ATP-dependent activation of phospholipase C by antigen, NECA, Na3VO4, and GTP-gamma-S in permeabilized RBL cell ghosts: differential augmentation by ATP, phosphoenolpyruvate and phosphocreatine.
Ghosts prepared from rat basophilic leukemia cells (RBL cell ghosts) and permeabilized with alpha-toxin from S. aureus are a simplified system for the study of Fc epsilon RI-mediated activation of phospholipase C (PLC). This activity is dependent upon ATP and magnesium, and is enhanced by the addition of another compound containing an energetic phosphate group, either phosphoenolpyruvate (PEP) or phosphocreatine (PCr). This effect appears to be specific for PEP and PCr in that other compounds with energetic phosphate bonds including fructose 1,6-bisphosphate and additional ATP are not effective. On the contrary, GTP-gamma-S, an activator of G proteins, activates PLC in the presence of ATP alone and this is not further enhanced by the addition of PEP. In addition to Fc epsilon RI and GTP-gamma-S, two other stimuli lead to enhanced activity of PLC in permeabilized RBL cell ghosts: 1) an inhibitor of tyrosine phosphatases (Na3VO4) and 2) an analog of adenosine (NECA). Data presented here extend previous results to show that activation of PLC by GTP-gamma-S is not enhanced either by the addition of PCr or by the addition of a more MgATP. Further new findings include the observations that activation of PLC by Na3VO4 is augmented by PEP and PCr in a fashion similar to that observed for Fc epsilon RI-mediated activation of PLC and that activation of PLC by NECA shows even more marked dependency on PEP than does activation by Fc epsilon RI or Na3VO4.(ABSTRACT TRUNCATED AT 250 WORDS)
Dreskin
S C
SC
Department of Medicine, University of Colorado Health Sciences Center, Denver 80262, USA.
D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism* / Q000494:pharmacology; D019830:Adenosine-5'-(N-ethylcarboxamide); D000818:Animals; D000941:Antigens / Q000008:administration & dosage; D001491:Basophils / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D004734:Energy Metabolism; D004789:Enzyme Activation / Q000187:drug effects; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D015471:Leukemia, Basophilic, Acute / Q000201:enzymology; D010716:Phosphatidylinositols / Q000378:metabolism; D010725:Phosphocreatine / Q000494:pharmacology; D010728:Phosphoenolpyruvate / Q000494:pharmacology; D051381:Rats; D017455:Receptors, IgE / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism*; D014638:Vanadates / Q000494:pharmacology
D000941:Antigens; D010716:Phosphatidylinositols; D017455:Receptors, IgE; D010725:Phosphocreatine; D019830:Adenosine-5'-(N-ethylcarboxamide); D016244:Guanosine 5'-O-(3-Thiotriphosphate); D014638:Vanadates; D010728:Phosphoenolpyruvate; D000255:Adenosine Triphosphate; D010738:Type C Phospholipases; D000241:Adenosine