Protein methylation activates reconstituted ryanodine receptor-ca release channels from coronary artery myocytes.
Ryanodine receptors (RyR) play an important role in the regulation of intracellular Ca(2+) concentration and in the control of vascular tone. However, the mechanism regulating the activity of RyR is poorly understood. The present study determined whether protein methylation participates in the control of RyR activity. Using a planar lipid bilayer clamping system, S-adenosyl-L-methionine (SAM), a methyl donor, significantly increased the activity of a 245-pS reconstituted Ca(2+) release channel from coronary arterial smooth muscle (CASM) in a concentration-dependent manner. Addition of the protein methylation blockers, 3-deazaadenosine, S-adenosylhomocysteine or sinefungin into the cis solution markedly attenuated SAM-induced activation of RyR/Ca(2+) release channels. By Western blot analysis, arginine N-methyltransferase (PRMT1) and FK506 binding protein (FKBP) were detected in the SR used for reconstitution of RyR. In the presence of anti-PRMT1 antibody (1:100), SAM-induced activation of RyR/Ca(2+) channel was completely abolished. In addition, this SAM-induced increase in RyR/Ca(2+) channel activity was blocked by 30 microM ryanodine and by FK506 (100 microM), a ligand for the RyR accessory protein. These results suggest that protein methylation activates RyR/Ca(2+) release channels and may participate in the control of intracellular Ca(2+) mobilization in CASM cells by transferring a methyl group to the arginine moiety of the RyR accessory protein, FKBP 12.
Chen
Ya-Fei
YF
Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee 53226, USA.
Zhang
Andrew Y
AY
Zou
Ai-Ping
AP
Campbell
William B
WB
Li
Pin-Lan
PL
D000241:Adenosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001158:Arteries / Q000166:cytology / Q000378:metabolism; D002417:Cattle; D003331:Coronary Vessels / Q000166:cytology / Q000378:metabolism*; D008745:Methylation / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D011484:Protein-Arginine N-Methyltransferases / Q000276:immunology; D012433:Ryanodine / Q000494:pharmacology; D019837:Ryanodine Receptor Calcium Release Channel / Q000187:drug effects / Q000378:metabolism*; D012436:S-Adenosylmethionine / Q000494:pharmacology; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D016559:Tacrolimus / Q000494:pharmacology; D022061:Tacrolimus Binding Protein 1A / Q000276:immunology / Q000378:metabolism; D014372:Tubercidin / Q000494:pharmacology
D000906:Antibodies; D019837:Ryanodine Receptor Calcium Release Channel; C018258:3-deazaadenosine; D012433:Ryanodine; D012436:S-Adenosylmethionine; D011484:Protein-Arginine N-Methyltransferases; D022061:Tacrolimus Binding Protein 1A; D000241:Adenosine; D014372:Tubercidin; C006235:sinefungin; D016559:Tacrolimus