Effects of a Shaker K+ channel peptide and trypsin on a K+ channel in Necturus enterocytes.
We have previously demonstrated that a synthetic peptide composed of the first 22 amino acids from the NH2-terminus of the Shaker B K+ channel protein deactivates a voltage-dependent K+ channel present in basolateral membrane of Necturus small intestinal epithelial cells reconstituted into planar lipid bilayers (Dubinsky et al. Proc. Natl. Acad. Sci. USA 89: 1770-1774, 1992). We now demonstrate that this peptide interacts with the inner surface of the Necturus channel only when it is in the open or conducting configuration and that this interaction is hindered by tetraethylammonium ion, a well-established blocker of this and other K+ channels. We conclude that this peptide is an open-pore blocker of the Necturus K+ channel as it appears to be in the case of the Shaker B K+ channel. We further demonstrate that trypsin, which abolishes the ability of this peptide to block both the Necturus and the Shaker K+ channels and inhibits spontaneous inactivation of the Shaker K+ channel, also impairs the voltage-gate of the Necturus K+ channel. These findings, and others to be reported in a companion paper, suggest structural homologies between the "inactivation peptide" of the Shaker B K+ channel and the voltage-gate of the Necturus K+ channel.
Mayorga-Wark
O
O
Department of Physiology and Cell Biology, University of Texas Medical School, Houston 77225.
Costantin
J
J
Dubinsky
W P
WP
Schultz
S G
SG
D000818:Animals; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D004594:Electrophysiology; D007074:Immunoglobulin G / Q000494:pharmacology; D007421:Intestine, Small / Q000166:cytology / Q000378:metabolism*; D009154:Mutation; D009338:Necturus maculosus / Q000378:metabolism*; D010446:Peptide Fragments / Q000494:pharmacology*; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D019789:Tetraethylammonium; D013757:Tetraethylammonium Compounds / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology*
D007074:Immunoglobulin G; D010446:Peptide Fragments; D015221:Potassium Channels; D013757:Tetraethylammonium Compounds; D019789:Tetraethylammonium; D014357:Trypsin