+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ PDF Full Text
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Translate
+ Recently Requested

Differential translocation of protein kinase C isozymes by thrombin and platelet-derived growth factor. A possible function for phosphatidylcholine-derived diacylglycerol

Differential translocation of protein kinase C isozymes by thrombin and platelet-derived growth factor. A possible function for phosphatidylcholine-derived diacylglycerol

Journal of Biological Chemistry 268(14): 10534-9

The translocation of protein kinase C (PKC) from the cytosolic to the particulate fraction in IIC9 fibroblasts has been studied to define the functions of 1,2-diacylglycerol (DAG) derived from the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP-2) and phosphatidylcholine (PC). alpha-Thrombin caused a biphasic change in DAG, with two peaks at 15-60 s and 5-15 min, derived from PIP-2 and PC, respectively, while platelet-derived growth factor (PDGF) induced a monophasic DAG increase from PC at 5-15 min. alpha-Thrombin also induced a rapid, but transient, increase of inositol 1,4,5-trisphosphate and cytosolic Ca-2+, whereas PDGF did not. Three PKC isozymes, alpha, epsilon, and zeta, were identified by Western blotting in IIC9 cells and were mainly localized in the cytosol. A fraction of cytosolic PKC alpha was rapidly translocated by alpha-thrombin at 15 s, but its membrane association was lost within 1 min. PKC epsilon was also rapidly translocated; however, its membrane association was sustained for almost 60 min. PKC zeta was not translocated by alpha-thrombin or phorbol 12-myristate 13-acetate. PDGF translocated PKC epsilon at 5 min but had little effect at 15 s and did not translocate PKC alpha or zeta. Incubation with Bacillus cereus PC- or phosphatidylinositol-specific phospholipase C, which increased DAG but not phosphatidic acid, stimulated translocation of PKC epsilon, but not PKC alpha or zeta. Addition of chelators to inhibit the rise in intracellular Ca-2+ largely blocked PKC alpha translocation induced by alpha-thrombin but had no effect on PKC epsilon translocation. Addition of ionomycin allowed alpha-thrombin to induce PKC alpha translocation at 5 min. PKC alpha translocation was mimicked by 1,2-dioctanoylglycerol plus ionomycin, but not by either alone. On the other hand, PKC epsilon was translocated by the DAG alone. These results support the conclusion that PIP-2 hydrolysis activates both PKC alpha and epsilon at 15 s, whereas PC hydrolysis activates only PKC epsilon at 5 min. The differential activation at 5 min can be attributed to the failure of PC hydrolysis to increase Ca-2+ and not to a difference in the molecular species of DAG derived from the phospholipids.

(PDF emailed within 1 workday: $29.90)

Accession: 008475786

Download citation: RISBibTeXText

PMID: 8486706

Related references

Dissociation of protein kinase C activation and sn-1,2-diacylglycerol formation. Comparison of phosphatidylinositol- and phosphatidylcholine-derived diglycerides in a-thrombin-stimulated fibroblasts. The Journal of Biological Chemistry 266: 15-21, 1991

Dissociation of protein kinase c activation and sn 1 2 diacylglycerol formation comparison of phosphatidylinositol and phosphatidylcholine derived diglycerides in alpha thrombin stimulated fibroblasts. Journal of Cellular Biochemistry Supplement (15 PART B): 159, 1991

Platelet-derived growth factor-induced p21ras-mediated signaling is independent of platelet-derived growth factor receptor interaction with GTPase-activating protein or phosphatidylinositol-3-kinase. Cell Growth & Differentiation 5(3): 341-347, 1994

Protein kinase C-independent expression of stromelysin by platelet-derived growth factor, ras oncogene, and phosphatidylcholine-hydrolyzing phospholipase C. Journal of Biological Chemistry 266(33): 22597-22602, 1991

Platelet-derived growth factor-BB and thrombin activate phosphoinositide 3-kinase and protein kinase B: role in mediating airway smooth muscle proliferation. Molecular Pharmacology 54(6): 1007-1015, 1998

Protein kinase c in primary glial cell cultures and subcultures 80kda protein phosphorylation the effect of platelet derived growth factor and the turnover of phosphatidylcholine. Levi, G (Ed ) Neurology And Neurobiology, Vol 55 Differentiation And Functions Of Glial Cells; Satellite Meeting Of The International Society For Neurochemistry, Rome, Italy, April 19-21, 1989 Xxviii+429p Wiley-Liss: New York, New York, Usa Illus 87-100, 1990

Platelet-derived growth factor activation of mitogen-activated protein kinase depends on the sequential activation of phosphatidylcholine-specific phospholipase C, protein kinase C-zeta and Raf-1. Biochemical Journal 325: 303-307, 1997

Platelet-derived growth factor activates porcine thecal cell phosphatidylinositol-3-kinase-Akt/PKB and ras-extracellular signal-regulated kinase-1/2 kinase signaling pathways via the platelet-derived growth factor-beta receptor. Endocrinology 141(4): 1545-1553, 2000

Bombesin, platelet-derived growth factor, and diacylglycerol induce selective membrane association and down-regulation of protein kinase C isotypes in Swiss 3T3 cells. Journal of Biological Chemistry 269(4): 2758-2763, 1994

Expression of platelet-derived growth factor beta receptor on human monocyte-derived macrophages and effects of platelet-derived growth factor BB dimer on the cellular function. Journal of Biological Chemistry 268(32): 24353-24360, 1993

The timing and extent of activation of diacylglycerol-responsive protein kinase-Cs determines their ability to inhibit or promote platelet-derived growth factor-dependent DNA synthesis. Experimental Cell Research 281(2): 7-74, 2002

Involvement of phosphatidylcholine-specific phospholipase C in platelet-derived growth factor-induced activation of the mitogen-activated protein kinase pathway in Rat-1 fibroblasts. Journal of Biological Chemistry 272(17): 11011-6, 1997

Expression of platelet-derived growth factor b receptor on human monocyte-derived macrophages and effects of platelet-derived growth factor BB dimer on the cellular function. The Journal of Biological Chemistry 268: 353-60, 1993