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J Biol Chem, Vol. 274, Issue 37, 26127-26134, September 10, 1999
1 Is Activated by Capacitative Calcium
Entry That Follows Phospholipase C-
Activation upon Bradykinin
Stimulation
,
,
,
,
, and
From the To characterize the regulatory mechanism of
phospholipase C-
Department of Life Science, Pohang
University of Science and Technology, Pohang, 790-784, Republic of
Korea and the § Department of Biochemistry, College of
Medicine, Chung-Ang University, Seoul,
156-756, Republic of Korea
1 (PLC-
1) in the bradykinin (BK)
receptor-mediated signaling pathway, we used a clone of PC12 cells,
which stably overexpress PLC-
1 (PC12-D1). Stimulation with BK
induced a significantly higher Ca2+ elevation and
inositol 1,4,5-trisphosphate (IP3) production with a much
lower half-maximal effective concentration (EC50) of BK in
PC12-D1 cells than in wild type (PC12-W) or vector-transfected (PC12-V)
cells. However, BK-induced intracellular Ca2+ release and
IP3 generation was similar between PC12-V and PC12-D1 cells
in the absence of extracellular Ca2+, suggesting that the
availability of extracellular Ca2+ is essential to the
activation of PLC-
1. When PC12-D1 cells were treated with agents
that induce Ca2+ influx, more IP3 was produced,
suggesting that the Ca2+ entry induces IP3
production in PC12-D1 cells. Furthermore, the additional
IP3 production after BK-induced capacitative calcium entry
was detected in PC12-D1 cells, suggesting that PLC-
1 is mainly
activated by capacitative calcium entry. When cells were stimulated
with BK in the presence of extracellular Ca2+,
[3H]norepinephrine secretion was much greater from
PC12-D1 cells than from PC12-V cells. Our results suggest that PLC-
1
is activated by capacitative calcium entry following the activation of
PLC-
, additively inducing IP3 production and
Ca2+ rise in BK-stimulated PC12 cells.
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