| 基于表面等离子体共振(SPR)技术实时分析RGD基序与整联蛋白的相互作用 |
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| 引用本文： | 李鹏飞,龚真莉,杜晓华,蒋韬.基于表面等离子体共振(SPR)技术实时分析RGD基序与整联蛋白的相互作用[J].畜牧兽医学报,2021,52(1):256-261. |
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| 作者姓名： | 李鹏飞 龚真莉 杜晓华 蒋韬 |
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| 作者单位： | 1. 甘肃农业大学 动物医学院, 兰州 730070;2. 中国农业科学院 兰州兽医研究所 家畜疫病病原生物学国家重点实验室 国家口蹄疫参考实验室, 兰州 730046 |
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| 基金项目： | 国家自然科学基金(31772717)。 |
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| 摘 要： | 旨在研究口蹄疫病毒（foot-and-mouth disease virus，FMDV）结构蛋白VP1上的RGD（Arg-Gly-Asp）基序与宿主细胞表面受体整联蛋白的结合特异性，作者应用基于表面等离子共振技术（SPR）的Biacore 3000系统实时研究RGD基序分别与猪源整联蛋白αvβ6胞外区结构域、αv亚基胞外区结构域和β6亚基胞外区结构域的亲和力。首先通过结合试验筛选与RGD基序有结合的整联蛋白结构域，再对有结合的整联蛋白与RGD基序开展动力学分析。结果显示，合成的RGD基序与猪源整联蛋白αvβ6胞外区结构域有结合，结合动力学常数Ka、Kd、KD分别为42.3 M-1s-1、3.1×10-4s-1和7.33×10-6M；与整联蛋白αv亚基胞外区结构域之间亦有结合，结合动力学常数Ka、Kd、KD分别为21.8 M-1s-1、2.13×10-4s-1和9.79×10-5M；与β6亚基胞外区结构域几乎没有结合。综上表明，RGD与整联蛋白αvβ6胞外区结构域的结合比与整联蛋白αv亚基胞外区结构域之间的结合快且亲和力强。本研究将为进一步探讨FMDV与宿主细胞表面受体的相互作用提供参考。
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| 关 键 词： | Biacore 3000 FMDV RGD基序 表面等离子体共振 整联蛋白 动力学 |
| 收稿时间： | 2020-06-12 |
Real-time Analysis of the Interaction between RGD Motif and Integrin Based on Surface Plasmon Resonance (SPR) |
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| LI Pengfei,GONG Zhenli,DU Xiaohua,JIANG Tao.Real-time Analysis of the Interaction between RGD Motif and Integrin Based on Surface Plasmon Resonance (SPR)[J].Acta Veterinaria et Zootechnica Sinica,2021,52(1):256-261. |
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| Authors: | LI Pengfei GONG Zhenli DU Xiaohua JIANG Tao |
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| Institution: | 1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;2. Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China |
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| Abstract: | This experiment aims to study the binding specificity of RGD (Arg-Gly-Asp) motif on the structural protein VP1 of foot-and-mouth disease virus (FMDV) to host cell surface receptor integrin. In this paper, the Biacore 3000 system based on surface plasmon resonance (SPR) was used to study the binding specificity of RGD motif with the αvβ6 extracellular domain, αv extracellular domain and β6 extracellular domain, respectively. Firstly, the domains of integrin bound to RGD Motif were screened by binding experiments, and then the kinetic analysis of integrin bound to RGD Motif was carried out. The results showed that the synthesized RGD motif was bound to the αvβ6 extracellular domain, and the kinetic constants Ka, Kd, and KD were 42.3 M-1s-1, 3.1×10-4s-1, and 7.33×10-6M respectively. It also binds to the αv extracellular domain, the kinetic constants were 21.8 M-1s-1, 2.13×10-4s-1,and 9.79×10-5M respectively. There was almost no binding to the extracellular domain of the β6 subunit. The results showed that the binding of RGD to the extracellular domain of αvβ6 was faster and stronger than that of the extracellular domain of αv subunit. This study will provide a reference for further study of the interaction between FMDV and host cell surface receptors. |
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| Keywords: | Biacore 3000 FMDV RGD motif surface plasmon resonance integrin kinetics |
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