Ile-177 and Ser-180 in the S1 segment are critically important in Kv1.1 channel function.
Ile-177 and Ser-180 are conserved residues in the first transmembrane segment (S1) of the Shaker, Shab, Shaw, and Shal subfamilies of voltage-gated K+ channels. Here we report that the mutation of these residues in Kv1.1 to leucine, proline, or arginine abolished the expression of outward potassium currents in Xenopus oocytes. Co-injection of these mutant cRNAs and wild type Kv1.1 cRNA into Xenopus oocytes exerted a potent dominant negative effect resulting in the suppression of Kv1.1-encoded currents. Transient transfection experiments of COS-7 cells revealed that the S1 mutants directed the synthesis of Kv1.1 polypeptides. Quantitative co-immunoprecipitation assays revealed that most of the S1 mutants co-assembled and formed both homo- and heteromultimeric complexes. Furthermore, the mutated polypeptides could reach the plasma membranes of transfected Sol8 cells. We conclude that mutations of Ile-177 and Ser-180 do not interfere with either the assembly of multimeric channel complexes or the targeting of these complexes to the plasma membrane. It is likely that these residues are involved in helix-helix interactions that are critical to the proper functioning of voltage-gated potassium channels.
Mathur
R
R
Cardiovascular Division, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Zhou
J
J
Babila
T
T
Koren
G
G
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D007532:Isoleucine / Q000378:metabolism*; D051662:Kv1.1 Potassium Channel; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D016296:Mutagenesis; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000737:chemistry / Q000378:metabolism*; D024642:Potassium Channels, Voltage-Gated; D051381:Rats; D017386:Sequence Homology, Amino Acid; D012694:Serine / Q000378:metabolism*; D014981:Xenopus
D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D007532:Isoleucine; D051662:Kv1.1 Potassium Channel; D012694:Serine