Relaxin3-gpcr135 complexes and their production and use
OwnerJingcai Chen, Chester Kuei +4inventors
InventorsJingcai Chen, Chester Kuei, Changlu Liu, Timothy Lovenberg, Rannar Sillard, Steven Sutton
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Abstract
A GPCR135/relaxin3 complex is described, which is useful as an assay reagent for screening for modulators of GPCR135 receptor activity.
Claims30
A receptor-ligand complex comprising a receptor component containing GPCR135 or an active fragment of GPCR135 bound to a ligand component containing relaxin3 or an active fragment of relaxin3, wherein at least one of the receptor and ligand components is in a substantially pure form.
An isolated polynucleotide having a nucleotide sequence encoding an amino acid sequence of SEQ ID NO:
- 15, or a complement thereof.
An isolated polypeptide having an amino acid sequence of SEQ ID NO:
- 15.
A vector consisting of a polynucleotide encoding a polypeptide having an amino acid sequence of SEQ ID NO:
- 15.
A recombinant host cell comprising a vector comprising a polynucleotide encoding a polypeptide having an amino acid sequence of SEQ ID NO:
- 15.
A process of producing a relaxin3 from a recombinant cell, comprising:
- (a) constructing a DNA molecule encoding a relaxin3 propeptide modified by insertion of a peptide linkage forming a protease cleavage site at at least one location selected from the peptide junction between chains A and C and the peptide junction between chains C and B of the relaxin3 propeptide;
- (b) constructing a first vector expressing the modified relaxin3 propeptide;
- (c) constructing a second vector expressing a protease for cleaving the modified relaxin3 propeptide at the inserted protease cleavage site;
- (d) introducing both the first vector and the second vector into a host cell;
- and
- (e) growing the host cell so that both the modified relaxin3 propeptide and the protease are expressed, whereby the protease efficiently cleaves the peptide linkage at said location on the modified relaxin3 propeptide.
A method of identifying a compound that binds to GPCR135 or an active fragment thereof, comprising the steps of:
- (a) contacting GPCR135 or an active fragment thereof with a test compound and with a labeled relaxin3 or an active fragment thereof;
- (b) determining the amount of the labeled relaxin3 or active fragment thereof that binds to the GPCR135 or active fragment thereof;
- and
- (c) comparing the amount determined in step (b) with a control measurement wherein the GPCR135 or active fragment thereof has been contacted with the labeled relaxin3 or active fragment thereof in the absence of test compound.
A method for identifying a compound that binds GPCR135 and mimics relaxin3, comprising:
- (a) contacting a test compound with an assay reagent comprising GPCR135 or an active fragment thereof;
- (b) determining a biological activity of the GPCR135 or active fragment thereof;
- and
- (c) comparing the result determined in step (b) with that of a control measurement wherein the GPCR135 or an active fragment thereof was contacted with relaxin3 or an active fragment thereof in the absence of the test compound.
Dependencies are inferred from the claim text.