Detection of the human GPR50 orphan seven transmembrane protein by polyclonal antibodies mapping different epitopes

Autor: Hassina Ould Hamouda, Pascal Maurice, Ralf Jockers, Nejla Sözer-Topçular, Jean-Luc Guillaume, Avais M. Daulat, Gilles Ferry, Jean A. Boutin, Angélique Levoye, Rivka Ravid, Patty Chen, Egemen Savaskan, Philippe Delagrange
Přispěvatelé: Institut Cochin (UMR_S567 / UMR 8104), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), University of Basel (Unibas), Netherlands Brain Bank [Amsterdam, The Netherlands] (N2B), Institut de Recherches Internationales Servier [Suresnes] (IRIS), Centre de Recherche en Cancérologie de Marseille (CRCM), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Aix Marseille Université (AMU), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de Recherches Servier, Centre de Recherches de Croissy, Département des Sciences Expérimentales, Institut de Recherches Internationales Servier, University of Zurich, Jockers, R, MAURICE, Pascal
Jazyk: angličtina
Rok vydání: 2007
Předmět:
[SDV]Life Sciences [q-bio]
Molecular Sequence Data
Nerve Tissue Proteins
610 Medicine & health
CHO Cells
Immunofluorescence
Antibodies
Epitope
Cell Line
Receptors
G-Protein-Coupled
Epitopes
03 medical and health sciences
Cricetulus
0302 clinical medicine
Endocrinology
Western blot
Sequence Analysis
Protein
Cricetinae
medicine
Animals
Humans
Amino Acid Sequence
ComputingMilieux_MISCELLANEOUS
030304 developmental biology
Orphan receptor
0303 health sciences
biology
medicine.diagnostic_test
11359 Institute for Regenerative Medicine (IREM)
Fusion protein
Molecular biology
Transmembrane protein
Protein Structure
Tertiary
1310 Endocrinology
[SDV] Life Sciences [q-bio]
Epitope mapping
Polyclonal antibodies
biology.protein
Binding Sites
Antibody
Rabbits
Epitope Mapping
030217 neurology & neurosurgery
Zdroj: Journal of Pineal Research
Journal of Pineal Research, Wiley, 2007, 43 (1), pp.10-15. ⟨10.1111/j.1600-079X.2007.00437.x⟩
Journal of Pineal Research, 2007, 43 (1), pp.10-15. ⟨10.1111/j.1600-079X.2007.00437.x⟩
ISSN: 0742-3098
1600-079X
DOI: 10.1111/j.1600-079X.2007.00437.x⟩
Popis: International audience; GPR50 is an orphan seven transmembrane protein related to the melatonin receptor subfamily comprising MT(1) and MT(2) receptors. In the absence of any known ligand for GPR50, other tools are critical for the characterization of this protein. Here, we describe the generation, purification and characterization of the first rabbit polyclonal antibodies generated against peptides corresponding to the N-terminus, C-terminus and two additional regions within the intracellular tail of GPR50. Immune sera were purified on peptide-antigen affinity columns. Antibodies specifically recognized a GPR50-YFP fusion protein on the plasma membrane of HEK 293 cells in immunofluorescence experiments. In Western blot experiments, the monomeric and dimeric forms of GPR50 were detected as proteins of 66 and 130 kDa, respectively. In addition, these new antibodies were sufficiently sensitive to detect GPR50 in brain slices of the rat pituitary and human hippocampus. In conclusion, we successfully produced antibodies against the orphan GPR50 protein that will become valuable tools for functional studies of this protein.
Databáze: OpenAIRE
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