Inhibition of renin secretion by Ca2+ through activation of myosin light chain kinase.
This study sought to identify specific enzyme(s) involved in the biochemical cascade of inhibition of renin secretion through Ca(2+)-calmodulin mediation with the use of inhibitors of protein kinase and phosphatases. Inhibition of renin secretion mediated by Ca(2+)-calmodulin was induced by incubating rat renal cortical slices in K(+)-rich depolarizing medium, producing > 50% inhibition. This inhibition was completely blocked by the calmodulin antagonist calmidazolium. The inhibitor of protein kinase with broad specificity, K-252a, blocked the inhibition of renin secretion. Neither KN-62, a specific inhibitor of Ca(2+)-calmodulin-dependent protein kinase II (CaMK II), nor specific inhibitors of protein phosphatase 2B (PP2B), cyclosporin A and FK-506, blocked the inhibition. On the other hand, all four known inhibitors specific for myosin light chain kinase (MLCK), with different chemical structures and mechanisms of inhibition (ML-9, ML-7, KT-5926 and wortmannin), almost completely protected renin secretion against the inhibition by Ca2+. In particular, ML-9 reversively protected > 77% secretion against the inhibition both in K(+)-rich medium alone and in combination with the calcium ionophore A-23187 in a concentration-dependent manner. Together, these findings from our present study provide the first evidence, albeit indirect in nature, for the possibility that activation of Ca(2+)-calmodulin-dependent MLCK at the downstream of Ca2+ influx into juxtaglomerular (JG) cells leads to phosphorylation of 20-kDa regulatory myosin light chain (MLC20). Through interaction with actin, the phosphorylated MLC20 may play an important role in the inhibitory stimulus-secretion coupling of renin.
Park
C S
CS
Department of Physiology, University of Ulsan College of Medicine, Seoul, Korea.
Chang
S H
SH
Lee
H S
HS
Kim
S H
SH
Chang
J W
JW
Hong
C D
CD
D000818:Animals; D001381:Azepines / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000502:physiology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D003470:Culture Media; D046508:Culture Techniques; D004594:Electrophysiology; D004789:Enzyme Activation; D005260:Female; D007672:Kidney Cortex / Q000378:metabolism / Q000502:physiology; D008297:Male; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism*; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012083:Renin / Q000378:metabolism; D012964:Sodium / Q000494:pharmacology; D000092502:Renin Inhibitors
D001381:Azepines; D000001:Calcimycin; D002118:Calcium; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D003470:Culture Media; D009219:Myosin-Light-Chain Kinase; D011188:Potassium; D012083:Renin; D012964:Sodium; D000092502:Renin Inhibitors; C056218:ML 9