FEBS Letters | |
Phospholipase D activation in fibroblast membranes by the α and β isoforms of protein kinase C | |
Exton, John H.1  Conricode, Kevin M.1  Smith, Jennie L.1  Burns, David J.2  | |
[1]Howard Hughes Medical Institute and Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232, USA | |
[2]Sphinx Pharmaceuticals Corp., Durham, NC 27727, USA | |
关键词: Phospholipase D; Protein kinase C; Phosphatidylcholine; Calcium; PLD; phospholipase D; PKC; protein kinase C; PMA; 4β-phorbol 12-myristate 13-acetate; EGTA; [ethylenebis(oxyethylenenitrilo)]tetraacetic acid; HEPES; 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; | |
DOI : 10.1016/0014-5793(94)80490-7 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
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【 摘 要 】
The regulation of phosphatidylcholine-hydrolyzing phospholipase D (PLD) by protein kinase C (PRC) in membranes of Chinese hamster lung fibroblasts (CCL39) was studied using conventional PKC isoforms α, β and γ isolated from rat brain and recombinant PKC isoforms. Cells were incubated with [14C]choline to label endogenous phosphatidylcholine before membranes were prepared and assayed for release of 14C]choline. PKCα was the most potent activator of PLD, producing a maximal effect at approximately 0.1 . PKCβ also stimulated PLD but was less potent and less efficacious, whereas PKCγ was ineffective. Stimulation required addition of a PKC activator, but the isoform specificity was the same whether phorbol 12-myristate 13-acetate (PMA) or Ca2+ was used. Recombinant Ca2+-independent PKC isoforms δ, ϵ, and ζ, failed to stimulate PLD, but recombinant PKCβ1 stimulated PLD in a manner similar to the purified brain PKCβ. Immunoblot analysis of the soluble fraction of CCL 39 fibroblasts detected only the α and ζ isoforms of PKC. The results suggest that PKCα and β are activators of PLD and that PKCα is responsible for the activation in these fibroblasts.
【 授权许可】
Unknown
【 预 览 】
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