title: Identification of a pore lining segment in gap junction hemichannels.
abstract: The ability of certain connexins to form open hemichannels has been exploited to study the pore structure of gap junction (hemi)channels. Cysteine scanning mutagenesis was applied to cx46 and to a chimeric connexin, cx32E(1)43, which both form patent hemichannels when expressed in Xenopus oocytes. The thiol reagent maleimido-butyryl-biocytin was used to probe 12 cysteine replacement mutants in the first transmembrane segment and two in the amino-terminal segment. Maleimido-butyryl-biocytin was found to inhibit channel activity with cysteines in two equivalent positions in both connexins: I33C and M34C in cx32E(1)43 and I34C and L35C in cx46. These two positions in the first transmembrane segment are thus accessible from the extracellular space and consequently appear to contribute to the pore lining. The data also suggest that the pore structure is complex and may involve more than one transmembrane segment.
authors.affiliation: Department of Physiology and Biophysics, University of Miami School of Medicine, Florida 33101, USA.
authors.forename: X W
authors.initials: XW
authors.lastname: Zhou
authors.forename: A
authors.initials: A
authors.lastname: Pfahnl
authors.forename: R
authors.initials: R
authors.lastname: Werner
authors.forename: A
authors.initials: A
authors.lastname: Hudder
authors.forename: A
authors.initials: A
authors.lastname: Llanes
authors.forename: A
authors.initials: A
authors.lastname: Luebke
authors.forename: G
authors.initials: G
authors.lastname: Dahl
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017630:Connexins / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry*; D017629:Gap Junctions / Q000737:chemistry*; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry; D008301:Maleimides / Q000737:chemistry*; D016296:Mutagenesis; D009865:Oocytes; D018272:Porins / Q000737:chemistry*; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D014981:Xenopus
chemical_list: D017630:Connexins; D008301:Maleimides; D018272:Porins; C013411:biocytin; D008239:Lysine; D003545:Cysteine