Vanadate augments insulin-stimulated insulin receptor kinase activity and prolongs insulin action in rat adipocytes. Evidence for transduction of amplitude of signaling into duration of response.
Vanadate, a protein tyrosine phosphatase inhibitor, preserves insulin-stimulated lipogenesis after removal of insulin. To investigate the mechanism of this action of vanadate, lipogenesis was studied in isolated rat adipocytes exposed to vanadate for 60 min followed by insulin for 15 min at 37 degrees C. Vanadate (10-50 microM) prolonged insulin-stimulated lipogenesis. The half-time (t1/2) of the decay in insulin (0.34 nM)-stimulated lipogenesis after removal of insulin by washing in pH 7.0 followed by pH 7.6 buffer was 21 min in the absence and 59 min in the presence of vanadate. During these conditions, vanadate did not alter insulin binding nor the removal of insulin by the series of washes. In contrast to lipogenesis, the t1/2 of the decay in insulin receptor tyrosine kinase (IRK) activity, assayed with the artificial substrate Poly[Glu:Tyr] (4:1), was not significantly prolonged by vanadate (6 vs. 6.8 min). However, insulin-stimulated IRK activity was markedly augmented by vanadate to 319 +/- 19% of insulin alone, associated with a similar augmentation of phosphotyrosine incorporation into the insulin receptor beta-subunit determined by Western blotting with antiphosphotyrosine antibodies. To determine the relationship between prolongation of lipogenesis and the increase in IRK, adipocytes were exposed to 17.2 nM insulin to activate the IRK to the same extent as insulin (0.34 nM) plus vanadate (maximum activation). During these two conditions, the decay of lipogenesis was similar and after stimulation with 17.2 nM insulin was not prolonged by vanadate. We conclude that vanadate prolongs insulin action at insulin concentrations that do not maximally activate the IRK by augmenting IRK activity.(ABSTRACT TRUNCATED AT 250 WORDS)
Fantus
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Department of Medicine, Royal Victoria Hospital, Montreal, Quebec, Canada.
Ahmad
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Deragon
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D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism; D000818:Animals; D004305:Dose-Response Relationship, Drug; D006207:Half-Life; D007328:Insulin / Q000378:metabolism / Q000494:pharmacology*; D007700:Kinetics; D008055:Lipids / Q000096:biosynthesis; D008297:Male; D019000:Phosphotyrosine; D051381:Rats; D017207:Rats, Sprague-Dawley; D011972:Receptor, Insulin / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D014638:Vanadates / Q000008:administration & dosage / Q000494:pharmacology*
D007328:Insulin; D008055:Lipids; D019000:Phosphotyrosine; D014638:Vanadates; D014443:Tyrosine; D011972:Receptor, Insulin