Protein kinase C increases force and slows relaxation in smooth muscle: evidence for regulation of the myosin light chain phosphatase.
To determine if activation of protein kinase C (PKC) participates in the molecular mechanism for agonist induced force enhancement, force was measured in single beta-escin skinned smooth muscle cells stimulated to contract with Ca2+, myosin light chain (MLC) kinase, PKC and microcystin-LR. The constituently active fragment of protein kinase C (PKM) increased both force and MLC phosphorylation in cells previously stimulated to contract at submaximal Ca2+. For cells contracted with saturating Ca2+, PKM stimulation did not increase either force or MLC phosphorylation. For contractions stimulated with both PKM and microcystin-LR, force rose significantly slower than contractions produced by Ca2+ or MLC kinase, suggesting that PKM increases force by a decrease in the rate of myosin dephosphorylation. Consistent with this hypothesis is the finding that the rate of force relaxation was slowed by PKM. This is the first direct demonstration that activation of PKC increases force in smooth muscle, and these results suggest that in smooth muscle, agonist induced activation of PKC plays a role in force regulation via an inhibition of myosin light chain phosphatase activity.
Ikebe
M
M
Department of Medicine, Case Western Reserve University, Cleveland, Ohio 44106, USA.
Brozovich
F V
FV
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D004789:Enzyme Activation; D066298:In Vitro Techniques; D008387:Marine Toxins; D052998:Microcystins; D009119:Muscle Contraction / Q000187:drug effects; D009126:Muscle Relaxation / Q000502:physiology; D009131:Muscle, Smooth, Vascular / Q000201:enzymology / Q000502:physiology*; D043263:Myosin-Light-Chain Phosphatase; D010456:Peptides, Cyclic / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D011817:Rabbits
D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D011493:Protein Kinase C; D010749:Phosphoprotein Phosphatases; D043263:Myosin-Light-Chain Phosphatase; C057862:cyanoginosin LR; D002118:Calcium