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.
authors.affiliation: Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Japan.
authors.forename: Tamaki
authors.initials: T
authors.lastname: Mizuno
authors.forename: Syed Z
authors.initials: SZ
authors.lastname: Sultan
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Kaneko
authors.forename: Tomonaga
authors.initials: T
authors.lastname: Yoshimura
authors.forename: Yoko
authors.initials: Y
authors.lastname: Maehara
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nakao
authors.forename: Tomofusa
authors.initials: T
authors.lastname: Tsuchiya
authors.forename: Sumio
authors.initials: S
authors.lastname: Shinoda
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Miyoshi
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D045726:Metalloproteases / Q000378:metabolism*; D008970:Molecular Weight; D011485:Protein Binding; D044164:Vibrio mimicus / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D006460:Hemolysin Proteins; C073845:hemolysin, Vibrio; D045726:Metalloproteases