Sphingosylphosphorylcholine and Lysophosphatidylcholine Are Ligands for the G Protein-coupled Receptor GPR4

Kui Zhu, Linnea M. Baudhuin, Guiying Hong, Freager S. Williams, Kelly L. Cristina, Janusz H S Kabarowski, Owen N. Wittell, Yan Xu

Research output: Contribution to journalArticle

197 Citations (Scopus)

Abstract

Sphingosylphosphorylcholine (SPC) and lysophosphatidylcholine (LPC) are bioactive lipid molecules involved in numerous biological processes. We have recently identified ovarian cancer G protein-coupled receptor 1 (OGR1) as a specific and high affinity receptor for SPC, and G2A as a receptor with high affinity for LPC, but low affinity for SPC. Among G protein-coupled receptors, GPR4 shares highest sequence homology with OGR1 (51%). In this work, we have identified GPR4 as not only another high affinity receptor for SPC, but also a receptor for LPC, albeit of lower affinity. Both SPC and LPC induce increases in intracellular calcium concentration in GPR4-, but not vector-transfected MCF10A cells. These effects are insensitive to treatment with BN52021, WEB-2170, and WEB-2086 (specific platelet activating factor (PAF) receptor antagonists), suggesting that they are not mediated through an endogenous PAF receptor. SPC and LPC bind to GPR4 in GPR4-transfected CHO cells with Kd/SPC = 36 nM, and Kd/LPC = 159 nM, respectively. Competitive binding is elicited only by SPC and LPC. Both SPC and LPC activate GPR4-dependent activation of serum response element reporter and receptor internalization. Swiss 3T3 cells expressing GPR4 respond to both SPC and LPC, but not sphingosine 1-phosphate (S1P), PAF, psychosine (Psy), glucosyl-β1′ 1-sphingosine (Glu-Sph), galactosyl-β1′1-ceramide (GalCer), or lactosyl-β1′1-ceramide (Lac-Cer) to activate extracellular signal-regulated kinase mitogen-activated protein kinase in a concentration- and time-dependent manner. SPC and LPC stimulate DNA synthesis in GPR4-expressing Swiss 3T3 cells. Both extracellular signal-regulated kinase activation and DNA synthesis stimulated by SPC and LPC are pertussis toxin-sensitive, suggesting the involvement of a Gi-heterotrimeric G protein. In addition, GPR4 expression confers chemotactic responses to both SPC and LPC in Swiss 3T3 cells. Taken together, our data indicate that GPR4 is a receptor with high affinity to SPC and low affinity to LPC, and that multiple cellular functions can be transduced via this receptor.

Original languageEnglish (US)
Pages (from-to)41325-41335
Number of pages11
JournalJournal of Biological Chemistry
Volume276
Issue number44
DOIs
StatePublished - Nov 2 2001
Externally publishedYes

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Lysophosphatidylcholines
G-Protein-Coupled Receptors
Ligands
Swiss 3T3 Cells
Ceramides
Extracellular Signal-Regulated MAP Kinases
WEB 2086
sphingosine phosphorylcholine
Lactosylceramides
Serum Response Element
Chemical activation
Galactosylceramides
Biological Phenomena
Heterotrimeric GTP-Binding Proteins
Sphingosine
Competitive Binding
CHO Cells
Platelet Activating Factor
DNA
Pertussis Toxin

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sphingosylphosphorylcholine and Lysophosphatidylcholine Are Ligands for the G Protein-coupled Receptor GPR4. / Zhu, Kui; Baudhuin, Linnea M.; Hong, Guiying; Williams, Freager S.; Cristina, Kelly L.; Kabarowski, Janusz H S; Wittell, Owen N.; Xu, Yan.

In: Journal of Biological Chemistry, Vol. 276, No. 44, 02.11.2001, p. 41325-41335.

Research output: Contribution to journalArticle

Zhu, K, Baudhuin, LM, Hong, G, Williams, FS, Cristina, KL, Kabarowski, JHS, Wittell, ON & Xu, Y 2001, 'Sphingosylphosphorylcholine and Lysophosphatidylcholine Are Ligands for the G Protein-coupled Receptor GPR4', Journal of Biological Chemistry, vol. 276, no. 44, pp. 41325-41335. https://doi.org/10.1074/jbc.M008057200
Zhu, Kui ; Baudhuin, Linnea M. ; Hong, Guiying ; Williams, Freager S. ; Cristina, Kelly L. ; Kabarowski, Janusz H S ; Wittell, Owen N. ; Xu, Yan. / Sphingosylphosphorylcholine and Lysophosphatidylcholine Are Ligands for the G Protein-coupled Receptor GPR4. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 44. pp. 41325-41335.
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AU - Williams, Freager S.

AU - Cristina, Kelly L.

AU - Kabarowski, Janusz H S

AU - Wittell, Owen N.

AU - Xu, Yan

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N2 - Sphingosylphosphorylcholine (SPC) and lysophosphatidylcholine (LPC) are bioactive lipid molecules involved in numerous biological processes. We have recently identified ovarian cancer G protein-coupled receptor 1 (OGR1) as a specific and high affinity receptor for SPC, and G2A as a receptor with high affinity for LPC, but low affinity for SPC. Among G protein-coupled receptors, GPR4 shares highest sequence homology with OGR1 (51%). In this work, we have identified GPR4 as not only another high affinity receptor for SPC, but also a receptor for LPC, albeit of lower affinity. Both SPC and LPC induce increases in intracellular calcium concentration in GPR4-, but not vector-transfected MCF10A cells. These effects are insensitive to treatment with BN52021, WEB-2170, and WEB-2086 (specific platelet activating factor (PAF) receptor antagonists), suggesting that they are not mediated through an endogenous PAF receptor. SPC and LPC bind to GPR4 in GPR4-transfected CHO cells with Kd/SPC = 36 nM, and Kd/LPC = 159 nM, respectively. Competitive binding is elicited only by SPC and LPC. Both SPC and LPC activate GPR4-dependent activation of serum response element reporter and receptor internalization. Swiss 3T3 cells expressing GPR4 respond to both SPC and LPC, but not sphingosine 1-phosphate (S1P), PAF, psychosine (Psy), glucosyl-β1′ 1-sphingosine (Glu-Sph), galactosyl-β1′1-ceramide (GalCer), or lactosyl-β1′1-ceramide (Lac-Cer) to activate extracellular signal-regulated kinase mitogen-activated protein kinase in a concentration- and time-dependent manner. SPC and LPC stimulate DNA synthesis in GPR4-expressing Swiss 3T3 cells. Both extracellular signal-regulated kinase activation and DNA synthesis stimulated by SPC and LPC are pertussis toxin-sensitive, suggesting the involvement of a Gi-heterotrimeric G protein. In addition, GPR4 expression confers chemotactic responses to both SPC and LPC in Swiss 3T3 cells. Taken together, our data indicate that GPR4 is a receptor with high affinity to SPC and low affinity to LPC, and that multiple cellular functions can be transduced via this receptor.

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