Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells
The L-type Ca 2+ channel is the primary voltage-dependent Ca 2+ -influx pathway in many excitable and secretory cells, and direct phosphorylation by different kinases is one of the mechanisms involved in the regulation of its activity. The aim of this study was to evaluate the participation of Ser/T...
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2007
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| 008 | 190411s2007 xx ||||fo|||| 00| 0 eng|d | ||
| 024 | 7 | |2 scopus |a 2-s2.0-34548719413 | |
| 024 | 7 | |2 cas |a 4 (3 chloroanilino) 6,7 dimethoxyquinazoline, 153436-53-4; calcium ion, 14127-61-8; chelerythrine, 34316-15-9; epidermal growth factor, 62229-50-9; fura 2 acetoxymethyl ester, 105344-37-4, 108964-32-5; genistein, 446-72-0; n [2 (4 bromocinnamylamino)ethyl] 5 isoquinolinesulfonamide, 127243-85-0; nifedipine, 21829-25-4; okadaic acid, 78111-17-8; orthovanadic acid, 14333-18-7; phorbol 13 acetate 12 myristate, 16561-29-8; phosphatase, 9013-05-2; phosphotransferase, 9031-09-8, 9031-44-1; potassium chloride, 7447-40-7; potassium ion, 24203-36-9; protein kinase C, 141436-78-4; protein tyrosine kinase, 80449-02-1; vasculotropin, 127464-60-2; Calcium Channels, L-Type; Calcium, 7440-70-2; Carcinogens; Cyclic AMP-Dependent Protein Kinases, EC 2.7.1.37; Fluorescent Dyes; Fura-2, 96314-98-6; fura-2-am, 105344-37-4; H 89, 127243-85-0; Isoquinolines; Potassium Chloride, 7447-40-7; Protein Kinase C, EC 2.7.1.37; Protein Kinase Inhibitors; Protein Kinases, EC 2.7.1.37; Protein-Serine-Threonine Kinases, EC 2.7.1.37; Protein-Tyrosine Kinases, EC 2.7.1.112; Proto-Oncogene Proteins pp60(c-src), EC 2.7.1.112; Receptor, Epidermal Growth Factor, EC 2.7.1.112; Sulfonamides; Tetradecanoylphorbol Acetate, 16561-29-8 | |
| 040 | |a Scopus |b spa |c AR-BaUEN |d AR-BaUEN | ||
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| 100 | 1 | |a Vela, J. | |
| 245 | 1 | 0 | |a Different kinases regulate activation of voltage-dependent calcium channels by depolarization in GH3 cells |
| 260 | |c 2007 | ||
| 270 | 1 | 0 | |m Díaz-Torga, G.; Instituto de Biología Y Medicina Experimental, CONICET, V. Obligado 2490, (1428) Buenos Aires, Argentina; email: gdiaz@dna.uba.ar |
| 506 | |2 openaire |e Política editorial | ||
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| 520 | 3 | |a The L-type Ca 2+ channel is the primary voltage-dependent Ca 2+ -influx pathway in many excitable and secretory cells, and direct phosphorylation by different kinases is one of the mechanisms involved in the regulation of its activity. The aim of this study was to evaluate the participation of Ser/Thr kinases and tyrosine kinases (TKs) in depolarization-induced Ca 2+ influx in the endocrine somatomammotrope cell line GH3. Intracellular Ca 2+ concentration ([Ca 2+ ] i ) was measured using a spectrofluorometric method with fura 2-AM, and 12.5 mM KCl (K + ) was used as a depolarization stimulus. K + induced an abrupt spike (peak) in [Ca 2+ ] i that was abolished in the presence of nifedipine, showing that K + enhances [Ca 2+ ] i , preferably activating L-type Ca 2+ channels. H89, a selective PKA inhibitor, significantly reduced depolarization-induced Ca 2+ mobilization in a concentration-related manner when it was applied before or after K + , and okadaic acid, an inhibitor of Ser/Thr phosphatases, which has been shown to regulate PKA-stimulated L-type Ca 2+ channels, increased K + -induced Ca 2+ entry. When PKC was activated by PMA, the K + -evoked peak in [Ca 2+ ] i , as well as the plateau phase, was significantly reduced, and chelerythrine (a PKC inhibitor) potentiated the K + -induced increase in [Ca 2+ ] i , indicating an inhibitory role of PKC in voltage-dependent Ca 2+ channel (VDCC) activity. Genistein, a TK inhibitor, reduced the K + -evoked increase in [Ca 2+ ] i , but, unexpectedly, the tyrosine phosphatase inhibitor orthovanadate reduced not only basal Ca 2+ levels but, also, Ca 2+ influx during the plateau phase. Both results suggest that different TKs may act differentially on VDCC activation. Activation of receptor TKs with epidermal growth factor (EGF) or vascular endothelial growth factor potentiated K + -induced Ca 2+ influx, and AG-1478 (an EGF receptor inhibitor) decreased it. However, inhibition of the non-receptor TK pp60 c-Src enhanced K + -induced Ca 2+ influx. The present study strongly demonstrates that a complex equilibrium among different kinases and phosphatases regulates VDCC activity in the pituitary cell line GH3: PKA and receptor TKs, such as vascular endothelial growth factor receptor and EGF receptor, enhance depolarization-induced Ca 2+ influx, whereas PKC and c-Src have an inhibitory effect. These kinases modulate membrane depolarization and may therefore participate in the regulation of a plethora of intracellular processes, such as hormone secretion, gene expression, protein synthesis, and cell proliferation, in pituitary cells. Copyright © 2007 the American Physiological Society. |l eng | |
| 593 | |a Instituto de Biología Y Medicina Experimental, Consejo Nacional de Investigaciones Cientificas Y Tecnicas, Buenos Aires, Argentina | ||
| 593 | |a Instituto de Biología Y Medicina Experimental, CONICET, V. Obligado 2490, (1428) Buenos Aires, Argentina | ||
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