Role of the C terminus in neuropeptide YY1 receptor desensitization and internalization

被引:36
作者
Holliday, ND [1 ]
Lam, CW [1 ]
Tough, IR [1 ]
Cox, HM [1 ]
机构
[1] Kings Coll London, Wolfson Ctr Age Related Dis, London, England
关键词
D O I
10.1124/mol.104.006114
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
We have studied truncation mutants of the rat neuropeptide Y (NPY) Y-1 receptor lacking four (Thr361stop, Y1T361*) or eight (Ser352stop, Y1S352*) potential serine/threonine C-terminal phosphorylation sites. NPY-stimulated hemagglutinin-tagged Y1, Y1T361*, and Y1S352* receptors all efficiently activated G proteins in Chinese hamster ovary (CHO) cell membranes, but desensitization after NPY pretreatment was only prevented in the HAY1S352* clone. In transfected colonic carcinoma epithelial layers, functional Y1 and Y1T361* peptide YY responses became more transient as the agonist concentration increased, whereas those mediated by the Y1S352* receptor remained sustained. NPY-stimulated HAY1 receptor phosphorylation was increased by transient overexpression of G protein-coupled receptor kinase 2, and only Ser352stop truncation abolished this response in CHO or human embryonic kidney (HEK) 293 cells. Rapid internalization of cell-surface HAY1 receptors in HEK293 cells was observed in response to agonist, resulting in partial colocalization with transferrin, a marker for clathrin-mediated endocytosis and recycling. It is surprising that both truncated receptors were constitutively internalized, predominantly in transferrin-positive compartments. NPY increased cell-surface localization of HAY1S352* receptors, whereas the distribution of both mutants was unaltered by BIBO3304. Recruitment of green fluorescent protein-tagged beta-arrestin2 to punctate endosomes was observed only for HAY1 and HAY1T361* receptors and solely under NPY-stimulated conditions. Thus, the key C-terminal sequence between Ser352 and Lys360 is a major site for Y1 receptor phosphorylation, is critical for its desensitization, and contributes to the association between the receptor and beta-arrestin proteins. However, additional beta-arrestin-independent mechanisms control Y1 receptor trafficking under basal conditions.
引用
收藏
页码:655 / 664
页数:10
相关论文
共 40 条
[1]   The basolateral sorting signals of the thyrotropin and luteinizing hormone receptors:: An unusual family of signals sharing an unusual distal intracellular localization, but unrelated in their structures [J].
Beau, I ;
Groyer-Picard, MT ;
Desroches, A ;
Condamine, E ;
Leprince, J ;
Tomé, JP ;
Dessen, P ;
Vaudry, H ;
Misrahi, M .
MOLECULAR ENDOCRINOLOGY, 2004, 18 (03) :733-746
[2]   The use of bioluminescence resonance energy transfer 2 to study neuropeptide Y receptor agonist-induced β-arrestin 2 interaction [J].
Berglund, MM ;
Schober, DA ;
Statnick, MA ;
McDonald, PH ;
Gehlert, DR .
JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS, 2003, 306 (01) :147-156
[3]   REMOVAL OF PHOSPHORYLATION SITES FROM THE BETA-2-ADRENERGIC RECEPTOR DELAYS ONSET OF AGONIST-PROMOTED DESENSITIZATION [J].
BOUVIER, M ;
HAUSDORFF, WP ;
DEBLASI, A ;
ODOWD, BF ;
KOBILKA, BK ;
CARON, MG ;
LEFKOWITZ, RJ .
NATURE, 1988, 333 (6171) :370-373
[4]   Phosphorylation of key serine residues is required for internalization of the complement 5a (C5a) anaphylatoxin receptor via a β-arrestin, dynamin, and clathrin-dependent pathway [J].
Braun, L ;
Christophe, T ;
Boulay, F .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (06) :4277-4285
[5]   Signaling at zero G: G-protein-independent functions for 7-TM receptors [J].
Brzostowski, JA ;
Kimmel, AR .
TRENDS IN BIOCHEMICAL SCIENCES, 2001, 26 (05) :291-297
[6]   Neuropeptide Y, Y1, Y2 and Y4 receptors mediate Y agonist responses in isolated human colon mucosa [J].
Cox, HM ;
Tough, IR .
BRITISH JOURNAL OF PHARMACOLOGY, 2002, 135 (06) :1505-1512
[7]   Endocytosis of the viral chemokine receptor US28 does not require beta-arrestins but is dependent on the clathrin-mediated pathway [J].
Fraile-Ramos, A ;
Kohout, TA ;
Waldhoer, M ;
Marsh, M .
TRAFFIC, 2003, 4 (04) :243-253
[8]   Neuropeptide Y inhibits capsaicin-sensitive nociceptors via a Y1-receptor-mediated mechanism [J].
Gibbs, J ;
Flores, CM ;
Hargreaves, KM .
NEUROSCIENCE, 2004, 125 (03) :703-709
[9]   Rapid internalization and recycling of the human neuropeptide YY1 receptor [J].
Gicquiaux, H ;
Lecat, S ;
Gaire, M ;
Dieterlen, A ;
Mély, Y ;
Takeda, K ;
Bucher, B ;
Galzi, JL .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (08) :6645-6655
[10]   The molecular acrobatics of arrestin activation [J].
Gurevich, VV ;
Gurevich, EV .
TRENDS IN PHARMACOLOGICAL SCIENCES, 2004, 25 (02) :105-111