title: The pattern of disulfide linkages in the extracellular loop regions of connexin 32 suggests a model for the docking interface of gap junctions.
abstract: Connexins, like true cell adhesion molecules, have extracellular domains that provide strong and specific homophilic, and in some cases, heterophilic interactions between cells. Though the structure of the binding domains of adhesion proteins have been determined, the extracellular domains of connexins, consisting of two loops of approximately 34-37 amino acids each, are not easily studied in isolation from the rest of the molecule. As an alternative, we used a novel application of site-directed mutagenesis in which four of the six conserved cysteines in the extracellular loops of connexin 32 were moved individually and in all possible pairwise and some quadruple combinations. This mapping allowed us to deduce that all disulfides form between the two loops of a single connexin, with the first cysteine in one loop connected to the third of the other. Furthermore, the periodicity of movements that produced functional channels indicated that these loops are likely to form antiparallel beta sheets. A possible model that could explain how these domains from apposed connexins interact to form a complete channel is discussed.
authors.affiliation: Department of Biological Sciences, State University of New York at Buffalo, Buffalo, New York 14260-1300, USA.
authors.forename: C I
authors.initials: CI
authors.lastname: Foote
authors.forename: L
authors.initials: L
authors.lastname: Zhou
authors.forename: X
authors.initials: X
authors.lastname: Zhu
authors.forename: B J
authors.initials: BJ
authors.lastname: Nicholson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D017630:Connexins / Q000235:genetics / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism*; D005110:Extracellular Space; D017629:Gap Junctions / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011817:Rabbits; D014981:Xenopus; D000097002:Gap Junction beta-1 Protein
chemical_list: D017630:Connexins; D004220:Disulfides; D000097002:Gap Junction beta-1 Protein