Receptors: their identification, characterization, preparation a

Details Receptors: their identification, characterization, preparation a Receptors: their identification, characterization, preparation a

1991-11-25

1993-11-16

Hill, Jr., Robert J.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 694, 536 234, 536 235, 530399, C12Q 168, C12P 2106, C07K 1300, A61K 3724

Patent

active

052623009

ABSTRACT:
The present invention relates to bioassays for evaluating compounds as potential antagonists for members of the steroid hormone superfamily of receptor protein(s). The invention bioassay comprises culturing test cells expressing steroid receptor protein in the presence of increasing concentrations of at least one compound whose ability to inhibit the action of agonists on members of the steroid hormone superfamily of receptors is sought to be determined, and a fixed concentration of at least one agonist for said steroid hormone receptor protein(s). Test cells employed in the practice of the present invention contain DNA which expresses a member of the steroid hormone superfamily of receptor protein(s) in a form having improved trans-activation transcription activity, relative to wild-type receptor, and a DNA sequence comprising a response element for said hormone receptor protein operatively linked to a reporter gene. Improved trans-activation transcription activity is provided by receptor having a plurality of at least one parental receptor trans-activation domain selected from .tau..sub.1 or .tau..sub.2, located outside the DNA-binding and ligand-binding domains of the receptor protein. The invention assay is completed by monitoring the level of expression of reporter gene product as a function of the concentration of potential antagonist compound in the culture medium, thereby indicating the ability of potential antagonist compound(s) to inhibit activation of members of the steroid hormone superfamily of receptors.

REFERENCES:
patent: 5071773 (1991-12-01), Evans et al.
Giguere et al. 1986 , Cell 46:645-652.
Godowski et al. 1988 Science 241:812.

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