Myosin phosphorylation and Ca2+ sensitization in porcine coronary arterial smooth muscle stimulated with endothelin-1.

We examined the mechanism of endothelin-1-induced contraction in the porcine coronary artery by assessing the changes in myosin light chain phosphorylation and cytosolic Ca2+ concentration ([Ca2+]i) determined by fura-2 and front-surface fluorometry. Endothelin-1 induced a rapid phosphorylation of myosin both in the presence and in the absence of extracellular Ca2+. During the later phase of contraction (5-30 min), the tension and [Ca2+]i were maintained, while the phosphorylated myosin decreased in the presence of extracellular Ca2+. At a similar extent of [Ca2+]i elevation, endothelin-1 induced a greater tension development than did 118 mM K(+)-depolarization and this was not attributed to a greater degree of phosphorylation of myosin. In the Ca(2+)-depleted strips, endothelin-1 induced a slow and transient contraction without an increase in either [Ca2+]i or myosin phosphorylation. Therefore, in addition to the activation of mechanisms of [Ca2+]i-myosin phosphorylation, endothelin-1 may activate additional calcium regulatory mechanisms which may thus increase the Ca2+ sensitivity of contractile elements while also enhance and prolong the contractions in the porcine coronary artery.