Comparison of phospholipase D activity in vasopressin‐ and phorbol ester‐stimulated fibroblasts

FEBS Letters

Wykle, Robert L.¹  Huang, Chunfa¹  Cabot, Myles C.² 

[1] Department of Biochemistry, Bowman Gray School of Medicine, Wake Forest University, Medical Center Boulevard, Winston-Salem, NC 27157-1016, USA;John Wayne Cancer Institute, 2200 Santa Monica Boulevard, Santa Monica, CA 90404, USA

关键词: Phospholipase D;    Vasopressin;    Phorbol ester;    Fibroblast;    REF52;    rat embryo fibroblasts;    TPA;    12-O-tetradecanoylphorbol-13-acetate;    VP;    vasopressin;    PEt;    phosphatidyletha-nol;    PA;    phosphatidic acid;    PLD;    phospholipase D;    PC;    phosphatidylcholine;    PI;    phosphatidylinositol;    DG;    diacylglycerol. In naming of PC;    PI;    PA;    PEt and DG;    we do not distinguish between the type of aliphatic linkage to glycerol (ester;    ether);   

DOI  :  10.1016/0014-5793(93)80054-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Phospholipase D (PLD) activation by vasopressin (VP) was compared to activation by TPA in REF52 cells prelabeled with [³H]glycerol and [¹⁴C]myristic acid. Upon VP-treatment, the formation of [³H] and [¹⁴c]phosphatidic acid (PA) and phosphatidylethanol (PEt) was accompanied by the loss of radioactivity from PC and PI. However, upon TPA-treatment, radioactivity was lost from PC only. No significant changes of phosphatidylethanolamine and phosphatidylserine were detected in the same samples. The inclusion of 5μM staurosporine for 10 min diminished the production of [³H]PEt and [¹⁴C]PEt by 27% and 53% in VP-treated cells, and by 100% and 75% in TPA-treated cells, respectively. Adding 1 mM EGTA to chelate extracellular Ca²⁺ inhibited [³H]PEt by approximately 31% and [¹⁴C]PEt by 17% after VP-stimulation. In contrast, EGTA had no effeet on TPA-stimulation. The data suggest that REF52 cells contain dual PLD activities. The first is stimulated only by VP, requires Ca²⁺ and hydrolyzes PI. The second is stimulated by both TPA and VP, activated by protein kinase C and hydrolyzes PC.

【 授权许可】

Unknown