The epsilon-isoform of PKC mediates the hypertonic activation of cation channels in confluent monolayers of rat hepatocytes.
We were interested whether PKC alpha, delta, epsilon or zeta is the isoform actually employed in the activation of hypertonicity-induced cation channels (HICCs) in primary cultures of rat hepatocytes. Quantitative SDS-page and Western-blot experiments revealed that PKC alpha, delta and epsilon were stimulated by Indolactam V (as a DAG substitute for activation of c and nPKCs) but that only PKC delta and epsilon did respond to hypertonic stress. Furthermore, chelation of intracellular Ca(++) by BAPTA-AM did not alter HICC activation in cable-analysis experiments whereas Indolactam V as well as V8 (an Indolactam derivative specific for PKC delta and epsilon) activated HICC currents under isotonic conditions. Finally, by use of Rottlerin (as an inhibitor exhibiting a slight preference for PKC delta over epsilon) PKC epsilon could be identified as the most likely isoform responsible for the activation of the HICC.
Bierhals
Katrin
K
Department of Systemic Cell Biology, Max-Planck-Institute for Molecular Physiology, Dortmund, Germany.
Sondersorg
Anna C
AC
Lin
Chiann-Tso
CT
Rosenbaum
Claudia
C
Waldmann
Herbert
H
Wehner
Frank
F
D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D001578:Benzopyrans / Q000494:pharmacology; D002478:Cells, Cultured; D004789:Enzyme Activation; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D007211:Indoles / Q000494:pharmacology; D007473:Ion Channels / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism; D007769:Lactams / Q000494:pharmacology; D051744:Protein Kinase C-epsilon / Q000378:metabolism*; D051381:Rats
D000098:Acetophenones; D001578:Benzopyrans; D007211:Indoles; D007473:Ion Channels; D007527:Isoenzymes; D007769:Lactams; C043969:indolactam V; C085746:rottlerin; D051744:Protein Kinase C-epsilon