Glucagon receptor proteins, peptides, and antibodies
First Claim
1. An isolated and purified polypeptide comprising the amino acid sequence of a glucagon receptor having seven transmembrane domains, an N-terminal extracellular domain, a C-terminal intracellular domain, three extracellular loop domains, and three intracellular loop domains,wherein the receptor is capable of binding glucagon with a KD no greater than 100 nM and is capable of transducing signal in a suitable host cell in response to such binding,and wherein the receptor is encoded by DNA which hybridizes under conditions of high stringency to a probe having the nucleotide sequence of the complement of nucleotides 145-1599 of SEQ ID NO:
- 14 or nucleotides 53-1483 of SEQ ID NO;
24.
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Abstract
The present invention provides isolated DNA molecules comprising a DNA segment encoding a glucagon receptor. Also provided are DNA constructs comprising a first DNA segment encoding a glucagon receptor operably linked to additional DNA segments required for the expression of the first DNA segment, as well as host cells containing such DNA constructs. The present invention also provides a method for detecting the presence of glucagon antagonists, comprising the steps of (a) exposing a compound in the presence of a glucagon against to a recombinant glucagon receptor coupled to a response pathway under conditions and for time sufficient to allow binding of the compound to the receptor and an associated response through the pathway, and (b) detecting a reduction in the stimulation of the response pathway resulting from the binding of the compound to the glucagon receptor, relative to the stimulation of the response pathway by the glucagon agonist alone and therefrom determining the presence of a glucagon antagonist.
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Citations
18 Claims
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1. An isolated and purified polypeptide comprising the amino acid sequence of a glucagon receptor having seven transmembrane domains, an N-terminal extracellular domain, a C-terminal intracellular domain, three extracellular loop domains, and three intracellular loop domains,
wherein the receptor is capable of binding glucagon with a KD no greater than 100 nM and is capable of transducing signal in a suitable host cell in response to such binding, and wherein the receptor is encoded by DNA which hybridizes under conditions of high stringency to a probe having the nucleotide sequence of the complement of nucleotides 145-1599 of SEQ ID NO: - 14 or nucleotides 53-1483 of SEQ ID NO;
24. - View Dependent Claims (2, 5, 6, 7, 8, 12, 13, 14, 15, 17)
- 14 or nucleotides 53-1483 of SEQ ID NO;
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3. An isolated and purified polypeptide comprising the amino acid sequence of a glucagon receptor native to a mammal, the receptor having seven transmembrane domains, an N-terminal extracellular domain, a C-terminal intracellular domain, three extracellular loop domains, and three intracellular loop domains,
wherein the receptor is encoded by DNA which hybridizes under conditions of high stringency to a probe having the nucleotide sequence of the complement of nucleotides 145-1599 of SEQ ID NO: - 14 or nucleotides 53-1483 of SEQ ID NO;
24. - View Dependent Claims (4)
- 14 or nucleotides 53-1483 of SEQ ID NO;
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9. A peptide comprising the amino acid sequence of a fragment of a glucagon receptor polypeptide having the sequence shown as residues 28-485 of SEQ ID NO:
- 15, residues 1-485 of SEQ ID NO;
15, or residues 1-477 of SEQ ID NO;
25, the fragment comprising at least(a) the complete sequence of one of the extracellular or intracellular domains of the receptor, or (b) ten contiguous amino acids from the sequence of the receptor. - View Dependent Claims (10, 11, 16, 18)
- 15, residues 1-485 of SEQ ID NO;
Specification