Inhibition of L-type calcium current in rat ventricular cells by the tyrosine kinase inhibitor, genistein and its inactive analog, daidzein.
Effects of genistein, a specific inhibitor of tyrosine kinase, on the L-type Ca2+ channels were examined in freshly isolated young (days 10-18) rat ventricular cells using the whole-cell patch-clamp technique. Bath application of genistein decreased the L-type Ca2+ current [I(Ca(L))] in a concentration-dependent manner. The maximal inhibition of I(Ca(L)) was about 40% (attained at concentrations above 100 microM); the concentration for half-inhibition (IC50) was 11 microM. The effect of genistein (applied for about 5 min) was poorly reversible after washout for up to 5 min. The potential for half-inhibition (Vh) of the steady-state inactivation curve was shifted in the negative direction by 7 mV (at 100 microM) by genistein, and the slope factor was also slightly changed; the activation curve was not affected. Daidzein, which is structurally related to genistein, but has little inhibitory effect on tyrosine kinase activity (of the EGF receptor), unexpectedly had almost the same inhibitory effect on I(Ca(L)). These observations suggest two possibilities for modulation of I(Ca(L)) in rat ventricular cells by genistein: (a) phosphorylation of the slow Ca2+ channels by tyrosine kinase; and (b) direct inhibition of the slow Ca2+ channels (i.e. independent of inhibition of tyrosine kinase activity).
Yokoshiki
H
H
Department of Molecular and Cellular Physiology, College of Medicine, University of Cincinnati, Ohio 45267, USA.
Sumii
K
K
Sperelakis
N
N
D000200:Action Potentials / Q000502:physiology*; D000818:Animals; D015220:Calcium Channels / Q000378:metabolism* / Q000502:physiology*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D019833:Genistein; D006352:Heart Ventricles / Q000166:cytology / Q000378:metabolism*; D066298:In Vitro Techniques; D007529:Isoflavones / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D051381:Rats; D013997:Time Factors; D016276:Ventricular Function
D015220:Calcium Channels; D004791:Enzyme Inhibitors; D007529:Isoflavones; C004742:daidzein; D019833:Genistein; D011505:Protein-Tyrosine Kinases