Dual regulation of the cardiac L-type calcium channel in L6 cells by protein kinase C.
The role of protein kinase C (PKC) in the regulation of cardiac L-type Ca(2+) channel activity (LCC) was investigated in L6 rat neonatal myoblasts. Depolarization of fura-2 loaded cells with 140mM KCl activated a Ba(2+) influx pathway that was blocked by nifedipine and stimulated by (-) Bay K 8644. At least two splice variants of the alpha(1C) subunit of the cardiac LCC were identified by PCR; the alpha(1S) subunit of the skeletal muscle LCC was not detected. Peptides that specifically inhibit translocation of the novel, Ca(2+)-independent delta and epsilon PKC isozymes reduced Ba(2+) influx by 27% and 19%, respectively, whereas a corresponding peptide directed against translocation of classical PKC alpha had no effect. Ingenol 3,20-dibenzoate, an agent reported to selectively activate novel PKCs, increased Ba(2+) uptake by 31% while ethanol, a PKC epsilon agonist, enhanced uptake by 38%. In contrast, selective activation of classical PKCs with thymeleatoxin or an agonist peptide reduced Ba(2+) influx by 23-33%. Ba(2+) influx was reduced by 30-40% when cells were treated with either a PKC inhibitor (Gö 6983, bisindolylmaleimide) or the PKC activator phorbol-12-myristate-13-acetate. We propose that novel, Ca(2+)-insensitive PKC(s) enhance cardiac Ca(2+) channel activity in L6 cells under basal conditions while activation of the classical, Ca(2+)-sensitive PKC(s) inhibits channel activity. These findings provide the first evidence that different PKC isozymes exert class-specific opposing effects on cardiac L-type Ca(2+) channel activity in L6 myoblasts.
Tafti
Bashir A
BA
Stanford University School of Medicine, Stanford, CA 94305, United States.
Hantash
Basil M
BM
D000818:Animals; D020746:Calcium Channels, L-Type / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004789:Enzyme Activation / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D007527:Isoenzymes / Q000378:metabolism; D008954:Models, Biological; D032446:Myoblasts / Q000187:drug effects / Q000201:enzymology; D010455:Peptides / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
D020746:Calcium Channels, L-Type; D007527:Isoenzymes; D010455:Peptides; D000431:Ethanol; C115197:calcium-independent protein kinase C; D011493:Protein Kinase C