Purification and characterization of a hemolysin produced by Vibrio mimicus

Shin-ichi Miyoshi, Kazuhiro Sasahara, Seiko Akamatsu, M. Monzur Rahman, Takashi Katsu, Ken Ichi Tomochika, Sumio Shinoda

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Vibrio mimicus is a causative agent of human gastroenteritis. This pathogen secretes a pore-forming toxin, V. mimicus hemolysin (VMH), which causes hemolysis by three sequential steps: binding to an erythrocyte membrane, formation of a transmembrane pore, and disruption of the cell membrane. VMH with a molecular mass of 63 kDa was purified by ammonium sulfate precipitation and column chromatography with phenyl Sepharose HP and Superose 6 HR. The hemolytic reaction induced by VMH continued up to disruption of all erythrocytes in the assay system. Moreover, VMH that bound preliminarily to erythrocyte ghosts showed a sufficient ability to attack intact erythrocytes. These results suggest reversible binding of the toxin molecule to the membrane. The final cell-disrupting stage was effectively inhibited by various divalent cations. Additionally, some cations, such as Zn2+ and Cu2+, blocked the pore-forming stage at high concentrations. Although VMH could disrupt all kinds of mammalian erythrocytes tested, those from horses were most sensitive to the hemolysin. Horse erythrocytes were found to have the most toxin-binding sites and to be hemolyzed by the least amount of membrane-bound toxin molecules, suggesting that toxin binding to and pore formation on erythrocytes are more effective in horses than in other mammals. Purified VMH induced fluid accumulation in a ligated rabbit ileal loop in a dose-dependent manner. In addition, the antibody against the hemolysin obviously reduced enteropathogenicity of living V. mimicus cells. These findings clearly demonstrate that VMH is probably involved in the virulence of this human pathogen.

Original languageEnglish
Pages (from-to)1830-1835
Number of pages6
JournalInfection and Immunity
Volume65
Issue number5
Publication statusPublished - 1997

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Erythrocytes
Hemolysin Proteins
Vibrio mimicus
Horses
Erythrocyte Membrane
Membranes
Divalent Cations
Gastroenteritis
Ammonium Sulfate
Hemolysis
Virulence
Vibrio hemolysin
Cations
Chromatography
Mammals
Binding Sites
Cell Membrane
Rabbits
Antibodies

ASJC Scopus subject areas

  • Immunology

Cite this

Miyoshi, S., Sasahara, K., Akamatsu, S., Rahman, M. M., Katsu, T., Tomochika, K. I., & Shinoda, S. (1997). Purification and characterization of a hemolysin produced by Vibrio mimicus. Infection and Immunity, 65(5), 1830-1835.

Purification and characterization of a hemolysin produced by Vibrio mimicus. / Miyoshi, Shin-ichi; Sasahara, Kazuhiro; Akamatsu, Seiko; Rahman, M. Monzur; Katsu, Takashi; Tomochika, Ken Ichi; Shinoda, Sumio.

In: Infection and Immunity, Vol. 65, No. 5, 1997, p. 1830-1835.

Research output: Contribution to journalArticle

Miyoshi, S, Sasahara, K, Akamatsu, S, Rahman, MM, Katsu, T, Tomochika, KI & Shinoda, S 1997, 'Purification and characterization of a hemolysin produced by Vibrio mimicus', Infection and Immunity, vol. 65, no. 5, pp. 1830-1835.
Miyoshi S, Sasahara K, Akamatsu S, Rahman MM, Katsu T, Tomochika KI et al. Purification and characterization of a hemolysin produced by Vibrio mimicus. Infection and Immunity. 1997;65(5):1830-1835.
Miyoshi, Shin-ichi ; Sasahara, Kazuhiro ; Akamatsu, Seiko ; Rahman, M. Monzur ; Katsu, Takashi ; Tomochika, Ken Ichi ; Shinoda, Sumio. / Purification and characterization of a hemolysin produced by Vibrio mimicus. In: Infection and Immunity. 1997 ; Vol. 65, No. 5. pp. 1830-1835.
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