Study on the binding ability of gD protein mutation of PRV-2022 strain to human Nectin-1
10.3760/cma.j.cn112866-20240412-00060
- VernacularTitle:伪狂犬病毒PRV-2022毒株gD蛋白突变与人Nectin-1结合力
- Author:
Xinyue WANG
1
;
Weiyu WANG
;
Guoyong MEI
;
Jun HAN
;
Cao CHEN
Author Information
1. 中国疾病预防控制中心病毒病预防控制所 传染病溯源预警与智能决策全国重点实验室 国家卫生健康委医学病毒和病毒病重点实验室，北京 102206
- Keywords:
Pseudorabies;
Pseudorabies virus;
Glycoprotein D;
Nectin-1 receptor;
Molecular interaction
- From:
Chinese Journal of Experimental and Clinical Virology
2024;38(4):395-401
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the impact of various mutations in the gD protein of PRV-2022 strain on its binding to the Nectin-1 receptor.Methods:We employed PCR, RT-qPCR and gene sequencing techniques for identification of the PRV-2022 strain. Furthermore, bioinformatics method were utilized to analyze the genetic evolution of the gD gene in PRV-2022 strain. Recombinant expression plasmid containing mutations at amino acids positions 69 and 82 within the extracellular domain of gD protein from PRV-2022 strain was constructed and expressed in vitro. The binding ability between different mutant forms of recombinant gD protein and Nectin-1 receptor was compared using His-pull down and biolayer interference techniques. Results:The gD gene of the PRV-2022 strain was obtained, and genetic evolution analysis revealed that the PRV-2022 strain belonged to the same branch as strains isolated prior to 2011, with a close genetic distance. The expression plasmids for gD extracellular domain containing A69V and S82N amino acid mutations were successfully constructed, enabling the expression and purification of recombinant PRV gD extracellular domain protein. Interaction studies demonstrated that gD-69, gD-82, gD-2022, and gD-Bartha proteins interacted with human Nectin-1. Notably, compared to the classical PRV vaccine strain Bartha, double mutation of amino acids 69 and 82 in the gD protein exhibited the highest affinity to human Nectin-1 receptor, whereas individual mutations at either site decreased this affinity.Conclusions:Introduction of A69V and S82N mutations in the gD protein significantly affected its binding ability to human Nectin-1 receptor. Simultaneous occurrence of A69V and S82N mutations resulted in the highest affinity towards human Nectin-1 receptor.
