Human dopamine receptor and its uses
AssigneeOregon Health Sciences Univ.
InventorsOlivier Civelli, Hubert Henri-Marie Van Tol
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Abstract
The present invention is directed toward isolation, characterization and pharmacological use of the human D4 dopamine receptor. The nucleotide sequence of the gene corresponding to this receptor and alleleic variant thereof are provided by the invention. The invention also includes recombinant eukaryotic expression constructs capable of expressing the human D4 dopamine receptor in cultures of transformed eukaryotic cells. The invention provides cultures of transformed eukaryotic cells which synthesize the human D4 dopamine receptor, and methods for characterizing novel psychotropic compounds using such cultures.
Claims12
A method of screening a compound for binding to a human D4 dopamine receptor, the method comprising the following steps:
- (a) transforming a host cell with a recombinant expression construct encoding a human D4 dopamine receptor having an amino acid sequence that is the amino acid sequence of the human D4 dopamine receptor allele D4.4 from amino acid 1 to amino acid 248 (SEQ ID No.:
- 20), covalently linked at the carboxyl terminus of amino acid 248 to the amino terminus of a repeated amino acid sequence, the carboxyl terminus of the repeated amino acid sequence being covalently linked to the amino terminus of the amino acid sequence of the human D4 dopamine receptor allele D4.4 from amino acid 312 to amino acid 419 (SEQ ID No.:
- 20), wherein the amino acid sequence of the repeated sequence comprises from 3 to 8 copies of the amino acid sequence:
- (Pro/Ala).Ala.Pro.(Arg/Gly).Leu.Pro.(Gln/Arg/Pro).(Asp/Gly).Pro.Cys.Gly.(Pro/Ser).(Asp/Asn).Cys.Ala.Pro (SEQ ID No.:24),
- wherein the cells of the transformed cell culture express the receptor;
- and
- (b) assaying the transformed cell with the compound to determine whether the compound binds to the human D4 dopamine receptor.
A method of screening a compound for competitive binding to a human D4 dopamine receptor, the method comprising the following steps:
- (a) transforming a host cell with a recombinant expression construct encoding a human D4 dopamine receptor having an amino acid sequence that is the amino acid sequence of the human D4 dopamine receptor allele D4.4 from amino acid 1 to amino acid 248 (SEQ ID No.:
- 20), covalently linked at the carboxyl terminus of amino acid 248 to the amino terminus of a repeated amino acid sequence, the carboxyl terminus of the repeated amino acid sequence being covalently linked to the amino terminus of the amino acid sequence of the human D4 dopamine receptor allele D4.4 from amino acid 312 to amino acid 419 (SEQ ID No.:
- 20), wherein the amino acid sequence of the repeated sequence comprises from 3 to 8 copies of the amino acid sequence:
- (Pro/Ala).Ala.Pro.(Arg/Gly).Leu.Pro.(Gln/Arg/Pro).(Asp/Gly).Pro.Cys.Gly.(Pro/Ser).(Asp/Asn).Cys.Ala.Pro (SEQ ID No.:24),
- wherein the cells of the transformed cell culture express the receptor;
- (b) assaying the transformed cell with the compound in the presence and in the absence of an agonist for the human D4 dopamine receptor;
- and
- (c) determining whether the compound competes with the agonist for binding to the human D4 dopamine receptor.
A method of screening a compound to determine if the compound is an agonist binding inhibitor of a human D4 dopamine receptor, the method comprising the following steps:
- (a) transforming a host cell with a recombinant expression construct encoding a human D4 dopamine receptor having an amino acid sequence that is the amino acid sequence of the human D4 dopamine receptor allele D4.4 from amino acid 1 to amino acid 248 (SEQ ID No.:
- 20), covalently linked at the carboxyl terminus of amino acid 248 to the amino terminus of a repeated amino acid sequence, the carboxyl terminus of the repeated amino acid sequence being covalently linked to the amino terminus of the amino acid sequence of the human D4 dopamine receptor allele D4.4 from amino acid 312 to amino acid 419 (SEQ ID No.:
- 20), wherein the amino acid sequence of the repeated sequence comprises from 3 to 8 copies of the amino acid sequence:
- (Pro/Ala).Ala.Pro.(Arg/Gly).Leu.Pro.(Gln/Arg/Pro).(Asp/Gly).Pro.Cys.Gly.(Pro/Ser).(Asp/Asn).Cys.Ala.Pro (SEQ ID No.:24),
- wherein the cells of the transformed cell culture express the receptor;
- and
- (b) assaying the transformed cell culture with the compound in the presence of a human D4 dopamine receptor agonist to determine whether the compound is capable of inhibiting agonist binding to the receptor.
Dependencies are inferred from the claim text.