title: Atomic proximity between S4 segment and pore domain in Shaker potassium channels.
abstract: A recently proposed model for voltage-dependent activation in K+ channels, largely influenced by the KvAP X-ray structure, suggests that S4 is located at the periphery of the channel and moves through the lipid bilayer upon depolarization. To investigate the physical distance between S4 and the pore domain in functional channels in a native membrane environment, we engineered pairs of cysteines, one each in S4 and the pore of Shaker channels, and identified two instances of spontaneous intersubunit disulfide bond formation, between R362C/A419C and R362C/F416C. After reduction, these cysteine pairs bound Cd2+ with high affinity, verifying that the residues are in atomic proximity. Molecular modeling based on the MthK structure revealed a single position for S4 that was consistent with our results and many other experimental constraints. The model predicts that S4 is located in the groove between pore domains from different subunits, rather than at the periphery of the protein.
authors.affiliation: Department of Physiology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California 90095, USA.
authors.forename: Muriel
authors.initials: M
authors.lastname: Lainé
authors.forename: Meng-chin A
authors.initials: MC
authors.lastname: Lin
authors.forename: John P A
authors.initials: JP
authors.lastname: Bannister
authors.forename: William R
authors.initials: WR
authors.lastname: Silverman
authors.forename: Allan F
authors.initials: AF
authors.lastname: Mock
authors.forename: Benoit
authors.initials: B
authors.lastname: Roux
authors.forename: Diane M
authors.initials: DM
authors.lastname: Papazian
mesh_terms: D000818:Animals; D005260:Female; D008564:Membrane Potentials / Q000502:physiology; D008958:Models, Molecular; D009154:Mutation; D009865:Oocytes; D015221:Potassium Channels / Q000737:chemistry* / Q000502:physiology*; D051637:Shaker Superfamily of Potassium Channels; D014981:Xenopus
chemical_list: D015221:Potassium Channels; D051637:Shaker Superfamily of Potassium Channels