Document Detail

Phosphatidylcholine-specific phospholipase D-derived 1,2-diacylglycerol does not initiate protein kinase C activation in the RBL 2H3 mast-cell line.
MedLine Citation:
PMID:  1384468     Owner:  NLM     Status:  MEDLINE    
We examined the role of phosphatidylcholine-specific phospholipase D (PC-PLD) in the IgE-dependent activation of protein kinase C (PKC) in RBL 2H3 cells (a model for mast-cell function). Cells were sensitized with mouse monoclonal anti-trinitrophenol (TNP) IgE (0.5 micrograms/ml) and were then triggered with an optimal concentration (10 ng/ml) of TNP-ovalbumin conjugate (TNP-OVA). This resulted in an immediate biphasic increase in the production of 1,2-diacylglycerol (DAG) and activation of PKC. The initial increase in DAG production reached a peak within 30 s, and the second phase reached a plateau within 5 min after stimulation. TNP-OVA-induced PC-PLD activation followed the initial increase in DAG formation in response to IgE-receptor cross-bridging, but coincided with the second peak. Phosphatidic acid (PA), derived from the PC-PLD pathway, is metabolized to DAG by the action of PA phosphohydrolase (PAPase). Propranolol (0.3 mM), which inhibits PAPase, blocked the IgE-dependent increase in DAG, activation of PKC, and subsequently degranulation. The PKC inhibitor staurosporine (0.1 microM) inhibited the second, but not first, peak of DAG accumulation, reversed PKC translocation after 10 min and inhibited subsequent mediator release. Taken together, these results demonstrate that PC-PLD does not initiate, but may play a latent role in, IgE-dependent DAG production, PKC activation and mediator release from RBL 2H3 cells.
P Lin; W J Fung; A M Gilfillan
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Publication Detail:
Type:  In Vitro; Journal Article    
Journal Detail:
Title:  The Biochemical journal     Volume:  287 ( Pt 1)     ISSN:  0264-6021     ISO Abbreviation:  Biochem. J.     Publication Date:  1992 Oct 
Date Detail:
Created Date:  1992-11-10     Completed Date:  1992-11-10     Revised Date:  2010-09-07    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  325-31     Citation Subset:  IM    
Department of Pharmacology, Hoffmann-La Roche, Nutley, NJ 07110.
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MeSH Terms
Alkaloids / pharmacology
Cell Compartmentation / drug effects
Diglycerides / metabolism*
Enzyme Activation / drug effects
Ethanol / pharmacology
Histamine Release / drug effects
Mast Cells / metabolism*
Phosphatidylcholines / metabolism*
Phospholipase D / metabolism*
Propranolol / pharmacology
Protein Kinase C / antagonists & inhibitors,  metabolism*
Receptors, IgE / physiology
Signal Transduction
Tumor Cells, Cultured
Reg. No./Substance:
0/Alkaloids; 0/Diglycerides; 0/Phosphatidylcholines; 0/Receptors, IgE; 525-66-6/Propranolol; 62996-74-1/Staurosporine; 64-17-5/Ethanol; EC Kinase C; EC D

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