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Volume 16, Number 15,
Issue of August 1, 1996
pp. 4617-4624
Copyright ©1996 Society for Neuroscience
Selective G-Protein Regulation of Neuronal Calcium Channels
Received Feb. 15, 1996; revised April 18, 1996; accepted May 2, 1996.
Peter T. Toth1,
Lee R. Shekter1,
Gloria Hui
Ma1,
Louis H. Philipson2, and
Richard J. Miller1
1 Department of Pharmacological and Physiological
Sciences, and 2 Department of Medicine, The University of
Chicago, Chicago, Illinois 60637
We examined the properties and regulation of Ca channels resulting
from the expression of human  1B and
1E subunits stably expressed in HEK293 cells.
The ancillary subunits  1B and
2/ were also stably expressed in these cell
lines. Ca currents in 1B-expressing cells had
the properties of N-type currents. Ca currents in cells expressing
1E exhibited a novel profile that was similar
to the properties of the ``R type'' Ca current. Introduction of
GTP- -S into 1B cells greatly enhanced the
extent of prepulse facilitation of the Ca current, whereas it had only
a very small effect in 1E-expressing cells.
Activation of somatostatin receptors endogenous to HEK293 cells or  opioid receptors, expressed in the cells after transfection, inhibited
Ca currents in 1B-expressing cells. This
inhibition was blocked by pertussis toxin and was partially relieved by
a depolarizing prepulse. In contrast, no inhibitory effects were noted
in cells expressing 1E channels under the same
circumstances. HEK293 cells normally contained G-proteins from all of
the four major families. Inhibition of Ca currents by agonists in
1B-expressing cells was enhanced slightly by
the cotransfection of several G-protein subunits. agonists,
however, had no effect in 1E-containing cells,
even after overexpression of different G-protein -subunits. In
summary, these results demonstrate that there is a large difference in
the susceptibility of 1B- and
1E-based Ca channels to regulation by
G-proteins. This is so despite the fact that the two types of Ca
channels show substantial similarities in their primary sequences.
Key words:
Ca channel;
G-protein;
receptor;
patch
clamp;
HEK293;
somatostatin
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