Arachidonic acid inhibits myosin light chain phosphatase and sensitizes smooth muscle to calcium.
Arachidonic acid (AA) increased, at constant Ca2+, the levels of force and 20-kDa myosin light chain (MLC20) phosphorylation in permeabilized smooth muscle, and slowed relaxation and MLC20 dephosphorylation. The Ca(2+)-sensitizing effect of AA was not inhibited by inhibitors of AA metabolism (indomethacin, nordihydroguaiaretic acid, or propyl gallate), of protein kinase C (pseudopeptide) or by guanosine-5'-O-(beta-thiodiphosphate) and was abolished by oxidation of AA in air. A non-metabolizable AA analog, 5,8,11,14-eicosatetraynoic acid) also had Ca(2+)-sensitizing effects. Extensive treatment with saponin abolished the Ca(2+)-sensitizing effects of phorbol 12,13-dibutyrate and guanosine-5'-O-(gamma-thiotriphosphate), but not that of AA. A purified, oligomeric MLC20 phosphatase isolated from gizzard smooth muscle was dissociated into subunits by AA, and its activity was inhibited toward heavy meromyosin but not phosphorylase. We conclude that AA may act as a messenger-promoting protein phosphorylation through direct inhibition of the form of protein phosphatase(s) that dephosphorylate MLC20 in vivo.
Gong
M C
MC
Department of Physiology, University of Virginia Medical School, Charlottesville 22908.
Fuglsang
A
A
Alessi
D
D
Kobayashi
S
S
Cohen
P
P
Somlyo
A V
AV
Somlyo
A P
AP
D000818:Animals; D016718:Arachidonic Acid / Q000502:physiology*; D002118:Calcium / Q000378:metabolism*; D005230:Fatty Acids, Nonesterified / Q000502:physiology; D019204:GTP-Binding Proteins / Q000378:metabolism; D066298:In Vitro Techniques; D009126:Muscle Relaxation / Q000502:physiology; D009130:Muscle, Smooth / Q000201:enzymology / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D043263:Myosin-Light-Chain Phosphatase; D019301:Oleic Acid; D009829:Oleic Acids / Q000502:physiology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D011817:Rabbits
D005230:Fatty Acids, Nonesterified; D009829:Oleic Acids; D016718:Arachidonic Acid; D019301:Oleic Acid; D011493:Protein Kinase C; D010749:Phosphoprotein Phosphatases; D043263:Myosin-Light-Chain Phosphatase; D019204:GTP-Binding Proteins; D002118:Calcium