Chimeric protein for the screening of agonists and antagonists of cell signalling pathways that are dependent on g-protein-coupled receptors

摘要

Chimeric protein (I) derived from a high-threshold calcium channel is new. It consists of at least one beta-subunit (Ia), or its fragment that includes the interaction domain BID, fused at either end with the I-II loop (Ib) of the alpha1 subunit, or fragment of it containing the interaction domain AID. Independent claims are also included for the following: (1) variant chimeric protein (II) derived from (I) and having at least one amino acid (aa) mutation in either (Ia) and (Ib); (2) peptide (III) that is a fragment of at least 7 aa from (I) or (II), situated at the beta/alpha junction; (3) antibodies (Ab) directed against (I)-(II); (4) nucleic acid (IV) that encodes (I)-(III), also their complements, sense or antisense; (5) probes and primers that contain a sequence of 10-30 nt corresponding to the beta/alpha1 junction of (I); (6) recombinant vector that contains (IV); (7) cells modified by the vector of (6), by (IV) or by (I) or (II); (8) transgenic non-human mammal in which at least some cells are transfected by (IV); (9) method for studying signaling and regulatory processes in cells that are dependent on G protein-coupled receptors; (10) method of screening for (ant)agonists of such signaling and regulatory processes; (11) method of screening for antagonists of the interaction between the alpha1 and beta subunits of high-threshold calcium channels; and (12) kits for performing methods (9)-(11).