Researching Homologous Dimerization of G Protein Coupling Receptor PAC1 through BRET and BiFC

Abstract: G-protein couple receptors (GPCRs) are the biggest super family membrane receptors. PAC1 belongs to the B family of GPCRs and is pituitary adenosine acid cyclization enzyme excited peptide (PACAP) specific receptors， mediating PACAP neural protection function， which is one of the important targets for drug development to diseases of the nervous system. Dimerization or oligomerization is a common phenomenon to GPCRs. But there is no report homologus dimerization or oligomerization for PAC1 at present. In order to verify PAC1 dimerization， we use BRET to test CHO cells which are co-transfected PAC1-Rluc and PAC1-EYFP with different density gradient. The result presents obviously BRET signal by adding coelenterazine h. While BiFC test shows that CHO cells which are co-transfecte PAC1-EYFP/N and PAC1-EYFP/C appear complete EYFP fluorescent signal. Western blot test also shows that cells which high expressing PAC1 contain macromolecules of PAC1 dimer. So PAC1 can normally form homologous dimerization. This discovery will lay the novel theoretical foundation for the subsequent drug development， and offer illumination and reference for researching other GPCRs.

CSTR: 32200.14.cjcb.2012.10.0011