Claims
- 1. A method of screening a compound as an inhibitor of agonist binding to a mammalian adrenocorticotropic hormone receptor, the method comprising the following steps:
- (a) transforming a eukaryotic cell culture with an expression construct comprising a nucleotide sequence encoding a mammalian adrenocorticotropic hormone receptor having an amino acid sequence identified as SEQ ID NO:4 or SEQ ID NO:6, said construct being capable of expressing the adrenocorticotropic hormone receptor in a eukaryotic cell;
- (b) obtaining a membrane preparation from the transformed cells;
- (c) contacting the membrane preparations with the compound and a detectably labeled adrenocorticotropic hormone receptor agonist; and
- (d) assaying for ability of the compound to inhibit the binding of the detectably-labeled adrenocorticotropic hormone receptor agonist.
- 2. The method of claim 1 wherein the mammalian adrenocorticotropic hormone receptor is the bovine adrenocorticotropic hormone receptor.
- 3. The method of claim 1 wherein the mammalian adrenocorticotropic hormone receptor is the human adrenocorticotropic hormone receptor.
- 4. A method of assaying a compound to quantitatively characterize the compound as an inhibitor of agonist binding to a mammalian adrenocorticotropic hormone receptor, the method comprising the following steps:
- (a) transforming a eukaryotic cell culture with an expression construct comprising a nucleotide sequence encoding a mammalian adrenocorticotropic hormone receptor having an amino acid sequence identified as SEQ ID NO:4 or SEQ ID NO:6, said construct being capable of expressing the adrenocorticotropic hormone receptor in a eukaryotic cell;
- (b) obtaining a membrane preparation from the transformed cells;
- (c) contacting the membrane preparations with the compound and a detectably labeled adrenocorticotropic hormone receptor agonist; and
- (d) assaying for amount of a compound by measuring the extent of inhibition of binding of a detectably labeled adrenocorticotropic hormone receptor agonist.
- 5. The method of claim 4 wherein the mammalian adrenocorticotropic hormone receptor is the bovine adrenocorticotropic hormone receptor.
- 6. The method of claim 4 wherein the mammalian adrenocorticotropic hormone receptor is the human adrenocorticotropic hormone receptor.
- 7. The method of claim 4 wherein the compound to be tested is present in a human.
- 8. The method of claim 4 wherein the compound is present in human blood.
- 9. The method of claim 4 wherein the compound is present in human cerebrospinal fluid.
- 10. The method of claim 4 wherein the compound is unknown.
Parent Case Info
This is a divisional of application Ser. No. 08/077,673, filed on Jun. 15, 1993, now U.S. Pat. No. 5,554,729, issued Sep. 10, 1996, which is a divisional of Ser. No. 07/866,560, filed Apr. 10, 1992, now U.S. Pat. No. 5,280,112, issued Jan. 18, 1994.
Government Interests
This invention was made with government support under 1R01DK41921-03, 1R01DK43859-01, and 1P01DK44239-10A1 by the National Institutes of Health. The government has certain rights in the invention.
Non-Patent Literature Citations (6)
Entry |
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Divisions (2)
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Number |
Date |
Country |
Parent |
77673 |
Jun 1993 |
|
Parent |
866560 |
Apr 1992 |
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