Phospholipase C and phospholipase D are activated independently of each other in chemotactic peptide-stimulated human neutrophils

J Leukoc Biol. 1993 Jun;53(6):630-5. doi: 10.1002/jlb.53.6.630.


When cytochalasin B-treated neutrophils were stimulated with fMet-Leu-Phe (fMLP) in the presence of Ca2+, phospholipase C (PLC) activity, as measured by inositol-1,4,5-triphosphate (IP3) formation, preceded phospholipase D (PLD)-catalyzed breakdown of choline-containing phosphoglycerides to form choline and diradyl-sn-glycero-3-phosphate (phosphatidic acid), suggesting a possible link between PLC and PLD. However, in the absence of cytochalasin B or extracellular Ca2+, PLC was fully activated by fMLP with minimal activation of PLD, indicating that PLC activation alone is not sufficient for PLD activation. Full activation of PLD by fMLP required the simultaneous presence of both Ca2+ and cytochalasin B, a condition that caused no further enhancement of PLC. This result suggests that PLD products are not involved in the regulation of PLC activation. Furthermore, under conditions of complete inhibition of PLC by phorbol 12-myristate 13-acetate (PMA), there was no inhibition of PLD, showing that fMLP can activate PLD in the absence of PLC. Treatment of intact neutrophils with pertussis toxin inhibited both PLC and PLD, with PLC inhibition occurring at lower concentrations that PLD inhibition. These differential effects of pertussis toxin and the observed lack of inhibition of fMLP-stimulated PLD by PMA, which is believed to inactivate G-proteins involved in PLC activation, imply that PLC and PLD are linked to fMLP receptors through distinct G-proteins. Taken together, these observations suggest that, in fMLP-stimulated neutrophils, PLC and PLD are activated through independent mechanisms.

MeSH terms

  • Cytochalasin B / pharmacology
  • Enzyme Activation
  • GTP-Binding Proteins / physiology
  • Humans
  • N-Formylmethionine Leucyl-Phenylalanine / pharmacology*
  • Neutrophils / drug effects
  • Neutrophils / enzymology*
  • Pertussis Toxin
  • Phospholipase D / metabolism*
  • Tetradecanoylphorbol Acetate / pharmacology
  • Type C Phospholipases / metabolism*
  • Virulence Factors, Bordetella / pharmacology


  • Virulence Factors, Bordetella
  • Cytochalasin B
  • N-Formylmethionine Leucyl-Phenylalanine
  • Pertussis Toxin
  • Type C Phospholipases
  • Phospholipase D
  • GTP-Binding Proteins
  • Tetradecanoylphorbol Acetate