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PDZ domains facilitate binding of high temperature requirement protease A (HtrA) and tail-specific protease (Tsp) to heterologous substrates through recognition of the small stable RNA A (ssrA)-encoded peptide
Journal article   Open access   Peer reviewed

PDZ domains facilitate binding of high temperature requirement protease A (HtrA) and tail-specific protease (Tsp) to heterologous substrates through recognition of the small stable RNA A (ssrA)-encoded peptide

Alison Spiers, Heather K Lamb, Simon Cocklin, Kerry A Wheeler, Jo Budworth, Anna L Dodds, Mark J Pallen, Duncan J Maskell, Ian G Charles and Alastair R Hawkins
The Journal of biological chemistry, v 277(42), pp 39443-39449
18 Oct 2002
DOI: https://doi.org/10.1074/jbc.M202790200
PMID: 12177052
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://doi.org/10.1074/jbc.m202790200View
Published, Version of Record (VoR)CC BY V4.0,  Open
url
https://doi.org/10.1074/jbc.M202790200View
Published, Version of Record (VoR) Open

Abstract

Protein Structure, Tertiary Surface Plasmon Resonance Periplasmic Proteins - chemistry Peptides - chemistry Heat-Shock Proteins - metabolism Glutathione Transferase - metabolism Serine Endopeptidases - chemistry Plasmids - metabolism Recombinant Fusion Proteins - metabolism Protein Transport Salmonella typhimurium - metabolism Chaperonins - metabolism Endopeptidases - chemistry Time Factors Cloning, Molecular Escherichia coli - metabolism Protein Binding Periplasmic Proteins - metabolism Serine Endopeptidases - metabolism Kinetics Binding Sites RNA, Bacterial - metabolism Heat-Shock Proteins - chemistry RNA - metabolism
The Escherichia coli protease HtrA has two PDZ domains, and sequence alignments predict that the E. coli protease Tsp has a single PDZ domain. PDZ domains are composed of short sequences (80-100 amino acids) that have been implicated in a range of protein:protein interactions. The PDZ-like domain of Tsp may be involved in binding to the extreme COOH-terminal sequence of its substrate, whereas the HtrA PDZ domains are involved in subunit assembly and are predicted to be responsible for substrate binding and subsequent translocation into the active site. E. coli has a system of protein quality control surveillance mediated by the ssrA-encoded peptide tagging system. This system tags misfolded proteins or protein fragments with an 11-amino acid peptide that is recognized by a battery of cytoplasmic and periplasmic proteases as a degradation signal. Here we show that both HtrA and Tsp are able to recognize the ssrA-encoded peptide tag with apparent K(D) values of approximately 5 and 390 nm, respectively, and that their PDZ-like domains mediate this recognition. Fusion of the ssrA-encoded peptide tag to the COOH terminus of a heterologous protein (glutathione S-transferase) renders it sensitive to digestion by Tsp but not HtrA. These observations support the prediction that the HtrA PDZ domains facilitate substrate binding and the differential proteolytic responses of HtrA and Tsp to SsrA-tagged glutathione S-transferase are interpreted in terms of the structure of HtrA.

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Web of Science research areas
Biochemistry & Molecular Biology
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