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Amino acids within intracellular loop 3 of the D2 dopamine receptor are necessary for arrestin binding and receptor internalization
Author(s) -
Clayton Cecilea C,
Neve Kim A
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.1172.7
Subject(s) - internalization , arrestin , dopamine receptor d2 , 5 ht5a receptor , g protein coupled receptor , receptor , microbiology and biotechnology , g protein coupled receptor kinase , enzyme linked receptor , biology , dopaminergic , dopamine receptor , chemistry , signal transduction , dopamine , biochemistry , endocrinology
Arrestins facilitate dopamine D2 receptor internalization and desensitization and are important mediators of dopaminergic behaviors. Using bioluminescence resonance energy transfer (BRET) to measure receptor‐mediated recruitment of arrestin, our objective was to determine the smallest mutation that prevents arrestin binding while retaining signaling through G‐proteins. Mutation of four residues in intracellular loop three (212–215, IYIV) abolished arrestin recruitment and internalization, but impaired receptor expression. Mutation of three (YIV) or two (IV) residues reduced arrestin recruitment by forty and thirty percent respectively, without impairing receptor expression. Importantly, mutating three residues rendered the receptor incapable of internalization, even when over‐expressing arrestin3 and GRK2, while sparing the receptor's ability to signal through G‐proteins. Thus, mutation of the sequence YIV in IC3 of the D2 receptor selectively impaired arrestin recruitment and abolished receptor internalization, suggesting that this mutant will be a useful tool for elucidating cellular and behavioral effects of D2 receptor stimulation that are mediated by arrestins. (VHA, MH045372 and DA007262 )