The cytoplasmic tail of large conductance, voltage- and Ca2+-activated K+ (MaxiK) channel is necessary for its cell surface expression.
The large conductance, voltage- and Ca(2+)-activated K(+) channel (MaxiK) is expressed in several renal segments and functions in cell volume regulation and flow-mediated K(+) secretion. Previously, we cloned two MaxiK channel isoforms, named rbslo1 and rbslo2, from rabbit renal cells. rbslo1 has a 58-amino acid insertion after the S8 hydrophobic domain, whereas rbslo2 is truncated and cannot be activated. Here we use the sequence differences between the two variants to examine their plasma membrane processing. Plasma membrane localization of rbslo1 and 2 expressed in HEK293 cells was assayed by electrophysiology, immunocytochemistry, and biochemistry studies. Consistent with its functional silence, rbslo2 localized primarily within the cytoplasm, presumably in the endoplasmic reticulum and Golgi region. Coexpression with MaxiK beta subunits did not alter the cellular localization of either rbslo1 or rbslo2. When rbslo1 and 2 are cotransfected in non-polarized cells, they colocalized primarily within the cell with only rbslo1 detected at the plasma membrane. When transfected into polarized, medullary-thick ascending limb (mTAL) cells, rbslo1 is expressed at the apical membrane whereas the majority of rbslo2 localized throughout the cytoplasm. Given the high degree of similarity between the two isoforms, we conclude that the cytoplasmic tail of rbslo1 is important for the cell surface expression of MaxiK channels.
Wang
Shao-Xiong
SX
Department of Physiology, School of Medicine, The Johns Hopkins University, Baltimore, Maryland 21205, USA.
Ikeda
Masahiro
M
Guggino
William B
WB
D000818:Animals; D019921:Biotinylation; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004594:Electrophysiology; D006801:Humans; D007150:Immunohistochemistry; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits; D051036:Large-Conductance Calcium-Activated Potassium Channels; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D024681:Potassium Channels, Calcium-Activated / Q000737:chemistry* / Q000378:metabolism*; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D014162:Transfection
D018076:DNA, Complementary; C494841:KCNMA1 protein, human; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits; D051036:Large-Conductance Calcium-Activated Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D020033:Protein Isoforms