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Titolo:
Reconstitution of human 5-hydroxytryptamine(5A) receptor-G protein coupling in E-coli and Sf9 cell membranes with membranes from Sf9 cells expressingmammalian G proteins
Autore:
Francken, BJB; Vanhauwe, JFM; Josson, K; Jurzak, M; Luyten, WHML; Leysen, JE;
Indirizzi:
Janssen Res Fdn, Dept Biochem Pharmacol, B-2340 Beerse, Belgium Janssen Res Fdn Beerse Belgium B-2340 Pharmacol, B-2340 Beerse, Belgium Janssen Res Fdn, Dept Funct Genom, B-2340 Beerse, Belgium Janssen Res FdnBeerse Belgium B-2340 unct Genom, B-2340 Beerse, Belgium
Titolo Testata:
RECEPTORS & CHANNELS
fascicolo: 4, volume: 7, anno: 2001,
pagine: 303 - 318
SICI:
1060-6823(2001)7:4<303:ROH5RP>2.0.ZU;2-7
Fonte:
ISI
Lingua:
ENG
Soggetto:
5-HT5A SEROTONIN RECEPTOR; ESCHERICHIA-COLI; ADENYLATE-CYCLASE; ALPHA-SUBUNITS; FUNCTIONAL EXPRESSION; REGULATORY COMPONENT; BINDING-PROPERTIES; LIGAND-BINDING; HEK-293 CELLS; HIGH-AFFINITY;
Keywords:
Escherichia coli; expression; G protein; human 5-ht(5A) receptor; reconstitution; Sf9 insect cell;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
37
Recensione:
Indirizzi per estratti:
Indirizzo: Leysen, JE Janssen Pharmaceut, Turnhoutseweg 30, B-2340 Beerse, Belgium Janssen Pharmaceut Turnhoutseweg 30 Beerse Belgium B-2340 gium
Citazione:
B.J.B. Francken et al., "Reconstitution of human 5-hydroxytryptamine(5A) receptor-G protein coupling in E-coli and Sf9 cell membranes with membranes from Sf9 cells expressingmammalian G proteins", RECEPT CHAN, 7(4), 2001, pp. 303-318

Abstract

The human 5-hydroxytryptamine(5A) (h5-ht(5A)) receptor was expressed in Escherichia coli (hS-ht(5A)-E. coli) to verify its pharmacological profile inthe absence of G proteins. In addition, the ability of the h5-ht(5A) receptor to interact with mammalian G(i/o), and G(s) proteins was investigated by a new reconstitution approach. Agonists displayed lower affinities for h5-ht(5A)-E. coli than for stably transfected h5-h(5A),-HEK 293 cells, due tothe absence of G protein coupling in E. coli, Lysergic acid diethylamide behaved as a neutral antagonist, showing equal affinities for the G protein-coupled and the uncoupled receptor. To analyze the G protein coupling behavior of the h5-ht(5A) receptor, h5-ht(5A)-E. coli membranes or h5-ht(5A)-Sf9insect cell membranes were fused by vortexing to membranes from baculovirus-infected Sf9 cells expressing mammalian G proteins. The ability of the h5-ht(5A), receptor to differentiate between G(i)/G(o)/G(z), and G(s) proteins was explored by investigation of agonist binding affinities and agonist-induced stimulation of [S-35]GTP gammaS binding. The h5ht(5A), receptor failed to interact with G(z) and G(s) proteins and coupled equally well to G(i)and G(o) proteins to form a complex with high affinity for agonists. Underthe applied conditions, however, G(i) proteins were found to be better activated than G(o) proteins in the [35S]GTP gammaS binding assay.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 20/01/20 alle ore 10:46:28