A myosin phosphatase modulates contractility in skinned smooth muscle.
The influence of a purified holoenzyme form of polycation-modulable (PCM-) myosin phosphatase on Ca2+-dependent actin-myosin interactions was studied in detergent-skinned smooth muscle fibers from chicken gizzard. The concentration of Ca2+ required for half maximal isometric contraction (A0.5; 0.26 microM) of fibers incubated in the absence of phosphatase was increased 2-fold when PCM-phosphatase (13 U/ml) was included in the medium. Removal of the phosphatase restored A0.5 to control level showing that the enzyme-mediated decrease in Ca2+-sensitivity was reversible. Two-dimensional electrophoresis of fiber homogenates revealed that PCM-phosphatase decreased Ca2+-sensitivity for phosphorylation of the regulatory myosin light chains in parallel fashion. Ca2+-dependent increases in isometric force were directly correlated to increases in the extent of light chain phosphorylation up to about 0.35 mol PO4/mol light chain; further increases in phosphorylation were not associated with further increases in force. Addition of PCM-phosphatase to fibers which had been contracted with a suboptimal concentration of Ca2+ (0.35 microM) resulted in rapid relaxation. Unloaded shortening velocity, reflecting cross-bridge cycling rate, was reduced by 92% in the presence of PCM-phosphatase and light chain phosphorylation was decreased by 50%. These data show that both tension and unloaded shortening velocity may be related to Ca2+-dependent phosphorylation of the light chains. The results indicate that the level of phosphorylation attained in the fiber preparations studied probably reflects the ratio of myosin kinase to phosphatase activities. Since protein phosphatases are regulated enzymes the results also suggest that modulation of phosphatase activity may participate in control of smooth muscle contractility.
Bialojan
C
C
II. Physiologisches Institut, Universität Heidelberg, Federal Republic of Germany.
Rüegg
J C
JC
DiSalvo
J
J
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002645:Chickens; D005895:Gizzard, Avian; D007537:Isometric Contraction / Q000187:drug effects; D008297:Male; D009119:Muscle Contraction; D009126:Muscle Relaxation / Q000187:drug effects; D009130:Muscle, Smooth / Q000502:physiology*; D043263:Myosin-Light-Chain Phosphatase; D009218:Myosins / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000502:physiology*; D010766:Phosphorylation
D010749:Phosphoprotein Phosphatases; D043263:Myosin-Light-Chain Phosphatase; D009218:Myosins; D002118:Calcium