ID | 64008 |
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Miyoshi, Shin-ichi
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
Toko, Norie
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
Dodo, Tetsuya
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
Nanko, Ayako
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
Mizuno, Tamaki
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
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Abstract | Vibrio mimicus is a bacterium that causes gastroenteritis in humans. This pathogen produces an enterotoxic hemolysin called V. mimicus hemolysin (VMH), which is secreted extracellularly as an inactive 80-kDa protoxin and converted to a 66-kDa mature toxin through cleavage between Arg(151) and Ser(152). The 56-kDa serine protease termed V. mimicus trypsin-like protease (VmtA) is known to mediate this maturating process. However, some strains including strain ES-20 does not possess the vmtA gene. In the present study, the vmtA-negative strains were found to have a replaced gene that encodes a 43-kDa (403 aa) precursor of a serine protease designated by VmtX (V. mimicus trypsin-like protease X). To examine whether VmtX is also involved in the maturation of VMH, VmtX was isolated from the culture supernatant of V. mimicus strain NRE-20, a metalloprotease-negative mutant constructed from strain ES-20. Concretely, the culture supernatant was fractionated with 70% saturated ammonium sulfate and subjected to affinity column chromatography using a HiTrap Benzamidine FF column. The analysis of the N-terminal amino acid sequences of the proteins in the obtained VmtX preparation indicated that the 39-kDa protein was active VmtX consisting of 371 aa (Ile(33)-Ser(403)). The VmtX preparation was found to activate pro-VMH through generation of the 66-kDa protein. Additionally, treatment of the VmtX preparation with serine protease inhibitors, such as leupeptin and phenylmethylsulfonyl fluoride, significantly suppressed the activities to hydrolyze the specific peptide substrate and to synthesize the 66-kDa toxin. These findings indicate that VmtX is the second protease that mediats the maturation of VMH.
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Keywords | Vibrio mimicus
Serine protease
Hemolysin
Maturation
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Note | This version of the article has been accepted for publication, after peer review (when applicable) and is subject to Springer Nature’s AM terms of use, but is not the Version of Record and does not reflect post-acceptance improvements, or any corrections. The Version of Record is available online at: http://dx.doi.org/10.1007/s11274-022-03436-9
This fulltext is available in Oct. 2023.
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Published Date | 2022-10-22
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Publication Title |
World Journal of Microbiology and Biotechnology
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Volume | volume38
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Issue | issue12
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Publisher | Springer Science and Business Media LLC
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Start Page | 241
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ISSN | 0959-3993
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NCID | AA10811107
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Content Type |
Journal Article
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language |
English
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OAI-PMH Set |
岡山大学
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Copyright Holders | © The Author(s), under exclusive licence to Springer Nature B.V. 2022
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File Version | author
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Related Url | isVersionOf https://doi.org/10.1007/s11274-022-03436-9
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Citation | Miyoshi, Si., Toko, N., Dodo, T. et al. Second extracellular protease mediating maturation of Vibrio mimicus hemolysin. World J Microbiol Biotechnol 38, 241 (2022). https://doi.org/10.1007/s11274-022-03436-9
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