title: Structural studies of the N-terminus of Connexin 32 using 1H NMR spectroscopy.
abstract: The amino terminus of gap junction proteins, connexins, plays a fundamental role in voltage gating and ion permeation. We have previously shown with (1)H NMR that the structure of the N-terminus of a representative connexin molecule contains a flexible turn around glycine 12 [P.E. Purnick, D.C. Benjamin, V.K. Verselis, T.A. Bargiello, T.L. Dowd, Arch. Biochem. Biophys. 381 (2000) 181-190] allowing the N-terminus to reside at the cytoplasmic entry of the channel forming a voltage-sensor. Previous functional studies or neuropathies have shown that the mutation G12Y and G12S form non-functional channels while functional channels are formed from G12P. Using 2D (1)H NMR we show that similar to G12, the structure of the G12P mutant contains a more flexible turn around residue 12, whereas the G12S and G12Y mutants contain tighter, helical turns in this region. These results suggest an unconstrained turn is required around residue 12 to position the N-terminus within the pore allowing the formation of the cytoplasmic channel vestibule, which appears to be critical for proper channel function.
authors.affiliation: Department of Chemistry, Brooklyn College of the City University of New York, Brooklyn, NY 11210, USA.
authors.forename: B D
authors.initials: BD
authors.lastname: Kalmatsky
authors.forename: S
authors.initials: S
authors.lastname: Bhagan
authors.forename: Q
authors.initials: Q
authors.lastname: Tang
authors.forename: T A
authors.initials: TA
authors.lastname: Bargiello
authors.forename: T L
authors.initials: TL
authors.lastname: Dowd
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017630:Connexins / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D057927:Hydrophobic and Hydrophilic Interactions; D015640:Ion Channel Gating / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D017433:Protein Structure, Secondary / Q000235:genetics; D000097002:Gap Junction beta-1 Protein
chemical_list: D017630:Connexins; D000097002:Gap Junction beta-1 Protein