title: Conformational changes in a mammalian voltage-dependent potassium channel inactivation peptide.
abstract: Fast inactivation is restored in inactivation deletion mutant voltage-gated potassium (Kv) channels by application of synthetic inactivation 'ball' peptide. Using Fourier transform infrared and circular dichroism spectroscopy, we have investigated the structure of synthetic Kv3.4 channel ball peptide, in a range of environments relevant to the function of the ball domain. The ball peptide contains no alpha-helix or beta-sheet in reducing conditions in aqueous solution, but when cosolubilized with anionic lipid or detergent in order to mimic the environment which the ball domain encounters during channel inactivation, the ball peptide adopts a partial beta-sheet structure. Oxidation of the Kv3.4 ball peptide facilitates formation of a disulfide bond between Cys6 and Cys24 and adoption of a partial beta-sheet structure in aqueous solution; the tendency of the oxidized ball peptide to adopt beta-sheet is generally greater than that of the reduced ball peptide in a given environment. THREADER modeling of the Kv3.4 ball peptide structure predicts a beta-hairpin-like conformation which corresponds well to the structure suggested by spectroscopic analysis of the ball peptide in its cyclic arrangement. A V7E mutant Kv3.4 ball peptide analogue of the noninactivating Shaker B L7E mutant ball peptide cannot adopt beta-structure whatever the environment, and regardless of oxidation state. The results suggest that the Kv3.4 ball domain undergoes a conformational change during channel inactivation and may implicate a novel regulatory role for intramolecular disulfide bond formation in the Kv3.4 ball domain in vivo.
authors.affiliation: Department of Biochemistry and Molecular Biology, Royal Free Hospital School of Medicine, London, U.K.
authors.forename: G W
authors.initials: GW
authors.lastname: Abbott
authors.forename: E A
authors.initials: EA
authors.lastname: Mercer
authors.forename: R T
authors.initials: RT
authors.lastname: Miller
authors.forename: B
authors.initials: B
authors.lastname: Ramesh
authors.forename: S K
authors.initials: SK
authors.lastname: Srai
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D002942:Circular Dichroism; D006801:Humans; D015640:Ion Channel Gating; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000037:antagonists & inhibitors* / Q000138:chemical synthesis / Q000235:genetics; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D010455:Peptides; D026902:Potassium Channel Blockers; D015221:Potassium Channels