Identification of a cytoplasmic domain important in the polarized expression and clustering of the Kv2.1 K+ channel.
The voltage-sensitive K+ channel Kv2.1 has a polarized and clustered distribution in neurons. To investigate the basis for this localization, we expressed wild-type Kv2.1 and two COOH-terminal truncation mutants, delta C318 and delta C187, in polarized epithelial MDCK cells. These functional channel proteins had differing subcellular localization, in that while both wild-type Kv2.1 and delta C187 localized to the lateral membrane in high density clusters, delta C318 was expressed uniformly on both apical and lateral membranes. A chimeric protein containing the hemagglutinin protein from influenza virus and the region of Kv2.1 that differentiates the two truncation mutants (amino acids 536-666) was also expressed in MDCK cells, where it was found in high density clusters similar to those observed for Kv2.1. Polarized expression and clustering of Kv2.1 correlates with detergent solubility, suggesting that interaction with the detergent insoluble cytoskeleton may be necessary for proper localization of this channel.
Scannevin
R H
RH
Department of Biochemistry and Cell Biology, State University of New York, Stony Brook 11794, USA.
Murakoshi
H
H
Rhodes
K J
KJ
Trimmer
J S
JS
D000818:Animals; D001665:Binding Sites; D001710:Biotin / Q000378:metabolism; D001921:Brain / Q000166:cytology; D019556:COS Cells; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D016764:Cell Polarity; D003593:Cytoplasm / Q000378:metabolism; D051668:Delayed Rectifier Potassium Channels; D008297:Male; D016296:Mutagenesis; D009474:Neurons / Q000378:metabolism; D017830:Octoxynol; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism*; D024642:Potassium Channels, Voltage-Gated; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D051669:Shab Potassium Channels; D012995:Solubility
D051668:Delayed Rectifier Potassium Channels; C496991:Kcnb1 protein, rat; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D011993:Recombinant Fusion Proteins; D051669:Shab Potassium Channels; D001710:Biotin; D017830:Octoxynol