A soluble binding assay for measuring 3H-FK506 binding to the hsp56 immunophilin.
J Immunoassay
; 15(4): 339-55, 1994 Nov.
Article
en En
| MEDLINE
| ID: mdl-7530727
Heat shock protein 56 (hsp56) was previously identified as an immunophilin based on its ability to specifically bind to FK506-Affi-Gel 10. In this report, we have quantitated human Jurkat T cell hsp56 binding to 3H-FK506, as well as to the immunosuppressant rapamycin. Binding was measured utilizing immunoadsorbed hsp56, and, in addition, we demonstrate that 3H-FK506 binds to hsp56 in solution. Hsp56 bound to an antibody-Sepharose column binds 3H-FK506 with an affinity of 19.4 +/- 4.6 nM, as compared to 23.2 +/- 6.8 nM for soluble hsp56. In competition experiments, the apparent affinity constant for rapamycin was 11.6 +/- 2.8 nM, using immobilized hsp56, and 17.3 +/- 7.7 nM, using the soluble hsp56 preparation. These results demonstrate that hsp56 binds FK506 and rapamycin with similar affinities, and suggest that hsp56 may play a role in mediating the cellular function of both of these drugs.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Polienos
/
Proteínas Portadoras
/
Cromatografía de Afinidad
/
Tacrolimus
/
Proteínas de Unión al ADN
/
Proteínas de Choque Térmico
Límite:
Humans
Idioma:
En
Revista:
J Immunoassay
Año:
1994
Tipo del documento:
Article
Pais de publicación:
Estados Unidos