muI Na+ channels expressed transiently in human embryonic kidney cells: biochemical and biophysical properties.
We describe the transient expression of the rat skeletal muscle muI Na+ channel in human embryonic kidney (HEK 293) cells. Functional channels appear at a density of approximately 30 in a 10 microns 2 patch, comparable to those of native excitable cells. Unlike muI currents in oocytes, inactivation gating is predominantly (approximately 97%) fast, although clear evidence is provided for noninactivating gating modes, which have been linked to anomalous behavior in the inherited disorder hyperkalemic periodic paralysis. Sequence-specific antibodies detect a approximately 230 kd glycopeptide. The majority of molecules acquire only neutral oligosaccharides and are retained within the cell. Electrophoretic mobility on SDS gels suggests the molecules may acquire covalently attached lipid. The channel is readily phosphorylated by activation of the protein kinase A and protein kinase C second messenger pathways.
Ukomadu
C
C
Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06510.
Zhou
J
J
Sigworth
F J
FJ
Agnew
W S
WS
D000818:Animals; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006023:Glycoproteins / Q000502:physiology; D006031:Glycosylation; D006596:Hexosaminidases / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D015640:Ion Channel Gating; D008970:Molecular Weight; D009132:Muscles / Q000502:physiology; D009439:Neuraminidase / Q000494:pharmacology; D010750:Phosphoproteins / Q000502:physiology; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D015222:Sodium Channels / Q000502:physiology*; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
D006023:Glycoproteins; D010750:Phosphoproteins; D015222:Sodium Channels; D014415:Tunicamycin; D011494:Protein Kinases; D006596:Hexosaminidases; D009439:Neuraminidase