Sphingosylphosphorylcholine and lysophosphatidylcholine are ligands for the G protein-coupled receptor GPR4

J Biol Chem. 2001 Nov 2;276(44):41325-35. doi: 10.1074/jbc.M008057200. Epub 2001 Sep 4.


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 K(d)/SPC = 36 nm, and K(d)/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-beta1'1-sphingosine (Glu-Sph), galactosyl-beta1'1-ceramide (Gal-Cer), or lactosyl-beta1'1-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 G(i)-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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.
  • Retracted Publication

MeSH terms

  • 3T3 Cells
  • Animals
  • Base Sequence
  • CHO Cells
  • Calcium / metabolism
  • Cricetinae
  • DNA Primers
  • DNA Replication
  • Enzyme Activation
  • Humans
  • Ligands
  • Lysophosphatidylcholines / metabolism*
  • Mice
  • Mitogen-Activated Protein Kinases / metabolism
  • Phosphorylcholine / analogs & derivatives*
  • Phosphorylcholine / metabolism*
  • Radioligand Assay
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism*
  • Receptors, G-Protein-Coupled*
  • Sphingosine / analogs & derivatives*
  • Sphingosine / metabolism*
  • Transfection


  • DNA Primers
  • GPR4 protein, human
  • Ligands
  • Lysophosphatidylcholines
  • Receptors, Cell Surface
  • Receptors, G-Protein-Coupled
  • sphingosine phosphorylcholine
  • Phosphorylcholine
  • Mitogen-Activated Protein Kinases
  • Sphingosine
  • Calcium