title: Quantification of group A colicin import sites.
abstract: Pore-forming colicins are soluble bacteriocins which form voltage-gated ion channels in the inner membrane of Escherichia coli. To reach their target, these colicins first bind to a receptor located on the outer membrane and then are translocated through the envelope. Colicins are subdivided into two groups according to the envelope proteins involved in their translocation: group A colicins use the Tol proteins; group B colicins use the proteins TonB, ExbB, and ExbD. We have previously shown that a double-cysteine colicin A mutant which possesses a disulfide bond in its pore-forming domain is translocated through the envelope but is unable to form a channel in the inner membrane (D. Duché, D. Baty, M. Chartier, and L. Letellier, J. Biol. Chem. 269:24820-24825, 1994). Measurements of colicin-induced K+ efflux reveal that preincubation of the cells with the double-cysteine mutant prevents binding of colicins of group A but not of group B. Moreover, we show that the mutant is still in contact with its receptor and import machinery when it interacts with the inner membrane. From these competition experiments, we conclude that each Escherichia coli cell contains approximately 400 and 1,000 colicin A receptors and translocation sites, respectively.
authors.affiliation: Laboratorie d'Ingénierie et de Dynamique des Systèmes Membranaires, Centre National de la Recherche Scientifique, UPR 9027, Marseille, France.
authors.forename: D
authors.initials: D
authors.lastname: Duché
authors.forename: L
authors.initials: L
authors.lastname: Letellier
authors.forename: V
authors.initials: V
authors.lastname: Géli
authors.forename: H
authors.initials: H
authors.lastname: Bénédetti
authors.forename: D
authors.initials: D
authors.lastname: Baty
mesh_terms: D001692:Biological Transport / Q000235:genetics; D003087:Colicins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins; D007473:Ion Channels / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D011188:Potassium / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000032:analysis*
chemical_list: D003087:Colicins; D029968:Escherichia coli Proteins; D007473:Ion Channels; D011956:Receptors, Cell Surface; C028593:colicin receptor, E coli; D011188:Potassium