Binding assay employing IGS4, a human G-protein coupled neuromedin receptor
The present invention relates to novel identified polynucleotides, polypeptides encoded by them and to the use of such- polynucleotides and polypeptides, and to their production. More particularly, the polynucleotides and polypeptides of the present invention relate to the G-protein coupled receptor family, referred to as G-family. The invention also relates to inhibiting or activating the action of such polynucleotides and polypeptides, to a vector containing said polynucleotides, a host cell containing such vector and transgenic animals where the IGS 4 -gene is either overexpressed, misexpressed, underexpressed or suppressed (knock-out animals). The invention further relates to a method for screening compounds capable to act as an agonist or an antagonist of said G-protein coupled receptor family IGS 4 . The invention also relates to the identification of the cognate ligand of the IGS 4 polypeptides of the invention. High affinity binding to said IGS 4 polypeptides is found for the neuropeptides known as neuromedin U.
1. A method of determining whether a substance is a potential ligand of a neuromedin receptor comprising:
(a) contacting cells expressing the receptor chosen from SEQ ID NO:2, SEQ ID NO:4 SEQ ID NO:6 or SEQ ID NO:8, or the polypeptide encoded by a DNA insert contained in the deposit no. CBS 102221 or the deposit no. CBS 102222 at the Centraalbureau voor Schimmelcultures at Baarn the Netherlands with labeled neuromedin U in the presence and in the absence of the substance;
or contacting a receptor membrane preparation comprising one of said receptors chosen from SEQ ID NO:2, SEQ ID NO:4 SEQ ID NO:6 or SEQ ID NO:8 or a polypeptide encoded by the DNA insert contained in the deposit no. CBS 102221 or the deposit no. CBS 102222 at the Centraalbureau voor Schimmelcultures at Baarn the Netherlands with labeled neuromedin U in the presence and in the absence of the substance; and
(b) measuring the binding of neuromedin U to the receptor.
2. A method of determining whether a substance is a potential ligand of a neuromedin receptor comprising:
(a) contacting a cell expressing the receptor chosen from SEQ ID NO:2, SEQ ID NO:4 SEQ ID NO:6 or SEQ ID NO:8, or the polypeptide encoded by a DNA insert contained in the deposit no. CBS 102221 or the deposit no. CBS 102222 at the Centraalbureau voor Schimmelcultures at Baarn the Netherlands with neuromedin U in the presence and in the absence of the substance; and
(b) determining whether intracellular calcium levels are modulated in the cell, wherein modulation of intracellular calcium levels indicates that the substance is a ligand of the receptor.