Inhibition of formyl-methionyl-leucyl-phenylalanine-stimulated phospholipase D activation in rat neutrophils by the synthetic isoquinoline DMDI
Copyright policy )Inhibition of formyl-methionyl-leucyl-phenylalanine-stimulated phospholipase D activation in rat neutrophils by the synthetic isoquinoline DMDI
The expression of phospholipase D (PLD) isoenzymes in neutrophils was investigated using reverse transcription-polymerase chain reaction analysis. Amplification products of predicted size were obtained from rat neutrophils with nucleotide sequences corresponding to PLD1a and PLD2. 1-(3',4'-Dimethoxybenzyl)-6,7-dichloroisoquinoline (DMDI) inhibited the formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated PLD activation in rat neutrophils. The underlying cellular signaling mechanism of DMDI inhibition was investigated. The fMLP-induced protein tyrosine phosphorylation and the membrane translocation of ADP-ribosylation factor (ARF) and Rho A in neutrophils was attenuated by DMDI in a concentration-dependent manner. However, neither the membrane association of protein kinase C-alpha and -beta isoenzymes in fMLP-stimulated cells nor the GTPgammaS- and phorbol 12-myristate 13-acetate-stimulated membrane translocation of ARF and Rho A in a cell-free system was affected significantly by DMDI. These results indicate that the expression of PLD1a and PLD2 mRNA in neutrophils. Attenuation of protein tyrosine phosphorylation and the membrane association of ARF and Rho A probably play a concerted role in the inhibition of PLD by DMDI in rat neutrophils in response to fMLP.
- Taichung Veterans General Hospital Taiwan
- Taipei Medical University Taiwan
Cell-Free System, Neutrophils, Reverse Transcriptase Polymerase Chain Reaction, Cell Membrane, Isoquinolines, Rats, Enzyme Activation, Isoenzymes, N-Formylmethionine Leucyl-Phenylalanine, Phospholipase D, Animals, ADP-Ribosylation Factor 1, RNA, Messenger, Enzyme Inhibitors, Phosphorylation, rhoA GTP-Binding Protein, Protein Kinase C
Cell-Free System, Neutrophils, Reverse Transcriptase Polymerase Chain Reaction, Cell Membrane, Isoquinolines, Rats, Enzyme Activation, Isoenzymes, N-Formylmethionine Leucyl-Phenylalanine, Phospholipase D, Animals, ADP-Ribosylation Factor 1, RNA, Messenger, Enzyme Inhibitors, Phosphorylation, rhoA GTP-Binding Protein, Protein Kinase C
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