title: A novel N14Y mutation in Connexin26 in keratitis-ichthyosis-deafness syndrome: analyses of altered gap junctional communication and molecular structure of N terminus of mutated Connexin26.
abstract: Connexins (Cxs) are transmembranous proteins that connect adjacent cells via channels known as gap junctions. The N-terminal 21 amino acids of Cx26 are located at the cytoplasmic side of the channel pore and are thought to be essential for the regulation of channel selectivity. We have found a novel mutation, N14Y, in the N-terminal domain of Cx26 in a case of keratitis-ichthyosis-deafness syndrome. Reduced gap junctional intercellular communication was observed in the patient's keratinocytes by the dye transfer assay using scrape-loading methods. The effect of this mutation on molecular structure was investigated using synthetic N-terminal peptides from both wild-type and mutated Cx26. Two-dimensional (1)H nuclear magnetic resonance and circular dichroism measurements demonstrated that the secondary structures of these two model peptides are similar to each other. However, several novel nuclear Overhauser effect signals appeared in the N14Y mutant, and the secondary structure of the mutant peptide was more susceptible to induction of 2,2,2-trifluoroethanol than wild type. Thus, it is likely that the N14Y mutation induces a change in local structural flexibility of the N-terminal domain, which is important for exerting the activity of the channel function, resulting in impaired gap junctional intercellular communication.
authors.affiliation: Department of Dermatology, Hokkaido University Graduate School of Medicine, North 15 West 7, Kita-ku, Sapporo 060-8638, Japan. ariken@med.hokudai.ac.jp
authors.forename: Ken
authors.initials: K
authors.lastname: Arita
authors.forename: Masashi
authors.initials: M
authors.lastname: Akiyama
authors.forename: Tomoyasu
authors.initials: T
authors.lastname: Aizawa
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Umetsu
authors.forename: Ikuo
authors.initials: I
authors.lastname: Segawa
authors.forename: Maki
authors.initials: M
authors.lastname: Goto
authors.forename: Daisuke
authors.initials: D
authors.lastname: Sawamura
authors.forename: Makoto
authors.initials: M
authors.lastname: Demura
authors.forename: Keiichi
authors.initials: K
authors.lastname: Kawano
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shimizu
mesh_terms: D016475:3T3 Cells; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002675:Child, Preschool; D002942:Circular Dichroism; D000072259:Connexin 26; D017630:Connexins / Q000737:chemistry / Q000235:genetics*; D004252:DNA Mutational Analysis; D003638:Deafness / Q000235:genetics*; D005260:Female; D017629:Gap Junctions / Q000473:pathology* / Q000648:ultrastructure; D006801:Humans; D007057:Ichthyosis / Q000235:genetics*; D015603:Keratinocytes / Q000166:cytology / Q000473:pathology; D007634:Keratitis / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry* / Q000235:genetics; D009154:Mutation / Q000235:genetics*; D019906:Nuclear Magnetic Resonance, Biomolecular; D012867:Skin / Q000166:cytology / Q000473:pathology / Q000648:ultrastructure; D013577:Syndrome
chemical_list: D000596:Amino Acids; D017630:Connexins; C000607042:GJB2 protein, human; D050505:Mutant Proteins; D000072259:Connexin 26