title: Calcium ion-mediated regulation of the alpha-toxin pore of Staphylococcus aureus.
abstract: The water-soluble alpha-toxin monomers of Staphylococcus aureus become hexamers forming the transmembrane pore when exposed to the membranes. This pore is freely permeable to small hydrophilic molecules, e.g. carboxyfluorescein, and becomes less permeable in the presence of calcium ions. Calcium ion-mediated decrease of the carboxyfluorescein leakage could not be eliminated by EDTA added in the medium, but the carboxyfluorescein could be freed by EDTA added in the intraliposomal space. This result suggests that the alpha-toxin pore changes its conformation as the calcium ion is bound and that the binding site is exposed to the intraliposomal side of the membrane. The interaction between the alpha-toxin hexamer and 8-anilino-1-naphthalene-sulfonic acid (ANS) was monitored by determining the fluorescence in the presence and absence of calcium chloride. The mean distances between the tryptophan residues of the alpha-toxin hexamer and the bound ANS were calculated to be 1.90 and 1.80 nm in the absence and presence, respectively, of calcium ions. The results showed the calcium ion mediated conformational change of the membrane-embedded alpha-toxin hexamer.
authors.affiliation: Department of Cellular Information Sciences, Tokai University School of Medicine, Isehara, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Tokunaga
authors.forename: T
authors.initials: T
authors.lastname: Nakae
mesh_terms: D000817:Anilino Naphthalenesulfonates / Q000737:chemistry; D001427:Bacterial Toxins / Q000378:metabolism / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D002413:Cations, Divalent; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes; D006460:Hemolysin Proteins / Q000378:metabolism*; D008081:Liposomes; D008566:Membranes / Q000187:drug effects* / Q000378:metabolism; D010539:Permeability; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013211:Staphylococcus aureus / Q000378:metabolism*
chemical_list: D000817:Anilino Naphthalenesulfonates; D001427:Bacterial Toxins; D002413:Cations, Divalent; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006460:Hemolysin Proteins; D008081:Liposomes; C021091:staphylococcal alpha-toxin; C024098:6-carboxyfluorescein; C027132:1-anilino-8-naphthalenesulfonate; D002118:Calcium