Document Detail

Sphingosine-1-phosphate activates phospholipase D in human airway epithelial cells via a G protein-coupled receptor.
MedLine Citation:
PMID:  10683250     Owner:  NLM     Status:  MEDLINE    
Sphingosine-1-phosphate (SPP) acts as a first messenger in immortalized human airway epithelial cells (CFNPE9o(-)), possibly interacting with an Edg family receptor. Expression of the SPP receptors Edg-1 and Edg-3, as well as a low level of Edg-5/H218, was detected in these cells, in agreement with their ability to specifically bind SPP. The related lipids, lysophosphatidic acid and sphingosylphosphorylcholine, were unable to displace SPP from its high affinity binding sites, suggesting that the biological responses to these different lysolipids are mediated by distinct receptors. SPP markedly inhibited forskolin-stimulated cAMP accumulation in a dose-dependent manner and caused a remarkable elevation of intracellular calcium, both effects being sensitive to pertussis toxin treatment. Most importantly, SPP stimulated phosphatidic acid formation, which was maximal after 2 min and decreased within 8-10 min. In the presence of butan-1-ol, suppression of SPP-induced phosphatidic acid formation and production of phosphatidylbutanol were found, clearly indicating activation of phospholipase D (PLD). This finding was also confirmed by analysis of the fatty acid composition of phosphatidic acid, showing an increase in the monounsaturated oleic acid only. The decrease of phosphatidic acid level after 8-10 min incubation with SPP was accompanied by a parallel increase of diacylglycerol production, which was abolished in the presence of butan-1-ol. This result indicates that activation of phospholipase D is followed by stimulation of phosphatidate phosphohydrolase activity. Phosphatidic acid formation was insensitive to protein kinase C inhibitors and almost completely inhibited by pertussis toxin treatment, suggesting that SPP activates phospholipase D via a G(i/o) protein-coupled receptor.
S Orlati; A M Porcelli; S Hrelia; J R Van Brocklyn; S Spiegel; M Rugolo
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Archives of biochemistry and biophysics     Volume:  375     ISSN:  0003-9861     ISO Abbreviation:  Arch. Biochem. Biophys.     Publication Date:  2000 Mar 
Date Detail:
Created Date:  2000-03-24     Completed Date:  2000-03-24     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0372430     Medline TA:  Arch Biochem Biophys     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  69-77     Citation Subset:  IM    
Copyright Information:
Copyright 2000 Academic Press.
Dipart. di Biologia Ev.Sp., Università di Bologna, Bologna, Italy.
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MeSH Terms
3T3 Cells
Binding, Competitive / drug effects
Calcium / metabolism
Cell Line, Transformed
Cyclic AMP / metabolism
DNA-Binding Proteins / genetics,  metabolism
Diglycerides / biosynthesis
Enzyme Activation / drug effects
Enzyme Inhibitors / pharmacology
Epithelial Cells / cytology,  enzymology*
Forskolin / pharmacology
GTP-Binding Proteins / metabolism
I-kappa B Proteins*
Immediate-Early Proteins / genetics,  metabolism
Pertussis Toxin
Phosphatidic Acids / biosynthesis
Phospholipase D / metabolism*
RNA, Messenger / biosynthesis
Receptors, Cell Surface / metabolism*
Receptors, G-Protein-Coupled*
Receptors, Lysophospholipid
Respiratory System / cytology,  enzymology*
Reverse Transcriptase Polymerase Chain Reaction
Signal Transduction / drug effects,  physiology
Sphingosine / analogs & derivatives*,  metabolism,  pharmacology
Virulence Factors, Bordetella / pharmacology
Grant Support
Reg. No./Substance:
0/DNA-Binding Proteins; 0/Diglycerides; 0/Enzyme Inhibitors; 0/I-kappa B Proteins; 0/Immediate-Early Proteins; 0/Lysophospholipids; 0/Phosphatidic Acids; 0/RNA, Messenger; 0/Receptors, Cell Surface; 0/Receptors, G-Protein-Coupled; 0/Receptors, Lysophospholipid; 0/Virulence Factors, Bordetella; 123-78-4/Sphingosine; 139874-52-5/NF-kappaB inhibitor alpha; 26993-30-6/sphingosine 1-phosphate; 60-92-4/Cyclic AMP; 66428-89-5/Forskolin; 7440-70-2/Calcium; EC Toxin; EC D; EC 3.6.1.-/GTP-Binding Proteins

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