[Objective]The experiment was to study the interaction between chicken interferonγ(chIFN-γ)and chicken complement receptor 2(chCR2)proteins,and lay a foundation for the research of new functions of chIFN-γ and chCR2.[Method]chIFN-γ related expression plasmid was constructed by homologous recombination method.The interaction between chIFN-γ and chCR2 proteins was identified by co-immunoprecipitation,laser confocal and surface plasmon resonance techniques,and the interaction mode of chIFN-γ and chCR2 proteins was simulated by molecular docking.[Result]The pCMV-Myc-chIFN-γ expression plasmid was constructed successfully.The plasmid of pCMV-Myc-chIFN-γ and pCMV-HA-chCR2-ΔTM were co-transfected into HEK-293FT cells and the results showed that chIFN-γ could interact with chCR2.The plasmid of pCMV-Myc-chIFN-γ and pCMV-HA-chCR2-ΔTM were co-transfected into DF-1 cells and the results showed that chIFN-γ and chCR2 proteins could co-locate in DF-1 cells.The results of surface plasmon resonance test showed that the equilibrium dissociation constant(KD)between chIFN-γ and chCR2 proteins was 0.362 μmol/L,and the affinity between the two proteins was high.The results of molecular docking experiments showed that the interaction between chIFN-γ and chCR2 proteins was 1 salt bridge between Asp23-His150 and 3 sets of hydrogen bonds between Asn37-Ser143,Glu57-Gly179 and Ile93-Tyr184.[Conclusion]The results of this study proved that chIFN-γ and chCR2 proteins could interact,and the interaction mode between them was 1 salt bridge and 3 sets of hydrogen bonds.
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