Modulation of Vibrio mimicus hemolysin through limited proteolysis by an endogenous metalloprotease

Tamaki Mizuno, Syed Z. Sultan, Yoshimi Kaneko, Tomonaga Yoshimura, Yoko Maehara, Hiroshi Nakao, Tomofusa Tsuchiya, Sumio Shinoda, Shin-ichi Miyoshi

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Vibrio mimicus is a causative agent of human gastroenteritis and food poisoning, and this species produces an enterotoxic hemolysin (V. mimicus hemolysin) as a virulence determinant. Vibrio mimicus hemolysin is secreted as an 80 kDa precursor, which is later converted to the 66 kDa mature toxin through removal of an N-terminal propeptide via cleavage of the Arg151-Ser152 bond. In this article, we investigate the role of the endogenous metalloprotease (V. mimicus protease) in the maturation of V. mimicus hemolysin. In vitro experiments using purified proteins showed that, although it activated the precursor at the early stage via cleavage of the Asn157-Val158 bond, V. mimicus protease finally converted the activated and physiologically maturated toxin to a 51 kDa protein through removal of the C-terminal polypeptide. This 51 kDa derivative was unable to lyse erythrocytes because of its inability to bind to the erythrocyte membrane. Vibrio mimicus protease-negative strains were found to produce high levels of V. mimicus hemolysin at the logarithmic phase of bacterial growth and maintained high hemolytic activity even at the stationary phase. These findings indicate that, although it is not directly related to toxin maturation in vivo, V. mimicus protease can modulate the activity of V. mimicus hemolysin and/or its precursor through limited proteolysis.

Original languageEnglish
Pages (from-to)825-834
Number of pages10
JournalFEBS Journal
Volume276
Issue number3
DOIs
Publication statusPublished - Feb 2009

Fingerprint

Vibrio mimicus
Proteolysis
Hemolysin Proteins
Metalloproteases
Modulation
Peptide Hydrolases
Foodborne Diseases
Gastroenteritis
Erythrocyte Membrane
Virulence
Proteins
Erythrocytes
Vibrio hemolysin
Derivatives
Membranes
Peptides
Growth

Keywords

  • Hemolysin
  • Metalloprotease
  • Processing
  • Vibrio cholerae
  • Vibrio mimicus

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Modulation of Vibrio mimicus hemolysin through limited proteolysis by an endogenous metalloprotease. / Mizuno, Tamaki; Sultan, Syed Z.; Kaneko, Yoshimi; Yoshimura, Tomonaga; Maehara, Yoko; Nakao, Hiroshi; Tsuchiya, Tomofusa; Shinoda, Sumio; Miyoshi, Shin-ichi.

In: FEBS Journal, Vol. 276, No. 3, 02.2009, p. 825-834.

Research output: Contribution to journalArticle

Mizuno, T, Sultan, SZ, Kaneko, Y, Yoshimura, T, Maehara, Y, Nakao, H, Tsuchiya, T, Shinoda, S & Miyoshi, S 2009, 'Modulation of Vibrio mimicus hemolysin through limited proteolysis by an endogenous metalloprotease', FEBS Journal, vol. 276, no. 3, pp. 825-834. https://doi.org/10.1111/j.1742-4658.2008.06827.x
Mizuno, Tamaki ; Sultan, Syed Z. ; Kaneko, Yoshimi ; Yoshimura, Tomonaga ; Maehara, Yoko ; Nakao, Hiroshi ; Tsuchiya, Tomofusa ; Shinoda, Sumio ; Miyoshi, Shin-ichi. / Modulation of Vibrio mimicus hemolysin through limited proteolysis by an endogenous metalloprotease. In: FEBS Journal. 2009 ; Vol. 276, No. 3. pp. 825-834.
@article{46f36c0e081446828094e7055a155710,
title = "Modulation of Vibrio mimicus hemolysin through limited proteolysis by an endogenous metalloprotease",
abstract = "Vibrio mimicus is a causative agent of human gastroenteritis and food poisoning, and this species produces an enterotoxic hemolysin (V. mimicus hemolysin) as a virulence determinant. Vibrio mimicus hemolysin is secreted as an 80 kDa precursor, which is later converted to the 66 kDa mature toxin through removal of an N-terminal propeptide via cleavage of the Arg151-Ser152 bond. In this article, we investigate the role of the endogenous metalloprotease (V. mimicus protease) in the maturation of V. mimicus hemolysin. In vitro experiments using purified proteins showed that, although it activated the precursor at the early stage via cleavage of the Asn157-Val158 bond, V. mimicus protease finally converted the activated and physiologically maturated toxin to a 51 kDa protein through removal of the C-terminal polypeptide. This 51 kDa derivative was unable to lyse erythrocytes because of its inability to bind to the erythrocyte membrane. Vibrio mimicus protease-negative strains were found to produce high levels of V. mimicus hemolysin at the logarithmic phase of bacterial growth and maintained high hemolytic activity even at the stationary phase. These findings indicate that, although it is not directly related to toxin maturation in vivo, V. mimicus protease can modulate the activity of V. mimicus hemolysin and/or its precursor through limited proteolysis.",
keywords = "Hemolysin, Metalloprotease, Processing, Vibrio cholerae, Vibrio mimicus",
author = "Tamaki Mizuno and Sultan, {Syed Z.} and Yoshimi Kaneko and Tomonaga Yoshimura and Yoko Maehara and Hiroshi Nakao and Tomofusa Tsuchiya and Sumio Shinoda and Shin-ichi Miyoshi",
year = "2009",
month = "2",
doi = "10.1111/j.1742-4658.2008.06827.x",
language = "English",
volume = "276",
pages = "825--834",
journal = "FEBS Journal",
issn = "1742-464X",
publisher = "Wiley-Blackwell",
number = "3",

}

TY - JOUR

T1 - Modulation of Vibrio mimicus hemolysin through limited proteolysis by an endogenous metalloprotease

AU - Mizuno, Tamaki

AU - Sultan, Syed Z.

AU - Kaneko, Yoshimi

AU - Yoshimura, Tomonaga

AU - Maehara, Yoko

AU - Nakao, Hiroshi

AU - Tsuchiya, Tomofusa

AU - Shinoda, Sumio

AU - Miyoshi, Shin-ichi

PY - 2009/2

Y1 - 2009/2

N2 - Vibrio mimicus is a causative agent of human gastroenteritis and food poisoning, and this species produces an enterotoxic hemolysin (V. mimicus hemolysin) as a virulence determinant. Vibrio mimicus hemolysin is secreted as an 80 kDa precursor, which is later converted to the 66 kDa mature toxin through removal of an N-terminal propeptide via cleavage of the Arg151-Ser152 bond. In this article, we investigate the role of the endogenous metalloprotease (V. mimicus protease) in the maturation of V. mimicus hemolysin. In vitro experiments using purified proteins showed that, although it activated the precursor at the early stage via cleavage of the Asn157-Val158 bond, V. mimicus protease finally converted the activated and physiologically maturated toxin to a 51 kDa protein through removal of the C-terminal polypeptide. This 51 kDa derivative was unable to lyse erythrocytes because of its inability to bind to the erythrocyte membrane. Vibrio mimicus protease-negative strains were found to produce high levels of V. mimicus hemolysin at the logarithmic phase of bacterial growth and maintained high hemolytic activity even at the stationary phase. These findings indicate that, although it is not directly related to toxin maturation in vivo, V. mimicus protease can modulate the activity of V. mimicus hemolysin and/or its precursor through limited proteolysis.

AB - Vibrio mimicus is a causative agent of human gastroenteritis and food poisoning, and this species produces an enterotoxic hemolysin (V. mimicus hemolysin) as a virulence determinant. Vibrio mimicus hemolysin is secreted as an 80 kDa precursor, which is later converted to the 66 kDa mature toxin through removal of an N-terminal propeptide via cleavage of the Arg151-Ser152 bond. In this article, we investigate the role of the endogenous metalloprotease (V. mimicus protease) in the maturation of V. mimicus hemolysin. In vitro experiments using purified proteins showed that, although it activated the precursor at the early stage via cleavage of the Asn157-Val158 bond, V. mimicus protease finally converted the activated and physiologically maturated toxin to a 51 kDa protein through removal of the C-terminal polypeptide. This 51 kDa derivative was unable to lyse erythrocytes because of its inability to bind to the erythrocyte membrane. Vibrio mimicus protease-negative strains were found to produce high levels of V. mimicus hemolysin at the logarithmic phase of bacterial growth and maintained high hemolytic activity even at the stationary phase. These findings indicate that, although it is not directly related to toxin maturation in vivo, V. mimicus protease can modulate the activity of V. mimicus hemolysin and/or its precursor through limited proteolysis.

KW - Hemolysin

KW - Metalloprotease

KW - Processing

KW - Vibrio cholerae

KW - Vibrio mimicus

UR - http://www.scopus.com/inward/record.url?scp=58449105159&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=58449105159&partnerID=8YFLogxK

U2 - 10.1111/j.1742-4658.2008.06827.x

DO - 10.1111/j.1742-4658.2008.06827.x

M3 - Article

C2 - 19143841

AN - SCOPUS:58449105159

VL - 276

SP - 825

EP - 834

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 3

ER -