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J Biol Chem, Vol. 274, Issue 37, 26127-26134, September 10, 1999

Phospholipase C-delta 1 Is Activated by Capacitative Calcium Entry That Follows Phospholipase C-beta Activation upon Bradykinin Stimulation

Yong-Hyun KimDagger , Tae-Ju ParkDagger , Young Han LeeDagger , Kwang Jin Baek§, Pann-Ghill SuhDagger , Sung Ho RyuDagger , and Kyong-Tai KimDagger

From the Dagger  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

To characterize the regulatory mechanism of phospholipase C-delta 1 (PLC-delta 1) in the bradykinin (BK) receptor-mediated signaling pathway, we used a clone of PC12 cells, which stably overexpress PLC-delta 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-delta 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-delta 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-delta 1 is activated by capacitative calcium entry following the activation of PLC-beta , additively inducing IP3 production and Ca2+ rise in BK-stimulated PC12 cells.


Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.



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