The multifunctional protein GC1q-R interacts specifically with the i3 loop arginine cluster of the vasopressin V2 receptor |
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Authors: | Granier Sébastien Jean-Alphonse Frédéric Bacqueville Daniel Monteil Arnaud Pascal Robert Poncet Joël Guillon Gilles Boudier Laure Arcemisbéhère Laure Mouillac Bernard Bellot Gaétan Déméné Hélène Mendre Christiane |
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Affiliation: | CNRS, UMR 5203, Institut de Génomique fonctionnelle, Montpellier, France. |
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Abstract: | In this study, we identified the multifunctional protein GC1q-R as a novel vasopressin V(2) receptor (V(2)R) interacting protein. For this purpose, we have developed a proteomic approach combining pull-down assays using a cyclic peptide mimicking the third intracellular loop of V(2)R as a bait and mass spectrometry analyses of proteins isolated from either rat or human kidney tissues or the HEK 293 cell line. Co-immunoprecipitation of GC1q-R with the c-Myc-tagged h-V(2)R expressed in a HEK cell line confirmed the existence of a specific interaction between GC1q-R and the V(2) receptor. Then, construction of a mutant receptor in i3 loop allowed us to identify the i3 loop arginine cluster of the vasopressin V(2) receptor as the interacting determinant for GC1q-R interaction. Using purified receptor as a bait and recombinant (74-282) GC1q-R, we demonstrated a direct and specific interaction between these two proteins via the arginine cluster. |
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