Cloned glucagon-like peptide-2 receptors
View Patent ↗The invention relates to nucleotides and amino acid sequences encoding glucagon-like peptide 2 receptors, recombinant host cells transformed with such nucleotides, and methods of using the same in drug screening and related applications.
1. An isolated polynucleotide which encodes a GLP-2 receptor selected from:
(a) a human GLP-2 receptor comprising the amino acid sequence of amino acids 67-553 of SEQ ID NO: 12; and
(b) a GLP-2 receptor which is at least 95% identical to amino acids 26-553 of SEQ ID NO: 12 and which exhibits the functional characteristic of selectively binding GLP-2.
2. An isolated polynucleotide which encodes a GLP-2 receptor according to claim 1 wherein said GLP-2 receptor is a human GLP-2 receptor comprising the amino acid sequence of amino acids 67-553 of SEQ ID NO: 12.
3. An isolated polynucleotide according to claim 1 , comprising nucleotides 320-1780 of SEQ ID NO: 11.
4. An isolated polynucleotide according to claim 1 , wherein said polynucleotide encodes a variant of said human GLP-2 receptor, said variant comprising a substitution of Arg85.
5. An isolated polynucleotide according to claim 4 , which encodes a Glu85 variant.
6. An isolated polynucleotide according to claim 1 , wherein said polynucleotide encodes for the human GLP-2 receptor of amino acids 26-553 of SEQ ID NO: 12.
7. An isolated polynucleotide according to claim 1 , wherein said polynucleotide encodes for the human GLP-2 receptor and said polynucleotide encodes for the amino acid sequence which is at least 95% identical to amino acids 26-553 of SEQ ID NO: 12.
8. The polynucleotide of claim 1 , further comprising a label.
9. A recombinant polynucleotide comprising a GLP-2 receptor-encoding polynucleotide as defined in claim 1 , and expression controlling elements linked operably therewith to drive expression thereof.
10. A cell that has been genetically engineered by the incorporation expressibly therein of a polynucleotide according to claim 1 .
11. The cell according to claim 9 , which is a mammalian cell.
12. An isolated polynucleotide according to claim 1 , wherein said nucleotide sequence has at least 95% sequence identity to nucleotides 320-1780 of SEQ ID NO: 11, wherein said polynucleotide encodes a mammalian GLP-2 receptor that exhibits the functional characteristics of selectively binding GLP-2.
13. A method for identifying GLP-2 receptor ligands comprising:
(a) incubating a candidate ligand with a cell as defined in claim 11 or with a membrane preparation obtained therefrom, and then
(b) determining whether binding between the GLP-2 receptor and the candidate ligand has occurred.
14. A method for identifying GLP-2 receptor ligands, comprising the steps of:
(a) identifying a cell expressing a functional GLP-2 receptor comprising the amino acid sequence of amino acids 67-553 of SEQ ID NO: 12,
(b) incubating a candidate ligand with the cell that expresses a functional GLP-2 receptor, or with a membrane preparation derived from said cell; and
(c) determining whether binding between the GLP-2 receptor and the ligand has occurred.
15. A method according to claim 14 , wherein the candidate ligand is incubated with a cell that produces a functional GLP-2 receptor, and the determination of whether binding has occurred between the GLP-2 receptor and the candidate ligand is achieved by measuring change in the intracellular cAMP level, wherein an increase in the cAMP level indicating that the candidate ligand is a GLP-2 agonist.