywords: HtrA, Chlamydia, Active-site binding specificity, Bacterial Adenylate Cyclase, Two Hybrid (BACTH), Protease
DOI: 10.1159/000336312
ISSN: 1660-2412
Subjects: Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000)
Australian and New Zealand Standard Research Classification > TECHNOLOGY (100000)
Divisions: Current > Schools > School of Biomedical Sciences
Current > QUT Faculties and Divisions > Faculty of Health
Current > Institutes > Institute of Health and Biomedical Innovation
Copyright Owner: Copyright 2012 S. Karger AG, Basel
Deposited On: 12 Dec 2012 03:20
Last Modified: 03 Jun 2016 02:24

Export: EndNote | Dublin Core | BibTeX

Repository Staff Only: item control page