Nuclear hormone receptor drug screens

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

Reexamination Certificate

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C434S007000, C434S007000

Reexamination Certificate

active

07101681

ABSTRACT:
Methods for identifying modulators of nuclear hormone receptor function comprise the steps of (a) forming a mixture comprising a nuclear hormone receptor, a peptide sensor and a candidate agent, but not a natural coactivator protein of the receptor, wherein the sensor provides direct, in vitro binding to the receptor under assay conditions; (b) measuring an agent-biased binding of the sensor to the receptor; and (c) comparing the agent-biased binding with a corresponding unbiased binding of the sensor to the receptor. In particular embodiments, the sensor comprises an amphipathic alpha helix nuclear hormone interacting domain comprising a recited nuclear hormone transcriptional coactivator motif sequence, the sensor is present at sub-micromolar concentration, the binding reaction occurs in solution, the sensor comprises a fluorescent label and the measuring step comprises detecting fluorescence polarization of the label. Reagents include labeled sensor peptides and reaction mixtures consisting essentially of nuclear hormone receptor, a peptide and a candidate agent.

REFERENCES:
patent: 6020141 (2000-02-01), Pantoliano et al.
patent: WO 9710337 (1997-03-01), None
Heery et al. (1997) Nature 387:733-736.
Torchia et al. (1997) Nature 387:677-684.
Montiminy (1997) Nature 387:654-655.
Mathis (1995) Clinical Chemistry 41:1391-1397, 1997.
Harlow et al. (1988) Antibodies, Cold Spring Harbor Laboratory New York, pp. 553-616, 1988.

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