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J Biol Chem, Vol. 274, Issue 27, 18997-19002, July 2, 1999
From the Center for Vascular Biology, Department of Physiology,
University of Connecticut Health Center, Farmington, Connecticut
06030-3505
Sphingosine 1-phosphate (SPP) is a potent lipid
mediator released upon cellular activation. In this report,
pharmacological properties of the three G-protein-coupled receptors
(GPCRs) for SPP, EDG-1, -3, and -5 are characterized using a
Xenopus oocyte expression system, which lacks endogenous
SPP receptors. Microinjection of the EDG-3 and EDG-5 but not EDG-1
mRNA conferred SPP-responsive intracellular calcium transients;
however, the EDG-5 response was quantitatively much less. Co-expression
of EDG-1 receptor with the chimeric G
qi protein
conferred SPP responsiveness. Gqi or Gq
co-injection also potentiated the EDG-5 and EDG-3 mediated responses to
SPP. These data suggest that SPP receptors couple differentially to the
Gq and Gi pathway. All three GPCRs were also
activated by sphingosylphosphorylcholine, albeit at higher
concentrations. None of the other related sphingolipids tested
stimulated or blocked SPP-induced calcium responses. However, suramin,
a polycyclic anionic compound, selectively antagonized SPP-activated
calcium transients in EDG-3 expressing oocytes with an IC50
of 22 µM, suggesting that it is an antagonist selective for the EDG-3 GPCR isotype. We conclude that the three SPP receptors signal differentially by coupling to different G-proteins. Furthermore, because only EDG-3 was antagonized by suramin, variations in receptor structure may determine differences in antagonist selectivity. This
property may be exploited to synthesize receptor subtype-specific antagonists.
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R. L. Malek, R. E. Toman, L. C. Edsall, S. Wong, J. Chiu, C. A. Letterle, J. R. Van Brocklyn, S. Milstien, S. Spiegel, and N. H. Lee Nrg-1 Belongs to the Endothelial Differentiation Gene Family of G Protein-coupled Sphingosine-1-phosphate Receptors J. Biol. Chem., February 16, 2001; 276(8): 5692 - 5699. [Abstract] [Full Text] [PDF]
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T. Ohmori, Y. Yatomi, H. Okamoto, Y. Miura, G. Rile, K. Satoh, and Y. Ozaki Gi-mediated Cas Tyrosine Phosphorylation in Vascular Endothelial Cells Stimulated with Sphingosine 1-Phosphate. POSSIBLE INVOLVEMENT IN CELL MOTILITY ENHANCEMENT IN COOPERATION WITH Rho-MEDIATED PATHWAYS J. Biol. Chem., February 9, 2001; 276(7): 5274 - 5280. [Abstract] [Full Text] [PDF]
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I. Ishii, B. Friedman, X. Ye, S. Kawamura, C. McGiffert, J. J. A. Contos, M. A. Kingsbury, G. Zhang, J. H. Brown, and J. Chun Selective Loss of Sphingosine 1-Phosphate Signaling with No Obvious Phenotypic Abnormality in Mice Lacking Its G Protein-coupled Receptor, LPB3/EDG-3 J. Biol. Chem., August 31, 2001; 276(36): 33697 - 33704. [Abstract] [Full Text] [PDF]
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