title: Lumen geometry of ion channels formed by Vibrio cholerae EL Tor cytolysin elucidated by nonelectrolyte exclusion.
abstract: Vibrio cholerae EL Tor cytolysin, a water-soluble protein with a molecular mass of 63 kDa, forms small pores in target cell membranes. In this communication, planar lipid bilayers under voltage clamp conditions were used to investigate the geometric properties of the pores. It was established that all cytolysin channels were inserted into membranes with the same orientation. Sharp asymmetry in the I-V curve of fully open cytolysin channels persisting at high electrolyte concentrations indicated asymmetry in the geometry of the channel lumen. Using the nonelectrolyte exclusion method, evidence was obtained that the cis opening of the channel had a larger diameter (< or = 1.9 nm) than the trans opening (< or = 1.6 nm). The channel lumen appeared constricted, with a diameter of < or = 1.2 nm. Cup-shaped lumen geometry was deduced for both channel openings, which appeared to be connected to each other via a central narrow part. The latter contributed significantly to the total electrical resistance and determined the discontinuous character of channel filling with nonelectrolytes. Comparisons of the properties of pores formed by cytolysins of two V. cholerae biotypes (EL Tor and non-O1) indicated that the two ion channels possessed a similar geometry.
authors.affiliation: Laboratory of Membrane Biophysics, Center of Biological Sciences, Department of Biophysics and Radiobiology, Federal University of Pernambuco, Recife, PE, Brazil.
authors.forename: L N
authors.initials: LN
authors.lastname: Yuldasheva
authors.forename: P G
authors.initials: PG
authors.lastname: Merzlyak
authors.forename: A O
authors.initials: AO
authors.lastname: Zitzer
authors.forename: C G
authors.initials: CG
authors.lastname: Rodrigues
authors.forename: S
authors.initials: S
authors.lastname: Bhakdi
authors.forename: O V
authors.initials: OV
authors.lastname: Krasilnikov
mesh_terms: D003603:Cytotoxins / Q000737:chemistry*; D004563:Electrochemistry; D004573:Electrolytes; D007473:Ion Channels / Q000737:chemistry*; D008962:Models, Theoretical; D014734:Vibrio cholerae / Q000737:chemistry* / Q000235:genetics
chemical_list: D003603:Cytotoxins; D004573:Electrolytes; D007473:Ion Channels