鸡γ-干扰素与补体受体2蛋白相互作用的研究

doi:10.16431/j.cnki.1671-7236.2024.10.028

摘要/Abstract

摘要： 【目的】研究鸡γ-干扰素(chicken interferon γ,chIFN-γ)与鸡补体受体2(chicken complement receptor 2,chCR2)蛋白的相互作用，为研究chIFN-γ和chCR2新功能奠定基础。【方法】通过同源重组方法构建chIFN-γ相关表达质粒，利用免疫共沉淀、激光共聚焦和表面等离子共振技术鉴定chIFN-γ与chCR2蛋白的相互作用，并通过分子对接技术模拟chIFN-γ与chCR2蛋白的相互作用模式。【结果】成功构建了pCMV-Myc-chIFN-γ表达质粒。将pCMV-Myc-chIFN-γ和pCMV-HA-chCR2-ΔTM质粒共转染HEK-293FT细胞，结果表明chIFN-γ蛋白可以与chCR2蛋白进行相互作用；将pCMV-Myc-chIFN-γ和pCMV-HA-chCR2-ΔTM质粒共转染DF-1细胞，结果表明chIFN-γ蛋白与chCR2蛋白可以在DF-1细胞中进行共定位。表面等离子共振试验结果显示chIFN-γ与chCR2蛋白之间的平衡解离常数(KD)值为0.362 μmol/L，二者之间有较高的亲和力。分子对接试验结果显示，chIFN-γ与chCR2蛋白之间的相互作用模式为Asp23-His150间形成1组盐桥作用，Asn37-Ser143、Glu57-Gly179和Ile93-Tyr184间形成3组氢键作用。【结论】本研究结果证明了chIFN-γ与chCR2蛋白可以进行相互作用，且二者之间的作用模式为1组盐桥和3组氢键作用。

关键词: 鸡γ-干扰素(chIFN-γ); 鸡补体受体2(chCR2); 相互作用; 亲和力

Abstract: 【Objective】 The experiment was to study the interaction between chicken interferon γ (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.

Key words: chicken interferon γ (chIFN-γ); chicken complement receptor 2 (chCR2); interaction; affinity

中图分类号:

S831

靳换, 孟昭英, 涂敏, 沈佳, 史爱华, 赵蕾, 习硕, 张建伟, 章振华. 鸡γ-干扰素与补体受体2蛋白相互作用的研究[J]. 中国畜牧兽医, 2024, 51(10): 4476-4484.

IN Huan, MENG Zhaoying, TU Min, SHEN Jia, SHI Aihua, ZHAO Lei, XI Shuo, ZHANGJianwei, ZHANG Zhenhua. Study on the Interaction Between Chicken Interferon γ and Complement Receptor 2 Protein[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(10): 4476-4484.

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