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.
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.
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|Item Type:||Journal Article|
|Keywords:||HtrA, Chlamydia, Active-site binding specificity, Bacterial Adenylate Cyclase, Two Hybrid (BACTH), Protease|
|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|
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