Lysophosphatidic acid is a mediator of Trp-Lys-Tyr-Met-Val-d-Met-induced calcium influx

doi:10.1016/j.bbrc.2004.09.072

Biochemical and Biophysical Research Communications

Volume 324, Issue 1, 5 November 2004, Pages 458-465

https://doi.org/10.1016/j.bbrc.2004.09.072 Get rights and content

Abstract

Intracellular calcium (Ca²⁺) homeostasis is very strictly regulated, and the activation of G-protein-coupled receptor (GPCR) can cause two different calcium changes, intracellular calcium release, and calcium influx. In this study, we investigated the possible role of lysophosphatidic acid (LPA) on GPCR-induced Ca²⁺ signaling. The addition of exogenous LPA induced dramatic Ca²⁺ influx but not intracellular Ca²⁺ release in U937 cells. LPA-induced Ca²⁺ influx was not affected by pertussis toxin and phospholipase C inhibitor (U73122), ruling out the involvement of pertussis toxin-sensitive G-proteins, and phospholipase C. Stimulation of U937 cells with Trp-Lys-Tyr-Met-Val-d-Met (WKYMVm), which binds to formyl peptide receptor like 1, enhanced phospholipase A₂ and phospholipase D activation, indicating LPA formation. The inhibition of LPA synthesis by phospholipase A₂-specific inhibitor (MAFP) or n-butanol significantly inhibited WKYMVm-induced Ca²⁺ influx, suggesting a crucial role for LPA in the process. Taken together, we suggest that LPA mediates WKYMVm-induced Ca²⁺ influx.

Section snippets

Materials and methods

Materials. Lysophosphatidic acid (1-myristyl-2-hydroxy-sn-glycero-3-phosphate, sodium salt), lysophosphatidylcholine (1-myristyl-2-hydroxy-sn-glycero-phosphocholine), lysophosphatidylethanolamine (1-myristyl-2-hydroxy-sn-glycero-phosphoethanolamine), and lysophosphatidylserine (1-myristyl-2-hydroxy-sn-glycero-phosphoserine) were from Avanti polar lipids (Alabaster, AL). Sphingosine-1-phosphate, phytosphingosine-1-phosphate, MAFP (methyl arachidonylfluorophosphonate), AACOCF₃

LPA stimulates Ca²⁺ influx without Ca²⁺ release in U937 cells

In this study, we examined the effect of LPA on calcium influx in U937 cells. When U937 cells were stimulated with various concentrations of LPA in the absence of extracellular Ca²⁺, no significant change in the cytosolic Ca²⁺ concentration was observed (Fig. 1A). However, the addition of extracellular Ca²⁺ to LPA-stimulated U937 cells caused a dramatic Ca²⁺ influx (Fig. 1A). This LPA-induced Ca²⁺ influx increased according to the added Ca²⁺ concentration and showed maximal activity at around 1

Discussion

This study demonstrates that LPA plays a role in the regulation of Ca²⁺ influx in U937 cells. Moreover, the generation of LPA is crucially required for GPCR (FPRL1)-induced Ca²⁺ influx in U937 cells. Since many reports have demonstrated the activation of cell surface receptors, such as LPA₁, LPA₂, and LPA₃ by LPA, we also checked whether U937 cells express cell surface LPA receptors by RT-PCR analysis. We found that U937 cells express the mRNA transcript of LPA₂, but not those of LPA₁ and LPA₃

Acknowledgments

This work was supported by Grant 02-PJ1-PG10-20706-0001 from the Korea Health 21 R&D Project, Ministry of Health and Welfare, Republic of Korea (to Y.S.B.). We thank Dr. D.S. Im at Pusan National University College of Pharmacy for his helpful discussion.

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Lysophosphatidic acid is a mediator of Trp-Lys-Tyr-Met-Val-d-Met-induced calcium influx

Abstract

Section snippets

Materials and methods

LPA stimulates Ca2+ influx without Ca2+ release in U937 cells

Discussion

Acknowledgments

Cell Calcium

Biochim. Biophys. Acta

J. Biol. Chem.

Exp. Cell Res.

J. Biol. Chem.

Prostaglandins

J. Biol. Chem.

Blood

Cell. Signal.

Cell

Curr. Biol.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Prog. Lipid Res.

J. Biol. Chem.

Inositol trisphosphate and calcium signaling

Nature

Mechanisms of calcium signaling by cyclic ADP–ribose and NAADP

Physiol. Rev.

LPA stimulates Ca²⁺ influx without Ca²⁺ release in U937 cells