Caldesmon and heat shock protein 20 phosphorylation in nitroglycerin- and magnesium-induced relaxation of swine carotid artery.
Nitrovasodilators, high extracellular Mg(2+), and some other relaxing agents can cause smooth muscle relaxation without reductions in myosin regulatory light chain (MRLC) phosphorylation. Relaxations without MRLC dephosphorylation suggest that other regulatory systems, beyond MRLC phosphorylation, are present in smooth muscle. We tested whether changes in caldesmon phosphorylation, heat shock protein 20 (HSP20) phosphorylation, or intracellular pH (pH(i)) could be responsible for relaxation without MRLC dephosphorylation. In unstimulated tissues, caldesmon was phosphorylated 1.02+/-0.10 mol P(i)/mol caldesmon (mean+/-1 S.E.M.), HSP20 was phosphorylated 0.005+/-0.003 mol P(i)/mol HSP20, and estimated pH(i) was 7.21+/-0.07. Histamine stimulation induced a contraction, an intracellular acidosis, but did not significantly change caldesmon or HSP20 phosphorylation. Addition of nitroglycerin induced a relaxation, significantly increased HSP20 phosphorylation to 0.18+/-0.02 mol P(i)/mol HSP20, did not significantly change caldesmon phosphorylation, and pH(i) returned to near unstimulated values. Increase in extracellular Mg(2+) to 10 mM induced a relaxation, but did not significantly change HSP20 or caldesmon phosphorylation. These data suggest that changes in caldesmon phosphorylation, HSP20 phosphorylation, or pH(i) cannot be the sole explanation for relaxation without MRLC dephosphorylation. However, it is possible that HSP20 phosphorylation may be involved in nitroglycerin-induced relaxation without MRLC dephosphorylation.
Rembold
C M
CM
Cardiovascular Division, Departments of Internal Medicine and Physiology, University of Virginia Health System, Charlottesville, VA 22908, USA. crembold@virginia.edu
O'Connor
M
M
D000818:Animals; D002148:Calmodulin-Binding Proteins / Q000737:chemistry / Q000494:pharmacology*; D002339:Carotid Arteries / Q000187:drug effects*; D050886:HSP20 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000494:pharmacology*; D006632:Histamine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008274:Magnesium / Q000494:pharmacology*; D009126:Muscle Relaxation / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000187:drug effects; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D005996:Nitroglycerin / Q000494:pharmacology*; D010750:Phosphoproteins / Q000494:pharmacology*; D010766:Phosphorylation; D013552:Swine
D002148:Calmodulin-Binding Proteins; D050886:HSP20 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D010750:Phosphoproteins; D006632:Histamine; D009219:Myosin-Light-Chain Kinase; D005996:Nitroglycerin; D008274:Magnesium