Characterization of a novel subtype of human G protein-coupled receptor for lysophosphatidic acid

The recent identification of the Vzg-1/Edg2 protein as a functional G protein coupled receptor for lysophosphatidic acid (LPA) has allowed a sequence based search for new ones that may encode novel subtypes of LPA receptors. A human cDNA encoding a G protein-coupled receptor, designated Edg4, was identified by searching the GenBank(TM) for homologs of the human Ed2 LPA receptor. The Edg4 protein is 46% identical and 72% similar in amino acid sequence to human Edg2. When overexpressed in Jurkat T cells, the Edg4 protein mediated LPA-induced activation of a serum response element reporter gene with LPR concentration dependence (EC50 of 10 nM) and specificity, This LPA-induced reporter gene activation could be partially inhibited by pretreatment with petussis toxin or C3 exoenzyme, suggesting requirements for both a G(i) protein and Rho GTPase. Overexpression of Edg4 in Jurkat cells also led to increases in specific binding sites for [H-3]LPA. Northern blots revealed that two edg4 mRNA transcripts of 1.8 and 8 kilobases are distributed very differently from edg2 nRNAs in adult human tissues and several cancer cell lines. The existence and distinctive tissue expression of structurally different subtypes of LPA receptors may provide one basis for tissue-specific functions and permit independent regulation of each subtype of LPA receptor.