Pervanadate mediated an increased generation of inositol phosphates and tension in rat myometrium. Activation and phosphorylation of phospholipase C-gamma 1.
Stimulation of [3H]inositol-labeled rat myometrial strips with pervanadate, formed by mixing orthovanadate and H2O2, induced a dose-dependent accumulation of [3H]inositol phosphates. Orthovanadate or H2O2 added alone had no effect. Pretreatment of myometrium with two tyrosine kinase inhibitors, namely genistein and tyrphostin 47 (at 100 microM), reduced pervanadate-stimulated inositol phosphate formation by 50%. Pervanadate induced a time-sequential formation of inositol phosphates in the order inositol trisphosphate, inositol bisphosphate, and inositol monophosphate. The inhibitory effect of genistein was observed at the level of the three inositol phosphates. Pervanadate induced contraction of the myometrium; the response was dose-dependent. H2O2 or orthovanadate was without effect. Pervanadate-mediated contraction was inhibited (50%) by genistein and tyrphostin 47 (100 microM). Western blot analysis, using anti-phosphotyrosine antibodies, revealed that phosphorylated proteins were present in detergent extracts from pervanadate-stimulated myometrium. Tyrosine phosphorylation was reduced by a preincubation with 100 microM genistein or tyrphostin 47. Phospholipase C-gamma1 was immunodetected in myometrial extracts and was identified as one of the substrates subject to tyrosine phosphorylation following pervanadate treatment. The results demonstrate that, in myometrium, protein tyrosine kinase/phosphatase activities controlled both phosphorylation and activation of phospholipase C-gamma1, contributing to the modulation of the generation of inositol phosphates and tension.
Palmier
B
B
CNRS-URA 1131, Université Paris-Sud, Orsay, France.
Leiber
D
D
Harbon
S
S
D000818:Animals; D015153:Blotting, Western; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D019833:Genistein; D007295:Inositol Phosphates / Q000096:biosynthesis*; D007527:Isoenzymes / Q000378:metabolism*; D007529:Isoflavones / Q000494:pharmacology; D009215:Myometrium / Q000187:drug effects; D009570:Nitriles / Q000494:pharmacology; D010636:Phenols / Q000494:pharmacology; D051966:Phospholipase C gamma; D010766:Phosphorylation; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D010738:Type C Phospholipases / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism; D020032:Tyrphostins; D014590:Uterine Contraction / Q000187:drug effects*; D014638:Vanadates / Q000037:antagonists & inhibitors / Q000494:pharmacology*
D004791:Enzyme Inhibitors; D007295:Inositol Phosphates; D007527:Isoenzymes; D007529:Isoflavones; D009570:Nitriles; D010636:Phenols; D020032:Tyrphostins; C076715:pervanadate; C060152:tyrphostin 47; D014638:Vanadates; D014443:Tyrosine; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D010738:Type C Phospholipases; D051966:Phospholipase C gamma