Activation of myosin light chain phosphatase in intact arterial smooth muscle during nitric oxide-induced relaxation.
We investigated whether myosin light chain phosphatase activity changes during nitric oxide-induced relaxation of contracted intact carotid media and how changes in phosphatase activity mediate this relaxation. We also investigated one mechanism for regulating this phosphatase. Myosin phosphatase activity, myosin light chain phosphorylation, guanosine 3',5'-cyclic monophosphate (cGMP) concentration, and phosphorylation of the inhibitory protein CPI-17 were all assayed in homogenates of one carotid media ring at each time point during nitric oxide-induced relaxation. The application of sodium nitroprusside to histamine-contracted media caused rapid declines in light chain phosphorylation and force. These were temporally correlated with a rapid elevation of cGMP and a large transient increase in myosin phosphatase activity. During the early response to nitroprusside, when force declined, increases in myosin phosphatase activity, concurrent with cGMP-mediated decreases in calcium and myosin light chain kinase activity, could accelerate light chain dephosphorylation. CPI-17 was dephosphorylated upon application of nitroprusside at the same time that myosin phosphatase activity increased, suggesting that the removal of inhibition by phospho-CPI-17 contributed to the increase in myosin phosphatase activity. After 20 min of nitroprusside, myosin phosphatase activity had declined to basal levels, however low force was sustained. Additional light chain phosphorylation-independent mechanisms may be involved in sustaining the relaxation.
Etter
E F
EF
Department of Molecular Physiology and Biological Physics and the Center for Cell Signaling, University of Virginia School of Medicine, Charlottesville, Virginia 22908-0577, USA. efe3c@virginia.edu
Eto
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Wardle
R L
RL
Brautigan
D L
DL
Murphy
R A
RA
D000818:Animals; D001158:Arteries / Q000201:enzymology* / Q000502:physiology; D006152:Cyclic GMP / Q000502:physiology; D004789:Enzyme Activation; D009124:Muscle Proteins / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000201:enzymology* / Q000502:physiology; D043263:Myosin-Light-Chain Phosphatase; D009569:Nitric Oxide / Q000502:physiology*; D009599:Nitroprusside / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D013552:Swine; D014664:Vasodilation
D009124:Muscle Proteins; D010750:Phosphoproteins; D009599:Nitroprusside; D009569:Nitric Oxide; D010749:Phosphoprotein Phosphatases; D043263:Myosin-Light-Chain Phosphatase; D006152:Cyclic GMP