Solís, K.H.; Romero-Ávila, M.T.; Rincón-Heredia, R.; García-Sáinz, J.A. LPA3 Receptor Phosphorylation Sites: Roles in Signaling and Internalization. Preprints2024, 2024040541. https://doi.org/10.20944/preprints202404.0541.v1
APA Style
Solís, K.H., Romero-Ávila, M.T., Rincón-Heredia, R., & García-Sáinz, J.A. (2024). LPA3 Receptor Phosphorylation Sites: Roles in Signaling and Internalization. Preprints. https://doi.org/10.20944/preprints202404.0541.v1
Chicago/Turabian Style
Solís, K.H., Ruth Rincón-Heredia and J. Adolfo García-Sáinz. 2024 "LPA3 Receptor Phosphorylation Sites: Roles in Signaling and Internalization" Preprints. https://doi.org/10.20944/preprints202404.0541.v1
Abstract
Lysophosphatidic acid (LPA) type 3 (LPA3) receptor mutants were generated in which the sites detected phosphorylated were substituted by non-phosphorylatable amino acids. Substitutions were made in the intracellular loop 3 (IL3 mutant), the carboxyl terminus (Ctail), and both domains (IL3/Ctail). The wild-type (WT) receptor and the mutants were expressed in T-REx HEK293 cells, and the consequences of the substitutions were analyzed employing different functional parameters. Agonist- and LPA-mediated receptor phosphorylation was diminished in the IL3 and Ctail mutants and essentially abolished in the IL3/Ctail mutant, indicating that the main sites were substituted and distributed in both domains. The WT and mutant receptors increased intracellular calcium and ERK 1/2 phosphorylation in response to LPA and PMA. An increase in baseline ERK phosphorylation was detected in the IL3/Ctail mutant. LPA and PMA-induced receptor interaction with β-arrestin 2 and LPA3 internalization were severely diminished in the mutants. Mutant-expressing cells also exhibit increased baseline proliferation and response to different stimuli, which were inhibited by the antagonist Ki16425, suggesting a role of LPA receptors in this process. Migration in response to different attractants was markedly increased in the Ctail mutant, which the Ki16425 antagonist also attenuated.
Medicine and Pharmacology, Pharmacology and Toxicology
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