Published online before print July 8, 2004, doi: 10.1161/01.RES.0000138017.76125.8b
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Submitted on February 25, 2004
Revised on June 18, 2004
Accepted on June 24, 2004
Regulation of Vascular L-type Ca2+ Channels by Phosphatidylinositol 3,4,5-Trisphosphate
Catherine Le Blanc ;
From the Laboratoire de Signalisation et Interactions Cellulaires (C.L.B., C.M., C.B., M.H., N.M.), Université de Bordeaux II, Bordeaux, France; Research Unit "Molecular Cell Biology" (T.B., R.W.), University of Jena, Jena, Germany.
* To whom correspondence should be addressed. E-mail: nathalie.macrez{at}umr5017.u-bordeaux2.fr.
Modulation of voltage-gated L-type Ca2+ channels by phosphoinositide 3-kinase (PI3K) regulates Ca2+ entry and plays a crucial role in vascular excitation-contraction coupling. Angiotensin II (AII) activates Ca2+ entry by stimulating L-type Ca2+ channels through G
-sensitive PI3K in portal vein myocytes. Moreover, PI3K and Ca2+ entry activation have been reported to be necessary for receptor tyrosine kinase-coupled and G protein-coupled receptor-induced DNA synthesis in vascular cells. We have previously shown that tyrosine kinase-regulated class Ia and G protein-regulated class Ib PI3Ks are able to modulate vascular L-type Ca2+ channels. PI3Ks display 2 enzymatic activities: a lipid-kinase activity leading to the formation of phosphatidylinositol 3,4,5-trisphosphate [PI(3,4,5)P3 or PIP3] and a serine-kinase activity. Here we show that exogenous PIP3 applied into the cell through the patch pipette is able to reproduce the Ca2+ channel-stimulating effect of AII and PI3Ks. Moreover, the AII-induced PI3K-mediated stimulation of Ca2+ channel and the resulting increase in cytosolic Ca2+ concentration are blocked by the anti-PIP3 antibody. Mutants of PI3K transfected into vascular myocytes also revealed the essential role of the lipid-kinase activity of PI3K in AII-induced Ca2+ responses. These results suggest that PIP3 is necessary and sufficient to activate a Ca2+ influx in vascular myocytes stimulated by AII.
Key words: L-type Ca2+ channel • PI3K • PIP3 • angiotensin II • smooth muscle
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