Sequencing, expression and biochemical characterization of the <i>Porphyromonas gingivalis pepO</i> gene encoding a protein homologous to human endothelin‐converting enzyme

Awano, Shuji; Ansai, Toshihiro; Mochizuki, Hajime; Yu, Weixian; Tanzawa, Kazuhiko; Turner, Anthony J.; Takehara, Tadamichi

doi:10.1016/s0014-5793(99)01326-5

Cited by 39 publications

(37 citation statements)

References 35 publications

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“…FTT1209c encodes a predicted M13 metallopeptidase with striking similarity to mammalian endothelin-converting enzyme (ECE-1). FTT1209c has several homologs, including PepO from Porphyromonas gingivalis, which plays a role in host cell invasion (1,4). FTT1209c has been annotated as a pseudogene in the sequenced F. tularensis strain SchuS4 (35).…”

Section: Discussionmentioning

confidence: 99%

Identification of MglA-Regulated Genes Reveals Novel Virulence Factors in Francisella tularensis

et al. 2006

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The facultative intracellular bacterium Francisella tularensis causes the zoonotic disease tularemia. F. tularensis resides within host macrophages in vivo, and this ability is essential for pathogenesis. The transcription factor MglA is required for the expression of several Francisella genes that are necessary for replication in macrophages and for virulence in mice. We hypothesized that the identification of MglAregulated genes in the Francisella genome by transcriptional profiling of wild-type and mglA mutant bacteria would lead to the discovery of new virulence factors utilized by F. tularensis. A total of 102 MglA-regulated genes were identified, the majority of which were positively regulated, including all of the Francisella pathogenicity island (FPI) genes. We mutated novel MglA-regulated genes and tested the mutants for their ability to replicate and induce cytotoxicity in macrophages and to grow in mice. Mutations in MglA-regulated genes within the FPI (pdpB and cds2) as well as outside the FPI (FTT0989, oppB, and FTT1209c) were either attenuated or hypervirulent in macrophages compared to the wild-type strain. All of these mutants exhibited decreased fitness in vivo in competition experiments with wild-type bacteria. We have identified five new Francisella virulence genes, and our results suggest that characterizations of additional MglA-regulated genes will yield further insights into the pathogenesis of this bacterium.

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Section: Discussionmentioning

confidence: 99%

Identification of MglA-Regulated Genes Reveals Novel Virulence Factors in Francisella tularensis

et al. 2006

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“…As determined by a BLAST search of the National Center for Biotechnology Information nonredundant protein sequence database and database of unfinished genomes (1, 2) only a few gram-negative organisms sequenced to date, such as Porphyromonas gingivalis, Shewanella putrefaciens, Caulobacter crescentus, Haemophilus ducreyi, and Legionella pneumoniae, possess regions of DNA with homology to the M13 metallopeptidases. Of these organisms only P. gingivalis has been shown experimentally to produce an enzyme with catalytic activity similar to the mammalian enzymes of this category (4). The majority of bacteria that have sequence homology to the mammalian M13 metallopeptidases are gram-positive and have an intimate association with mammals.…”

Section: Resultsmentioning

confidence: 99%

“…S. parasanguis rPepO was shown here by inhibition studies to be more similar to ECE-1 than to NEP, since it was more sensitive to phosphoramidon than it was to thiorphan. It has been suggested for the same reason that P. gingivalis PepO may also be more akin to ECE-1 than to NEP (4).…”

Section: Resultsmentioning

confidence: 99%

Characterization of a Streptococcal Endopeptidase with Homology to Human Endothelin-Converting Enzyme

2001

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A gene encoding an endopeptidase from Streptococcus parasanguis FW213 has been cloned and shown to have high sequence homology to genes encoding mammalian metalloendopeptidases. The gene, designated S. parasanguis pepO, was cloned into the pET28a expression vector, resulting in a fusion of vector sequences encoding a hexahistidine tag at the carboxyl terminus. The recombinant PepO (rPepO) was expressed in Escherichia coli and purified using an Ni 2؉ affinity column. Polyclonal antiserum to rPepO was raised in rabbits and used to localize FW213 PepO to the cytosol. Southern hybridization and immunoblot analysis revealed that other oral streptococci contain regions of DNA with homology to pepO and produce a protein with antigenic properties similar to that of FW213 PepO. Enzymatic activity assays indicated that only S. parasanguis species possess the ability to cleave metenkephalin, the natural substrate of the human neutral endopeptidase (NEP). Inhibition assays revealed that S. parasanguis PepO is a member of the M13 category of metalloendopeptidases, which includes NEP and endothelin-converting enzyme 1 (ECE-1), an enzyme involved in the maintenance of vascular tone. Thiorphan and phosphoramidon, two specific inhibitors of this category of endopeptidases, were used to determine that S. parasanguis PepO is more similar to ECE-1 than to NEP.

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“…In P. gingivalis, the protease showed activity against Big Endothelin (33), but the results were controversial, and ET-1 generation could not be confirmed by enzyme-linked immunosorbent assay (ELISA). Here, we found that the mycobacterial enzyme Zmp1 does not act as an ECE that converts Big ET-1 into ET-1 mature peptide (Fig.…”

Section: Discussionmentioning

confidence: 99%

The Endothelin System Has a Significant Role in the Pathogenesis and Progression of Mycobacterium tuberculosis Infection

et al. 2014

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d Tuberculosis (TB) remains a major global health problem, and although multiple studies have addressed the relationship between Mycobacterium tuberculosis and the host on an immunological level, few studies have addressed the impact of host physiological responses. Proteases produced by bacteria have been associated with important alterations in the host tissues, and a limited number of these enzymes have been characterized in mycobacterial species. M. tuberculosis produces a protease called Zmp1, which appears to be associated with virulence and has a putative action as an endothelin-converting enzyme. Endothelins are a family of vasoactive peptides, of which 3 distinct isoforms exist, and endothelin 1 (ET-1) is the most abundant and the bestcharacterized isoform. The aim of this work was to characterize the Zmp1 protease and evaluate its role in pathogenicity. Here, we have shown that M. tuberculosis produces and secretes an enzyme with ET-1 cleavage activity. These data demonstrate a possible role of Zmp1 for mycobacterium-host interactions and highlights its potential as a drug target. Moreover, the results suggest that endothelin pathways have a role in the pathogenesis of M. tuberculosis infections, and ETA or ETB receptor signaling can modulate the host response to the infection. We hypothesize that a balance between Zmp1 control of ET-1 levels and ETA/ ETB signaling can allow M. tuberculosis adaptation and survival in the lung tissues.

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Sequencing, expression and biochemical characterization of the Porphyromonas gingivalis pepO gene encoding a protein homologous to human endothelin‐converting enzyme

Cited by 39 publications

References 35 publications

Identification of MglA-Regulated Genes Reveals Novel Virulence Factors in Francisella tularensis

Identification of MglA-Regulated Genes Reveals Novel Virulence Factors in Francisella tularensis

Characterization of a Streptococcal Endopeptidase with Homology to Human Endothelin-Converting Enzyme

The Endothelin System Has a Significant Role in the Pathogenesis and Progression of Mycobacterium tuberculosis Infection

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