Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
A yeast sensor of ligand binding
Autore:
Tucker, CL; Fields, S;
Indirizzi:
Univ Washington, Dept Genet, Seattle, WA 98195 USA Univ Washington Seattle WA USA 98195 n, Dept Genet, Seattle, WA 98195 USA Univ Washington, Dept Med, Seattle, WA 98195 USA Univ Washington Seattle WA USA 98195 ton, Dept Med, Seattle, WA 98195 USA Univ Washington, Howard Hughes Med Inst, Seattle, WA 98195 USA Univ Washington Seattle WA USA 98195 ghes Med Inst, Seattle, WA 98195 USA
Titolo Testata:
NATURE BIOTECHNOLOGY
fascicolo: 11, volume: 19, anno: 2001,
pagine: 1042 - 1046
SICI:
1087-0156(200111)19:11<1042:AYSOLB>2.0.ZU;2-#
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN FLUORESCENT PROTEINS; DIHYDROFOLATE-REDUCTASE; SACCHAROMYCES-CEREVISIAE; GENERIC BIOSENSORS; COMPLEXES; ESTROGEN; DOMAIN; COMPLEMENTATION; IDENTIFICATION; METHOTREXATE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
29
Recensione:
Indirizzi per estratti:
Indirizzo: Fields, S Univ Washington, Dept Genet, Box 357360, Seattle, WA 98195 USA Univ Washington Box 357360 Seattle WA USA 98195 le, WA 98195 USA
Citazione:
C.L. Tucker e S. Fields, "A yeast sensor of ligand binding", NAT BIOTECH, 19(11), 2001, pp. 1042-1046

Abstract

We describe a biosensor that reports the binding of small-molecule ligandsto proteins as changes in growth of temperature-sensitive yeast. The yeaststrains lack dihydrofolate reductase (DHFR) and are complemented by mouse DHFR containing a ligand-binding domain inserted in a flexible loop. Yeast strains expressing two ligand-binding domain fusions, FKBP12-DHFR and estrogen receptor-alpha (ER alpha)-DHFR, show increased growth in the presence of their corresponding ligands. We used this sensor to identify mutations inresidues of ER alpha important for ligand binding, as well as mutations generally affecting protein activity or expression. We also, tested the sensor against a chemical array to identify ligands that bind to FKBP12 or ER alpha. The ERa sensor was able to discriminate among estrogen analogs, showing different degrees of growth for the analogs that correlated with their relative binding affinities (RBAs). This growth assay provides a simple and inexpensive method to select novel ligands and ligand-binding domains.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 05/12/20 alle ore 02:01:14