The FAXWXXT motif in the carboxyl terminus of <i>Vibrio mimicus</i> metalloprotease is involved in binding to collagen

Lee, Jonghee; Ahn, Seungjoon; Lee, Eun-Mi; Jeong, Seung-Ha; Kim, Young‐Ok; Lee, Sang-Jun; Kong, In‐Soo

doi:10.1016/j.febslet.2005.03.062

Cited by 24 publications

(31 citation statements)

References 30 publications

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“…In addition, two FAXWXXT motifs, FDQWAQS and FSAWLDT (conserved residues are in boldface), which are implicated in collagen binding (32), and two bacterial prepeptidase C-terminal (PPC) domains also were identified at amino acids 537 to 543, 555 to 561, 612 to 686, and 731 to 804, respectively. A comparison of the amino acid sequences of class II metalloproteases in different Vibrio species showed that VC1650 displays the highest similarity (over 70%) to the 61-kDa collagenase VMC from V. mimicus and 63-kDa PrtVp protease (VPPRT) from V. parahaemolyticus 93 (31)(32)(33)(34).…”

Section: Resultsmentioning

confidence: 99%

“…Very limited information is available regarding the tertiary structure of the Vibrio collagenases belonging to the M9A subfamily, and the function and biochemical properties of their domains, as well as the physiological importance of the C-terminal processing, remain to be addressed. It can be speculated that the removal of both PPC domains exposes the repeated FAXWXXT motif, which has been implicated in binding to insoluble type I collagen (32), and provides a better interaction with the substrate.…”

Section: Discussionmentioning

confidence: 99%

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A Metalloprotease Secreted by the Type II Secretion System Links Vibrio cholerae with Collagen

et al. 2015

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bVibrio cholerae is autochthonous to various aquatic niches and is the etiological agent of the life-threatening diarrheal disease cholera. The persistence of V. cholerae in natural habitats is a crucial factor in the epidemiology of cholera. In contrast to the well-studied V. cholerae-chitin connection, scarce information is available about the factors employed by the bacteria for the interaction with collagens. Collagens might serve as biologically relevant substrates, because they are the most abundant protein constituents of metazoan tissues and V. cholerae has been identified in association with invertebrate and vertebrate marine animals, as well as in a benthic zone of the ocean where organic matter, including collagens, accumulates. Here, we describe the characterization of the V. cholerae putative collagenase, VchC, encoded by open reading frame VC1650 and belonging to the subfamily M9A peptidases. Our studies demonstrate that VchC is an extracellular collagenase degrading native type I collagen of fish and mammalian origin. Alteration of the predicted catalytic residues coordinating zinc ions completely abolished the protein enzymatic activity but did not affect the translocation of the protease by the type II secretion pathway into the extracellular milieu. We also show that the protease undergoes a maturation process with the aid of a secreted factor(s). Finally, we propose that V. cholerae is a collagenovorous bacterium, as it is able to utilize collagen as a sole nutrient source. This study initiates new lines of investigations aiming to uncover the structural and functional components of the V. cholerae collagen utilization program. Members of the genus Vibrio are important inhabitants of the marine coastal waters, estuaries, and ocean sediments, with some pathogenic species also causing wound infections, primary septicemia, gastroenteritis, and diarrhea in humans (1, 2). Among the more than 200 serogroups of V. cholerae, only O1 and O139 are recognized as agents of cholera, a potentially life-threatening diarrheal disease. Cholera remains a significant public health problem by affecting millions of people annually, predominantly in developing countries that have limited clean water supplies and poor sanitation (3). Infection of the human host occurs upon ingestion of water or food contaminated with the bacterium. Following colonization of the small intestine, V. cholerae utilizes a multicomponent secretory pathway, the type II secretion system (T2SS), to release cholera toxin. Cholera toxin induces acute diarrhea in people infected with cholera, enabling the bacteria to escape from the host into the aquatic reservoir (4). During interepidemic periods, V. cholerae persists freely or in association with various aquatic organisms, including copepods, insect egg masses, shellfish, and vertebrate fish (5-7). Extracellular proteins, including those secreted by the T2SS, have been implicated in facilitating the fitness of V. cholerae in both the human host and aquatic niches (4,6,8). The array of extrace...

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A Metalloprotease Secreted by the Type II Secretion System Links Vibrio cholerae with Collagen

et al. 2015

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“…However, the substratebinding domains of bacterial MMPs are much smaller than the substrate-binding domains of mammalian MMPs. It has been demonstrated that ϳ33 amino acids of the Vibrio mimicus metalloprotease C-terminal domain or ϳ100 amino acids of the Clostridium histolyticum metalloprotease C-terminal domain are necessary for substrate binding, compared to ϳ200 amino acid residues for the hemopexin-like domain in mammalian MMPs (17,36). Thus, we hypothesize that the C-terminal amino acid residues are responsible for binding to the BFT substrate.…”

Section: Vol 74 2006 Structure-function Properties Of B Fragilis Tmentioning

confidence: 99%

“…Mammalian MMPs are larger than bacterial MMPs and contain various arrangements of substrate-binding domains, including fibronectin-like and hemopexin domains, in their C-terminal regions (26). Recently, studies have shown that, similar to the C terminus of mammalian MMPs, the C-terminal regions of some bacterial MMPs contain a collagen-binding domain, as shown by loss of collagenase activity after deletion of the C-terminal region (17,18,34). However, the substratebinding domains of bacterial MMPs are much smaller than the substrate-binding domains of mammalian MMPs.…”

Section: Vol 74 2006 Structure-function Properties Of B Fragilis Tmentioning

confidence: 99%

“…In recent studies, we have demonstrated that a series of single point mutations in the zinc-binding metalloprotease motif do not affect BFT processing but do reduce or eliminate BFT biologic activity in vitro (5). Recently, studies have also shown that the C-terminal regions of some bacterial MMPs are necessary for substrate binding, as shown by loss of activity after deletion of the Cterminal region (17,18,34). In this study, we evaluated the role of the C-terminal region in BFT activity, processing, and secretion.…”

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The C-Terminal Region of Bacteroides fragilis Toxin Is Essential to Its Biological Activity

et al. 2006

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To evaluate the role of the C-terminal region in Bacteroides fragilis toxin (BFT) activity, processing, and secretion, sequential C-terminal truncation and point mutations were created by site-directed mutagenesis. Determination of BFT activity on HT29/C1 cells, cleavage of E-cadherin, and the capacity to induce interleukin-8 secretion by wild-type BFT and C-terminal deletion mutants showed that deletion of only 2 amino acid residues at the C terminus significantly reduced BFT biological activity and deletion of eight or more amino acid residues obliterated BFT biologic activity. Western blot and reverse transcription-PCR analyses indicated that BFT mutants lacking seven or fewer amino acid residues in the C-terminal region are processed and expressed similar to wild-type BFT. However, BFT mutants lacking eight or more amino acids at the C terminus are expressed similar to wild-type BFT but are unstable. We concluded that the C terminus of BFT is not tolerant of modest amino acid deletions, suggesting that it is biologically important for BFT activity.Enterotoxigenic Bacteroides fragilis (ETBF) is strongly linked epidemiologically to diarrheal disease in livestock, young children, and adults (22,23,27,28,30,35,40). The only recognized virulence factor of ETBF is a secreted 20-kDa zinc-dependent metalloprotease termed B. fragilis toxin (BFT) (19). BFT causes fluid accumulation in ligated intestinal loops of lambs, rats, rabbits, and calves (23,24,31). In vitro, BFT alters the morphology of certain human intestinal carcinoma cell lines, particularly cell line HT29/C1 (3,15,31,35). Subconfluent HT29/C1 cells treated with BFT develop striking changes in morphology, including loss of cell-to-cell attachments, rounding, swelling, and, in some cases, pyknosis. The mechanism of action and morphological changes stimulated by BFT are mediated, in part, by cleavage of the zonula adherens protein, E-cadherin (38). Recently, ETBF strains have also been associated with active inflammatory bowel disease and colorectal cancer (1,25,33). We and other workers (13,29,39) have shown that BFT stimulates interleukin-8 (IL-8) secretion by intestinal cells (HT29, T84, and Caco-2 cells) in vitro.Three highly related isotypes of BFT have been identified (termed BFT-1, BFT-2, and BFT-3) (4,8,12,37). All BFTs appear to be structurally similar. BFT is synthesized as a 44-kDa precursor (397 amino acid residues) containing the following three consecutive peptide domains: (i) a presignal sequence (18 amino acid residues), (ii) a propeptide (193 amino acid residues), and (iii) a mature protein (186 amino acid residues) (8, 14). The 44-kDa precursor protein is processed to a 20-kDa mature BFT that is secreted into the culture supernatant.Based on sequence analysis, BFT is predicted to be a member of the metzincin superfamily of zinc-dependent metalloprotease enzymes (19). Members of this superfamily contain an elongated zinc-binding metalloprotease motif (HEXXHX XGXXH) and present a perfectly superimposable methionine residue close to the zinc-b...

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Regulation of Vibrio mimicus metalloprotease (VMP) production by the quorum‐sensing master regulatory protein, LuxR

Abdel-Sattar

Miyoshi

Elgaml

2016

J. Basic Microbiol.

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Vibrio mimicus is an estuarine bacterium, while it can cause severe diarrhea, wound infection, and otitis media in humans. This pathogen secretes a relatively important toxin named V. mimicus metalloprotease (VMP). In this study, we clarified regulation of the VMP production according to the quorum-sensing master regulatory protein named LuxR. First, the full length of luxR gene, encoding LuxR, was detected in V. mimicus strain E-37, an environmental isolate. Next, the putative consensus binding sequence of LuxR protein could be detected in the upstream (promoter) region of VMP encoding gene, vmp. Finally, the effect of disruption of luxR gene on the expression of vmp and production of VMP was evaluated. Namely, the expression of vmp was significantly diminished by luxR disruption and the production of VMP was severely altered. Taken together, here we report that VMP production is under the positive regulation of the quorum-sensing master regulatory protein, LuxR.

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The FAXWXXT motif in the carboxyl terminus of Vibrio mimicus metalloprotease is involved in binding to collagen

Cited by 24 publications

References 30 publications

A Metalloprotease Secreted by the Type II Secretion System Links Vibrio cholerae with Collagen

A Metalloprotease Secreted by the Type II Secretion System Links Vibrio cholerae with Collagen

The C-Terminal Region of Bacteroides fragilis Toxin Is Essential to Its Biological Activity

Regulation of Vibrio mimicus metalloprotease (VMP) production by the quorum‐sensing master regulatory protein, LuxR

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