Histamine receptor
View Patent ↗The present invention provides an isolated mammalian histamine receptor, isolated or recombinant nucleic acids and recombinant vectors encoding the same, host cells comprising the nucleic acids and vectors, and methods of making the receptor using the host cells. This invention further provides antibodies and antigen binding fragments thereof which specifically bind to the receptor and are useful for treating medical conditions caused or mediated by histamine. Also provided are screening methods for identifying specific agonists and antagonists of the mammalian histamine receptor.
1. A method for identifying an agonist or antagonist of a mammalian histamine receptor, comprising:
(a) contacting a polypeptide comprising the amino acid sequence defined by SEQ ID NO: 2 or a histamine binding fragment thereof in the presence of a known amount of labeled histamine or a surrogate thereof with a sample to be tested for the presence of a histamine agonist or antagonist;
(b) measuring the amount of labeled histamine or surrogate thereof specifically bound to the polypeptide;
(c) contacting cells expressing a polypeptide comprising the amino acid sequence defined by SEQ ID NO: 2 or a histamine binding fragment thereof with said sample; and
(d) measuring at least one cellular function modulated by the binding of a ligand to a polypeptide comprising the amino acid sequence defined by SEQ ID NO: 2;
whereby a histamine agonist in the sample is identified by measuring:
(i) substantially reduced binding of the labeled histamine or surrogate thereof to the polypeptide compared to what would be measured in the absence of said sample in step (b); and
(ii) substantially increased level of said cellular function compared to what would be measured in the absence of said sample in step (d);
and whereby a histamine antagonist in the sample is identified by measuring:
(i) substantially reduced binding of the labeled histamine or surrogate thereof to the polypeptide compared to what would be measured in the absence of said sample in step (b); and
(ii) substantially similar or reduced level of said cellular function compared to what would be measured in the absence of said sample in step (d).
2. The method of claim 1 , wherein said at least one cellular function is selected from the group consisting of:
(a) inhibition of forskolin-stimulated intracellular cAMP production;
(b) Ca 2+ mobilization;
(c) inositol phosphate production; and
(d) intracellular release of calcium.