Activation of neuroendocrine L-type channels (alpha1D subunits) in retinal pigment epithelial cells and brain neurons by pp60(c-src).
The aim of this study is to characterize the subtype of tyrosine kinase-regulated L-type Ca(2+) channels in retinal pigment epithelial (RPE) cells. Ca(2+) channel alpha1D-subunits were enriched by immunoprecipitation from membrane proteins isolated from rat RPE cells. Western blot analysis of the precipitates revealed coprecipitation of pp60(c-src). In addition, in precipitates obtained with antibodies against pp60(c-src), alpha1D-subunits were identified. The same was observed in immunoprecipitations from rat brain neurons. Tyrosine phosphorylation of alpha1D-subunits was confirmed using anti-phosphotyrosine antibodies. Ba(2+) currents through L-type channels in cultured rat RPE cells were increased by intracellular application of active pp60(c-src) (30 U/ml) (heat-inactivated pp60(c-src) had no effect). Thus, L-type channels of the neuroendocrine subtype can be expressed in epithelial cells and are activated by tyrosine kinase of the src subtype. This kind of regulation is also suggested for brain-derived neurons.
Strauss
O
O
Institut für Klinische Physiologie, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, Hindenburgdamm 30, Berlin, 12200, Germany. strauss@ukbf.fu-berlin.de
Buss
F
F
Rosenthal
R
R
Fischer
D
D
Mergler
S
S
Stumpff
F
F
Thieme
H
H
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001921:Brain / Q000502:physiology*; D020746:Calcium Channels, L-Type / Q000302:isolation & purification / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D008564:Membrane Potentials / Q000502:physiology; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D009474:Neurons / Q000502:physiology*; D018408:Patch-Clamp Techniques; D010857:Pigment Epithelium of Eye / Q000502:physiology*; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats
D000906:Antibodies; D020746:Calcium Channels, L-Type; D008565:Membrane Proteins; D016392:Proto-Oncogene Proteins pp60(c-src)