PDZ domains determine the native oligomeric structure of the DegP (HtrA) protease

Sassoon, Nathalie; Arié, Jean‐Philippe; Betton, Jean‐Michel

doi:10.1046/j.1365-2958.1999.01505.x

Cited by 43 publications

(42 citation statements)

References 27 publications

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“…The DegP protease has two so-called PDZ domains (postsynaptic density 95, discs large, ZO-1) located downstream of the catalytic serine residue (33). PDZ domains have been implicated in both substrate recognition and protein multimerization in a number of proteins (26,40,46). The carboxylterminal motif of pilin subunits has some homology to the motif reportedly recognized by PDZ domains (26,46).…”

Section: Resultsmentioning

confidence: 99%

“…3 and 5). Whether the noncleavable peptide (SPCJ-1) stimulates multimerization of DegP, the multimer is the proposed active state of the enzyme (22,33,40), or it represents a signal that unfolded substrate is present and activates DegP through another mechanism is unclear. As described in detail in two recent papers (5,41) and as supported by earlier genetic data (3,14,24,44,47,55), the carboxyl terminus of pilin subunits is an integral component of the recognition site for chaperone-subunit complex formation.…”

Section: Discussionmentioning

confidence: 99%

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Escherichia coli DegP Protease Cleaves between Paired Hydrophobic Residues in a Natural Substrate: the PapA Pilin

et al. 2002

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The DegP protein, a multifunctional chaperone and protease, is essential for clearance of denatured or aggregated proteins from the inner-membrane and periplasmic space in Escherichia coli. To date, four natural targets for DegP have been described: colicin A lysis protein, pilin subunits and MalS from E. coli, and high-molecular-weight adherence proteins from Haemophilus influenzae. In vitro, DegP has shown weak protease activity with casein and several other nonnative substrates. We report here the identification of the major pilin subunit of the Pap pilus, PapA, as a natural DegP substrate and demonstrate binding and proteolysis of this substrate in vitro. Using overlapping peptide arrays, we identified three regions in PapA that are preferentially cleaved by DegP. A 7-mer peptide was found to be a suitable substrate for cleavage by DegP in vitro. In vitro proteolysis of model peptide substrates revealed that cleavage is dependent upon the presence of paired hydrophobic amino acids; moreover, cleavage was found to occur between the hydrophobic residues. Finally, we demonstrate that the conserved carboxyl-terminal sequence in pilin subunits, although not a cleavage substrate for DegP, activates the protease and we propose that the activating peptide is recognized by DegP's PDZ domains.The DegP (HtrA, protease Do) protease is a multifunctional protein essential for the removal of misfolded and aggregated proteins in the periplasm of Escherichia coli. DegP is one of several dozen proteases in E. coli and is known to have homologues in virtually all gram-negative bacteria, cyanobacteria, and mycobacteria as well as in higher-order organisms such as yeasts and humans (33). DegP homologues have recently been described for a number of gram-positive bacteria (18,32,36,45). There are also two homologues of DegP, referred to as DegQ and DegS, in E. coli (22,54). DegP has been rediscovered several times, as is revealed by the nomenclature (30,33,42). The DegP (for "degradation") nomenclature refers to the initial mapping of a mutation in E. coli that allowed the accumulation of unstable fusion proteins in the periplasm (50, 51). The HtrA (for "heat shock regulated") designation indicates that a transposon insertion in the same gene resulted in a temperature-sensitive growth phenotype (28). Lastly, DegP was also designated protease Do, again as a mutation that conferred a temperature-sensitive growth phenotype in E. coli (42). The DegP designation is used throughout this report.Purified DegP exhibited functional protease activity in in vitro assays using casein as a substrate, although its activity on this substrate was weak (29). Lipinska et al. (29) demonstrated that the activity with casein was inhibitable by diisopropyl fluorophosphate and not by any other known protease inhibitors, suggesting that DegP contains an active site serine residue. Interestingly, DegP is not inhibited by the classic serine protease inhibitor, phenylmethyl-sulfonyl fluoride, suggesting differences in the mode of action of DegP (21, 29). Si...

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Escherichia coli DegP Protease Cleaves between Paired Hydrophobic Residues in a Natural Substrate: the PapA Pilin

et al. 2002

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“…The HtrA family shares a highly conserved trypsinlike serine proteinase domain and one or two C-terminal PDZ domains (28). The chaperone and serine proteinase HtrA is a virulence factor in diverse pathogens (29), but its potential substrates and role in H. pylori remain to be elucidated.…”

Section: Trypsin/ Par Pathway In Reflux Esophagitismentioning

confidence: 99%

Basic and Translational Research on Proteinase-Activated Receptors: Implication of Proteinase/Proteinase-Activated Receptor in Gastrointestinal Inflammation

Yoshida

Yoshikawa

2008

J Pharmacol Sci

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Abstract. Recently, the role of serine proteinases in the pathogenesis of inflammation and autoimmune diseases via interaction with the proteinase-activated receptor (PAR) has attracted attention. Activation of PAR has a pro-inflammatory effect through the overproduction of inflammatory cytokines such as interleukin (IL)-6 and IL-8. PAR 2 activation in human esophageal epithelial cells by trypsin induces NFκB-and AP-1-dependent IL-8 production in association with activation of p38 MAPK and ERK1 / 2, suggesting that esophageal inflammation may be induced by PAR 2 activation via reflux of trypsin. It has been also proposed that Helicobacter pylori (H. pylori) induces PAR expression in the gastric epithelial cells and H. pylori-derived serine proteinase promotes IL-8 production via PAR in the epithelial cells. In addition, an increase of PAR-dependent IL-8 production has been observed in H. pylori-infected human gastric mucosa, suggesting an important role for PAR 2 in the modulation of gastric inflammation associated with H. pylori. Recent studies have strongly indicated that tryptase and PAR are implicated in the pathogenesis of inflammatory bowel disease and experimental colitis. We demonstrated that anti-tryptase therapy may become a new therapeutic strategy in human ulcerative colitis. Thus, the role of PAR in the gastrointestinal tract has been gradually clarified, but further investigations are needed because the receptor has a variety of functions.

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“…It has an oligometric structure that appears to be essential for the proteolytic activity (Kim et al, 1999;Sassoon et al, 1999). DegP eliminates a variety of aberrant proteins arising in the periplasmic compartment (Pallen and Wren, 1997).…”

Section: Introductionmentioning

confidence: 99%

Efficient export of alkaline phosphatase overexpressed from a multicopy plasmid requires degP, a gene encoding a periplasmic protease of Escherichia coli.

Kadokura

Kawasaki

Yoda

et al. 2001

J. Gen. Appl. Microbiol.

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Escherichia coliDegP is an inducible serine protease which is involved in the breakdown of abberant proteins arising in the periplasmic compartment. Overexpression of alkaline phosphatase (PhoA) increased transcription of degP by twofold. To examine the significance of its induction, we overexpressed PhoA in a mutant strain deficient in the degP gene. Upon PhoA overexpression, the degP mutant produced a smaller amount of active PhoA, about one half of the enzymatic activity of its isogenic wild-type strain, and accumulated a larger amount of its precursor, indicating that degP is required for efficient export of overexpressed PhoA. Pulse-chase experiment showed that PhoA overexpression in the absence of degP causes a severe defect in the export of several proteins tested. Examination of the synthesis and the accumulation of the phoA gene products revealed that a part of them, synthesized in the wild-type strain, undergoes relatively rapid proteolysis and that degP is necessary for such a process. From these results, we discuss a possible role of DegP in facilitating protein export under stress conditions.

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PDZ domains determine the native oligomeric structure of the DegP (HtrA) protease

Cited by 43 publications

References 27 publications

Escherichia coli DegP Protease Cleaves between Paired Hydrophobic Residues in a Natural Substrate: the PapA Pilin

Escherichia coli DegP Protease Cleaves between Paired Hydrophobic Residues in a Natural Substrate: the PapA Pilin

Basic and Translational Research on Proteinase-Activated Receptors: Implication of Proteinase/Proteinase-Activated Receptor in Gastrointestinal Inflammation

Efficient export of alkaline phosphatase overexpressed from a multicopy plasmid requires degP, a gene encoding a periplasmic protease of Escherichia coli.

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