title: Mode of primary binding to target membranes and pore formation induced by Vibrio cholerae cytolysin (hemolysin).
abstract: Vibrio cholerae cytolysin (VCC) is produced by many non-choleratoxigenic strains of V. cholerae, and possibly represents a relevant pathogenicity determinant of these bacteria. The protein is secreted as a pro-toxin that is proteolytically cleaved to yield the active toxin with a molecular mass of approximately 63 kDa. We here describe a simple procedure for preparative isolation of mature VCC from bacterial culture supernatants, and present information on its mode of binding and pore formation in biological membranes. At low concentrations, toxin monomers interact with a high-affinity binding site on highly susceptible rabbit erythrocytes. This as yet unidentified binding site is absent on human erythrocytes, which are less susceptible to the toxin action. At higher concentrations, binding of the toxin occurs to both rabbit and human erythrocytes in a non-saturable manner. Cell-bound toxin monomers oligomerize to form supramolecular structures that are seen in the electron microscope as apparently hollow funnels, and oligomerization correlates functionally with the appearance of small transmembrane pores. Osmotic protection experiments indicate that the toxin channels are of finite size with a diameter of 1-2 nm. The mode of action of VCC closely resembles that of classical pore-forming toxins such as staphylococcal alpha-toxin and the aerolysin of Aeromonas hydrophila.
authors.affiliation: Institute of Medical Microbiology and Hygiene, University of Mainz, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Zitzer
authors.forename: M
authors.initials: M
authors.lastname: Palmer
authors.forename: U
authors.initials: U
authors.lastname: Weller
authors.forename: T
authors.initials: T
authors.lastname: Wassenaar
authors.forename: C
authors.initials: C
authors.lastname: Biermann
authors.forename: J
authors.initials: J
authors.lastname: Tranum-Jensen
authors.forename: S
authors.initials: S
authors.lastname: Bhakdi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003603:Cytotoxins / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011817:Rabbits; D014734:Vibrio cholerae / Q000472:pathogenicity*
chemical_list: D003603:Cytotoxins