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Titolo:
Agonist-promoted trafficking of human bradykinin receptors: arrestin- and dynamin-independent sequestration of the B-2 receptor and bradykinin in HEK293 cells
Autore:
Lamb, ME; De Weerd, WFC; Leeb-Lundberg, LMF;
Indirizzi:
Univ Texas, Hlth Sci Ctr, Dept Biochem, San Antonio, TX 78229 USA Univ Texas San Antonio TX USA 78229 pt Biochem, San Antonio, TX 78229 USA
Titolo Testata:
BIOCHEMICAL JOURNAL
, volume: 355, anno: 2001,
parte:, 3
pagine: 741 - 750
SICI:
0264-6021(20010501)355:<741:ATOHBR>2.0.ZU;2-B
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROTEIN-COUPLED RECEPTOR; MUSCARINIC ACETYLCHOLINE-RECEPTOR; SMOOTH-MUSCLE CELLS; B2 KININ RECEPTORS; BETA(2)-ADRENERGIC RECEPTOR; PLASMA-MEMBRANE; MEDIATED INTERNALIZATION; DISTINCT MECHANISMS; CAVEOLAE MEMBRANE; DOWN-REGULATION;
Keywords:
caveolae; clathrin-coated pit; G-protein-coupled receptor; internalization; peptide;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
44
Recensione:
Indirizzi per estratti:
Indirizzo: Leeb-Lundberg, LMF Univ Texas, Hlth Sci Ctr, Dept Biochem, 7703 Floyd CurlDr, San Antonio, TX 78229 USA Univ Texas 7703 Floyd Curl Dr San Antonio TXUSA 78229
Citazione:
M.E. Lamb et al., "Agonist-promoted trafficking of human bradykinin receptors: arrestin- and dynamin-independent sequestration of the B-2 receptor and bradykinin in HEK293 cells", BIOCHEM J, 355, 2001, pp. 741-750

Abstract

In this study, we analysed the agonist-promoted trafficking of human B-2 (B2R) and B-1 (B1R) bradykinin (BK) receptors. using wild-type End green fluorescent protein (GFP)-tagged receptors in HEK293 cells. B2R was sequestered to a major extent upon exposure to BK, as determined by the loss of cell-surface B2R using radioligand binding and by imaging of B2R-GFP using laser-scanning confocal fluorescence microscopy. Concurrent BK sequestration wasrevealed by the appearance of acid-resistant specific BK receptor binding. The same techniques showed that B1R was sequestered to a considerably lesser extent upon binding of des-Arg(10)-kallidin. B2R sequestration was rapid(half-life similar to5 min) and reached a steady-state level that was significantly lower than that of BK sequestration. B2R sequestration was minimally inhibited by K44A dynamin (22.4 +/- 3.7%,), and was insensitive to arrestin-(319-418), which are dominant-negative mutants of dynamin I and beta -arrestin respectively. Furthermore, the B2R-mediated sequestration of BK was completely insensitive to both mutants, as was the association of BK witha caveolae-enriched fraction of the cells. On the other hand, agonist-promoted sequestration of the beta (2)-adrenergic receptor was dramatically inhibited by K44A dynamin (81.2 +/- 16.3%) and by arrestin-(319-418) (36.9 +/-4.4%). Our results show that B2R is sequestered to a significantly greaterextent than is B2R upon agonist treatment in HEK293 cells. Furthermore, B2R appears to be recycled in the process of sequestering BK, and this process occurs in a dynamin- and beta -arrestin-independent manner and, at least in part,. involves caveolae.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 21/09/20 alle ore 11:57:44