ARF1‐regulated phospholipase D in human neutrophils is enhanced by PMA and MgATP

FEBS Letters

Cronin, Paul¹  Whatmore, Jacqueline¹  Cockcroft, Shamshad¹ 

[1] Department of Physiology, Rockefeller Building, University College London, University Street, London, WC1E 6JJ, UK

关键词: Phospholipase D;    Neutrophil;    ADP ribosylating factor;    PMA;    MgATP;    PLD;    phospholipase D;    PI 3-kinase;    phosphoinositide 3-kinase;    PA;    phosphatidic acid;    PEt;    phosphatidylethanol;    PMA;    phorbol 12-myristate 13-acetate;    PKC;    protein kinase C;    ARF;    ADP-ribosylation factor;    G-proteins;    GTP-binding proteins;    fMetLeuPhe;    N-formyl-methionyl-leucyl-phenylalanine;    GTP-γ-S;    guanosine 5′[γ-thio]triphosphate;    PLC;    inositol lipid-specific phospholipase;    PI;    phosphatidylinositol;    PC;    phosphatidylcholine;   

DOI  :  10.1016/0014-5793(94)00930-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Human neutrophil PLD activity stimulated with GTP-γ-S was reconstituted with recombinant ARF1 in cytosol-depleted cells. PMA-pretreatment of intact cells greatly enhanced the subsequent reconstitution of the ARF1-regulated PLD activity. This enhancement was only observed provided that the intact cells were pretreated with PMA, suggesting the stable recruitment of a cytosolic component, presumably protein kinase C, to the membranes. rARF1-reconstituted PLD activity was not dependent on MGATP, but could be considerably enhanced by MgATP. Maximal effects of MgATP were seen at 1 mM. This enhancement by MgATP could not be attributed to protein kinase C. Neomycin was found to inhibit ARF1-regulated PLD activity suggesting the requirement for polyphosphoinositides. We conclude: (i) that many of the observed effects of PMA may be dependent on the presence of the small GTP-binding protein, ARF, and (ii) polyphosphoinositides are required for ARF1-stimulated PLD activity.

【 授权许可】

Unknown