Stimulation of canine cardiac sarcoplasmic reticulum Ca2+ uptake by dihydropyridine Ca2+ antagonists.
We examined the effects of four Ca2+ antagonists that possess the ability to bind to calmodulin-felodipine, nitrendipine, prenylamine, and verapamil--as well as the effect of the calmodulin antagonist trifluoperazine on Ca2+ uptake and Ca2+ + Mg2+/ATPase activity in canine cardiac sarcoplasmic reticulum. In the presence of 20-30 microM felodipine and 100-200 microM nitrendipine, Ca2+ uptake increased from 69 nmoles X mg-1 X min-1 to 107 and 108 nmoles X mg-1 X min-1, respectively, with half-maximal stimulation occurring at 7.5 and 28 microM respectively. Ca2+ + Mg2+/ATPase activity was unchanged over the same concentration ranges. In contrast, both Ca2+ uptake and Ca2+ + Mg2+/ATPase activities were inhibited in the presence of 10-100 microM trifluoperazine (IC50 = 25 microM), 10-100 microM prenylamine (IC50 = 35 microM) and 100-200 microM verapamil (inhibition insufficient for IC50 determination). None of the drugs affected membrane permeability to Ca2+ as determined by passive 45Ca2+ efflux in the presence of ethyleneglycol bis(beta-amenoethyl ether)N,N,N1-tetraacetic acid (EGTA). Drug inhibition of calmodulin-dependent turkey gizzard myosin light chain kinase activation in a purified protein system was used as a direct measure of calmodulin antagonism, and felodipine, nitrendipine, trifluoperazine, prenylamine, and verapamil blocked this activation at IC50 values of 9.8, 55, 6.4, 31, and 93 microM respectively. None of the drugs studied, however, had any effect upon endogenous phospholamban phosphorylation in our cardiac sarcoplasmic reticulum preparations. These observations indicate that dihydropyridine Ca2+ antagonists stimulate cardiac sarcoplasmic reticulum Ca2+ uptake in vitro either by increasing the efficiency of the transport process or by inhibiting Ca2+-dependent Ca2+ release, and suggest that these effects do not result from interference with calmodulin-mediated processes.
Movsesian
M A
MA
Ambudkar
I S
IS
Adelstein
R S
RS
Shamoo
A E
AE
D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D000252:Calcium-Transporting ATPases / Q000032:analysis; D002147:Calmodulin / Q000037:antagonists & inhibitors; D004095:Dihydropyridines; D004285:Dogs; D066298:In Vitro Techniques; D008297:Male; D009206:Myocardium / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology*; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism*
D002121:Calcium Channel Blockers; D002147:Calmodulin; D004095:Dihydropyridines; D011725:Pyridines; C038806:1,4-dihydropyridine; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases; D002118:Calcium