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Titolo:
STRUCTURE-FUNCTION RELATIONSHIP OF DIFFERENT DOMAINS OF THE RAT CORTICOTROPIN-RELEASING FACTOR-RECEPTOR
Autore:
SYDOW S; RADULOVIC J; DAUTZENBERG FM; SPIESS J;
Indirizzi:
MAX PLANCK INST EXPT MED,DEPT MOL NEUROENDOCRINOL,HERMANN REIN STR 3 D-37075 GOTTINGEN GERMANY
Titolo Testata:
Molecular brain research
fascicolo: 2, volume: 52, anno: 1997,
pagine: 182 - 193
SICI:
0169-328X(1997)52:2<182:SRODDO>2.0.ZU;2-7
Fonte:
ISI
Lingua:
ENG
Soggetto:
Y-79 RETINOBLASTOMA CELLS; CHEMICAL CROSS-LINKING; CENTRAL-NERVOUS-SYSTEM; FACTOR-BINDING-PROTEIN; FACTOR CRF RECEPTORS; HORMONE RECEPTORS; PLASMA-MEMBRANE; MOUSE PITUITARY; BRAIN; IDENTIFICATION;
Keywords:
CORTICOTROPIN-RELEASING FACTOR RECEPTOR; CRFR1 ANTIBODIES; HEK 293 CELL; CHO-K1 CELL; HISTIDINE TAG; N-GLYCOSYLATION; CRF BINDING; CYCLIC AMP;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
46
Recensione:
Indirizzi per estratti:
Citazione:
S. Sydow et al., "STRUCTURE-FUNCTION RELATIONSHIP OF DIFFERENT DOMAINS OF THE RAT CORTICOTROPIN-RELEASING FACTOR-RECEPTOR", Molecular brain research, 52(2), 1997, pp. 182-193

Abstract

The significance of different domains of corticotropin-releasing factor receptor, type I, (CRFR1) for ligand binding and cAMP accumulation was investigated with C-terminally truncated forms of rat CRFR1 (rCRFR1) tagged by a sequence of six histidine residues (His-tag) to facilitate protein purification and identification. These different forms of the receptor were N-glycosylated and transported properly to the membranes of transfected mammalian cells as indicated by Western blot analysis and immunocytochemical staining with two polyclonal antibodies developed against the N- and C-terminus of rCRFR1. The N-terminal fragment, rCRFR1(23-121), expressed in Escherichia coli bound oCRF specifically, but with low affinity. Several mutants lacking transmembrane domain (TM) 7 and the C-terminus exhibited similarly low affinities to oCRFafter expression in transfected mammalian cells. None of these cells produced significant amounts of cAMP after exposure to oCRF. Only mutants containing the N-terminus. all loops and TMs bound oCRF and produced cAMP with high affinity (K-d = 62 nM) and efficacy (EC50 = 0.8 nM). The additional presence of the C-terminus provided similar characteristics (K-d = 5 nM, EC50 = 0.3 nM) as known for the native receptor. Itis suggested on the basis of these data that the last extracellular loop is involved in ligand binding. (C) 1997 Elsevier Science B.V.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 22:34:40