Identification of key binding sites of Rift Valley fever virus envelope protein Gn to receptor
10.13200/j.cnki.cjb.004614
- VernacularTitle:裂谷热病毒囊膜蛋白Gn与受体结合关键位点的鉴定
- Author:
Tianyi MA
1
Author Information
1. *Jianghan University School of Medicine, Wuhan Institute of Biomedical Sciences, Hubei Province Key Laboratory of Cognitive and Emotional Disorders, Wuhan 430056, Hubei Province, China
- Publication Type:Journal Article
- Keywords:
Rift Valley fever virus(RVFV);
Envelope protein Gn;
Receptor;
Antibody;
Competitive inhibition
- From:
Chinese Journal of Biologicals
2025;38(12):1422-1431
- CountryChina
- Language:Chinese
-
Abstract:
Objective To identify the key sites of Rift Valley fever virus(RVFV) envelope protein Gn binding to the receptor,in order to provide a basis for the development of RVFV infection inhibitors.Methods The eukaryotic cell expression system was used to express RVFV envelope protein Gn, RVFV positive antibodies(antibodies 140, 144, 268, R12 and R17),low density lipoprotein receptor-related protein 1(LRP1) receptor domains(ClusterⅡ-Fc and ClusterⅣ-Fc) and six Gn mutants(T173A, Q174A, D176A, K180A, S291A and K294A). ELISA method or competitive ELISA were used to detect the binding activity of LRP1 receptor domain and RVFV Gn, the effect of RVFV positive antibody on the binding ability of LRP1 receptor domain and RVFV Gn, and the effect of Gn point mutation on its binding ability to RVFV positive antibody and LRP1 receptor domain. The inhibitory effect of LRP1 domain on RVFV infection was detected by pseudovirus neutralization assay.Results ClusterⅡ-Fc and ClusteⅣ-Fc domains of LRP1 exhibited specific binding to RVFV Gn protein. The binding ability of antibodies 268, R12 and R17 to Gn protein was dose-dependent, and no significant binding activity was detected for antibodies 140 and 144. Antibodies 268, R12 and R17 showed a certain competitive inhibitory effect on the binding ability of ClusterⅡ-Fc to Gn protein, and the competitive effect of antibody 268 was the most significant. Only antibody 268 showed competitive effect on ClusterⅣ-Fc. The binding activity of Gn mutants to RVFV positive antibodies was lower than that of Gn. Except for Q174A, the binding ability of other Gn mutants to ClusterⅡ-Fc and ClusterⅣ-Fc was all lower than that of Gn. Among them, the binding ability of T173A, K180A, S291A and K294A to ClusterⅡ-Fc was significantly lower than that of Gn(F = 15. 83, P < 0. 05), and the binding ability of K180A, S291A and K294A to ClusterⅣ-Fc was significantly lower than that of Gn(F = 8. 75, P < 0. 05).Conclusion K180, S291 and K294 are the key amino acid sites for RVFV Gn protein to mediate the interaction with the host receptor LRP1, revealing part of the molecular basis of the binding of Gn to LRP1 and laying the foundation for the development of inhibitors targeting virus adsorption.
