Premium
S1P 1 Receptor Regulation by Phosphorylation
Author(s) -
BazúaValenti Silvana,
MorquechoLeón Marco. A,
RomeroÁvila M. Teresa,
GarcíaSáinz J. Adolfo
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.1040.2
Subject(s) - g protein coupled receptor , microbiology and biotechnology , protein kinase c , phosphorylation , internalization , receptor , sphingosine 1 phosphate , sphingosine , biology , lysophosphatidic acid , gene isoform , signal transduction , chemistry , biochemistry , gene
Sphingosine‐1‐phosphate (S1P) is a bioactive molecule that has been implicated in regulating cellular processes. S1P exerts its extracellular effects as a ligand of a GPCR family. S1P 1 receptors are coupled to heterotrimeric G i/o proteins. The importance of S1P and its receptor S1P 1 in multiple cellular and physiological processes (vascular tone, permeability, angiogenesis, lymphocyte egress) makes them and their regulation, an interesting pharmacological target. It has been demonstrated that S1P 1 activation by its natural ligand and the activation of PKC by phorbol myristate acetate leads to receptor phosphorylation and internalization. However, little is known about the identity of the kinases involved in this process. The primary objective of this study is to characterize the molecular processes that lead to receptor phosphorylation and identify the PKC isoform/s implicated. A human prostate cancer cell line stably expressing the S1P 1 fused to the Green Fluorescence Protein was used for this work. We conducted functional analysis by measuring intracellular calcium mobilization in the presence or absence of PMA and distinct PKC selective inhibitors for the different isoforms. S1P 1 ‐PKC interaction in the course of stimulation with PMA was analyzed by co‐immunoprecipitation. Our results indicate that of two isoforms participate in the PMA‐mediated desensitization, PKC α and to a lesser extent PKC β.