title: Salt bridges in the miniature viral channel Kcv are important for function.
abstract: The viral potassium channel Kcv comprises only 94 amino acids, which represent the pore module of more complex K(+) channels. As for Kir-type channels, Kcv also has a short N-terminal helix exposed to the cytoplasm, upstream of the first transmembrane domain. Here we show that this helix is relevant for Kcv function. The presence of charged amino acids, which form dynamic inter- and intra-subunit salt bridges is crucial. Electrophysiological measurements, yeast rescue experiments and molecular dynamics simulations show that mutants in which the critical salt bridge formation is impaired have no or reduced channel activity. We conclude that these salt bridges destabilise the complexation of K(+) ions by negative charges on the inner transmembrane domain at the entrance into the cavity. This feature facilitates a continuous and coordinated transfer of ions between the cavity and the cytoplasm for channels without the canonical bundle crossing.
authors.affiliation: Institute for Botany, TU-Darmstadt, Schnittspahnstrasse 3-5, 64287, Darmstadt, Germany.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Hertel
authors.forename: Sascha
authors.initials: S
authors.lastname: Tayefeh
authors.forename: Thomas
authors.initials: T
authors.lastname: Kloss
authors.forename: Jennifer
authors.initials: J
authors.lastname: Hewing
authors.forename: Manuela
authors.initials: M
authors.lastname: Gebhardt
authors.forename: Dirk
authors.initials: D
authors.lastname: Baumeister
authors.forename: Anna
authors.initials: A
authors.lastname: Moroni
authors.forename: Gerhard
authors.initials: G
authors.lastname: Thiel
authors.forename: Stefan M
authors.initials: SM
authors.lastname: Kast
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D006801:Humans; D008564:Membrane Potentials / Q000502:physiology; D018613:Microscopy, Confocal; D056004:Molecular Dynamics Simulation; D018408:Patch-Clamp Techniques; D017354:Point Mutation; D015221:Potassium Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017433:Protein Structure, Secondary; D012441:Saccharomyces cerevisiae; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D015221:Potassium Channels; D014764:Viral Proteins