The protease from Vibrio Cholerae nicks arginine at position 192 from the N-terminus of the heat-labile enterotoxin a subunit from enterotoxigenic Escherichia Coli

Ichinose, Yuki; Tsuji, Takao; Ehara, Masahiko; Miyama, Akio; Naito, Toshiki

doi:10.1007/bf00145394

Cited by 11 publications

(8 citation statements)

References 20 publications

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“…One is that the level of production of the nicking enzyme by this strain might be lower than those of other strains, as some proteases were reported to be involved in the nicking of the CT A subunit. The extent of protein digestion of the culture supernatant was almost identical to those of other strains, as measured by the skim-milk method (14); hence, other nicking enzymes may be involved in nicking in this strain. The second explanation for strain No.…”

supporting

confidence: 63%

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A classical strain of Vibrio cholerae with diminished ability to process the proteolytically sensitive site in the A subunit of cholera toxin

et al. 1996

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Vibrio cholerae O1, No. 31, a strain isolated from a patient with mild diarrhea, produced mainly the unnicked cholera toxin. The amount of toxin that had accumulated in the cells was approximately 200 times lower than that secreted into the culture medium. When the unnicked toxin was purified by three successive column chromatographies and then extracted from the polyacrylamide gel, the unnicked toxin showed two bands corresponding to the A and B subunits by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the A1 fragment was detected by trypsinization. Biological and enzymatic activities of the purified toxin with trypsinization were identical to those of cholera toxin from V. cholerae 569B as seen in the rabbit skin permeability test and the NAD:agmatine ADP-ribosyltransferase assay. DNA sequences of the A and B subunits were identical to those of the A-and B-subunit genes from the El Tor 2125 and classical 0395 strains, respectively. These data suggest that the wild V. cholerae strain, No. 31, produces a toxin identical to toxins previously reported in the literature and secretes it without accumulation in the cell, as is the case with other strains. However, strain No. 31's ability to nick the toxin is diminished compared with such abilities of other strains.

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supporting

confidence: 63%

“…An endogenous enzyme like hemagglutinin/protease from V. cholerae might be involved in this cleavage of the A subunit (7,14). However, it has not been determined whether these enzymes are essential for the nicking of the toxin or where the toxin is nicked.…”

mentioning

confidence: 99%

A classical strain of Vibrio cholerae with diminished ability to process the proteolytically sensitive site in the A subunit of cholera toxin

et al. 1996

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“…Pseudomonas aeruginosa elastase (33 kDa) (15) and alkaline proteinase (48 kDa) (16) were obtained from Nagase Biochemicals, Osaka, Japan. Vibrio cholerae HA/proteinase (32 kDa) was purified according to a previously given method (17). Serratia marcescens 56-kDa metalloproteinase and 73-kDa thiol proteinase were purified as described previously (18).…”

Section: Methodsmentioning

confidence: 99%

Activation of Human Matrix Metalloproteinases by Various Bacterial Proteinases

Okamoto¹,

Akaike²,

Suga³

et al. 1997

Journal of Biological Chemistry

144

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Matrix metalloproteinases (MMPs) are zinc-containing proteinases that participate in tissue remodeling under physiological and pathological conditions. To test the involvement of bacterial proteinases in tissue injury during bacterial infections, we investigated the activation potential of various bacterial proteinases against precursors of MMPs (proMMPs) purified from human neutrophils (proMMP-8 and -9) and from human fibrosarcoma cells (proMMP-1). Each proMMP was subjected to treatment with a series of bacterial proteinases at molar ratios of 0.01-0.1 (bacterial proteinase to proMMP), and activities of MMPs generated were determined. Among six different bacterial proteinases, thermolysin family enzymes (family M4) such as Pseudomonas aeruginosa elastase, Vibrio cholerae proteinase, and thermolysin strongly activated all three proMMPs via limited proteolysis to generate active forms of the MMPs. N-terminal sequence analysis of the active MMPs revealed that cleavage occurred at the Val 82 -Leu 83 and Thr 90 -Phe 91 bonds of proMMP-1 and proMMP-9, respectively, which are located near the N terminus of the catalytic domain of MMPs. In contrast, Serratia 56-kDa proteinase and Pseudomonas alkaline proteinase, both of which are classified as members of the serralysin subfamily of zinc metalloproteinases (family M10), and Serratia 73-kDa thiol proteinase did not evidence proteolytic processing or activation of proMMP-1, -8, and -9 under these experimental conditions. These results indicate that bacterial proteinases may play an important role in tissue destruction and disintegration of extracellular matrix at the site of infections.Collagen, one of the major structural components of the extracellular matrix, has a triple-helical structure and exhibits resistance to proteolytic cleavage by endogenous and exogenous proteinases (1) except for matrix metalloproteinases (MMPs) 1 such as human neutrophil collagenase (MMP-8). Bacterial proteinases have been suggested to mediate direct tissue destruction, resulting in impairment of host defense mechanisms in septic foci (2-4). Most bacterial proteinases, however, have weak degradative activity against collagen (1, 5). Thus, the mechanism of extracellular matrix destruction at the site of bacterial infection is poorly understood.MMPs, a family of zinc neutral endopeptidases, are secreted by a variety of cells as inactive precursors (proMMPs) and degrade a series of collagens (6). Two distinct proMMPs (proMMP-8 and neutrophil progelatinase, proMMP-9) are synthesized and secreted extracellularly from specific granules of human neutrophils after membrane stimulation (7, 8). Macrophages and fibroblasts produce interstitial procollagenase (proMMP-1) (6, 9) and 92-kDa progelatinase (proMMP-9), whose expressions vary constitutively or inducibly after stimulation with proinflammatory cytokines and lipopolysaccharide (10 -12). MMP-8 and -1 specifically cleave native triplehelical type I collagen into two major fragments, one-fourth and three-fourths the size of native collagen, respect...

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“…The 20-60% ammonium sulfate-insoluble materials were suspended in 25 mM PB and dialyzed against the same buffer. Dialysates were centrifuged at 100000 xg for 1 h and the supernatant was applied to a Biogel A5m column and a TSK gel G-3000 SW column on HPLC as described previously [9]. Fractions with protease activity, which showed a single peak with HPLC, were used as purified protease.…”

Section: Purification Of Protease From Vibrio Cholerae 01mentioning

confidence: 99%

The effect on enterotoxicity of protease purified from Vibrio cholerae O1

Ichinose

1994

FEMS Microbiology Letters

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The effect on enterotoxicity of protease purified from Vibrio cholerae O1 was investigated by the inoculation of live vibrio cells into protease-treated loops of the ileal loop model. Fluid accumulation ratios in the protease-treated loops were elevated in a dose-dependent manner by challenge with live vibrio cells but not by that with toxin. An enhancement effect of protease on enterotoxicity was observed in both serotypes of V. cholerae O1 and V. cholerae non-O1. It is suggested, therefore, that the enterotoxicity was enhanced by treatment with protease when live vibrio cells were inoculated into the ileal loops of rabbits.

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The protease from Vibrio Cholerae nicks arginine at position 192 from the N-terminus of the heat-labile enterotoxin a subunit from enterotoxigenic Escherichia Coli

Cited by 11 publications

References 20 publications

A classical strain of Vibrio cholerae with diminished ability to process the proteolytically sensitive site in the A subunit of cholera toxin

A classical strain of Vibrio cholerae with diminished ability to process the proteolytically sensitive site in the A subunit of cholera toxin

Activation of Human Matrix Metalloproteinases by Various Bacterial Proteinases

The effect on enterotoxicity of protease purified from Vibrio cholerae O1

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