Apparent loss-of-function mutant GPCRs revealed as constitutively desensitized receptors

被引:68
作者
Wilbanks, AM
Laporte, SA
Bohn, LM
Barak, LS
Caron, MG
机构
[1] Duke Univ, Med Ctr, Howard Hughes Med Inst Labs, Durham, NC 27710 USA
[2] Duke Univ, Med Ctr, Dept Cell Biol, Durham, NC 27710 USA
[3] Duke Univ, Med Ctr, Dept Med, Durham, NC 27710 USA
关键词
D O I
10.1021/bi020275m
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The DRY motif is a triplet amino acid sequence (aspartic acid, arginine, and tyrosine) that is highly conserved in G protein-coupled receptors (GPCRs). Recently, we have shown that a molecular determinant for nephrogenic diabetes insipidus, the vasopressin receptor with a substitution at the DRY motif arginine (V2R R137H), is a constitutively desensitized receptor that is unable to couple to G proteins due to its constitutive association with beta-arrestin [Barak, L. S. (2001) Proc. Natl. Acad. Sci. U.S.A. 98, 93-98]. Additionally, the mutant receptors are localized in endocytic vesicles, identical to wild-type receptors stimulated with agonist. In this study, we asked whether the constitutively desensitized phenotype observed in the V2R R137H represents a general paradigm that may be extended to other GPCRs. We show that arginine substitutions in the DRY motifs of the alpha(1B) adrenergic receptor (alpha(1B)-AR) and angiotensin II type IA receptor (AT(1A)R) result in receptors that are uncoupled from G proteins, associated with beta-arrestins, and found localized in endocytic vesicles rather than at the plasma membrane in the absence of agonists. The localization of the alpha(1B)-ARs and AT(1A)Rs with arginine substitutions can be restored,to the plasma membrane by either using selective antagonists or preventing the endocytosis of the beta-arrestin-receptor complexes. These results indicate that the arginine residue of the DRY motif is essential for preserving the localization of the inactive receptor complex. Furthermore, constitutive desensitization may underlie some loss-of-function receptor phenotypes and represent an unappreciated mechanism of hormonal resistance.
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页码:11981 / 11989
页数:9
相关论文
共 27 条
[1]   Receptor/β-arrestin complex formation and the differential trafficking and resensitization of β2-adrenergic and angiotensin II type 1A receptors [J].
Anborgh, PH ;
Seachrist, JL ;
Dale, LB ;
Ferguson, SSG .
MOLECULAR ENDOCRINOLOGY, 2000, 14 (12) :2040-2053
[2]   THE CONSERVED 7-TRANSMEMBRANE SEQUENCE NP(X)(2,3)Y OF THE G-PROTEIN-COUPLED RECEPTOR SUPERFAMILY REGULATES MULTIPLE PROPERTIES OF THE BETA(2)-ADRENERGIC RECEPTOR [J].
BARAK, LS ;
MENARD, L ;
FERGUSON, SSG ;
COLAPIETRO, AM ;
CARON, MG .
BIOCHEMISTRY, 1995, 34 (47) :15407-15414
[3]   A beta-arrestin green fluorescent protein biosensor for detecting G protein-coupled receptor activation [J].
Barak, LS ;
Ferguson, SSG ;
Zhang, J ;
Caron, MG .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (44) :27497-27500
[4]   Constitutive arrestin-mediated desensitization of a human vasopressin receptor mutant associated with nephrogenic diabetes insipidus [J].
Barak, LS ;
Oakley, RH ;
Laporte, SA ;
Caron, MG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (01) :93-98
[5]   MUTATIONS AND DISEASES OF G-PROTEIN COUPLED RECEPTORS [J].
BIRNBAUMER, M .
JOURNAL OF RECEPTOR AND SIGNAL TRANSDUCTION RESEARCH, 1995, 15 (1-4) :131-160
[6]  
CICCARONE V, 1999, FOCUS, V21, P54
[7]   SIGNALING AND REGULATION OF THE ALPHA(1B)-ADRENERGIC RECEPTOR [J].
COTECCHIA, S ;
LATTION, AL ;
DIVIANI, D ;
CAVALLI, A .
BIOCHEMICAL SOCIETY TRANSACTIONS, 1995, 23 (01) :121-125
[8]  
COTECCHIA S, 1992, J BIOL CHEM, V267, P1633
[9]  
Ferguson SSG, 2001, PHARMACOL REV, V53, P1
[10]   beta-arrestin acts as a clathrin adaptor in endocytosis of the beta(2)-adrenergic receptor [J].
Goodman, OB ;
Krupnick, JG ;
Santini, F ;
Gurevich, VV ;
Penn, RB ;
Gagnon, AW ;
Keen, JH ;
Benovic, JL .
NATURE, 1996, 383 (6599) :447-450