Qian Lingling, Liu Yuwei, Wang Xiaochun, Yang Shixing, Ji Likai, Sun Xiaopeng, Wang Jianqiang, Shan Tongling, Zhang Wen, Shen Quan
Central Laboratory of Changshu Medicine Examination Institute, Changshu, Jiangsu, China.
Department of Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang, China.
Front Cell Infect Microbiol. 2025 May 13;15:1595736. doi: 10.3389/fcimb.2025.1595736. eCollection 2025.
Goose Astrovirus (GoAstV), a recently identified member of the Astroviridae family in China, predominantly affects goslings, resulting in substantial economic losses to the goose farming industry due to its high infection and mortality rates. Currently, the infection mechanism and pathogenesis of GoAstV remain unknown.
Given this, the Viral Overlay Protein Blot Assay was utilized to identify and characterize proteins on the LMH (Leghorn Male Hepatoma) cell membrane that interact with Goose Astrovirus. The identities of the candidate proteins were determined via LC-MS mass spectrometry analysis, bioinformatics analysis, and UniProt database search. The interaction between HSPA5 and the astrovirus protein was further validated in vitro through Western blot and Coimmunoprecipitation experiments. Finally, bioinformatics tools such as SWISSMODEL, AlphaFold, and ZDOCK were employed to construct and analyze the docking complex model between the candidate protein and GoAstV protein, including their key binding residue sites.
We successfully identified a 70 kDa protein in the plasma membrane protein extracts of LMH cells and confirmed the identity of this candidate protein as HSPA5. Meanwhile, experiments further validated the interaction between HSPA5 and astrovirus proteins. Subsequently, we successfully predicted the docking complex model of HSPA5 protein with GoAstV protein. Further prediction of the binding residue sites revealed that seven residues of the GoAstV-P2 protein (THR124, ILE22, VAL24, TRP51, PRO66, GLN100, and VAL125) and twelve residues of the HSPA5 protein (ARG2, HIS3, LEU4, LEU6, ALA7, LEU8, LEU9, LEU10, LEU11, ASP411, VAL413, and LEU415) may be involved in the interaction between these two proteins.
Our research results have preliminarily elucidated the interaction mechanisms between viral proteins and receptors, facilitating exploration from multiple angles of the roles of candidate protein in the process of GoAstV infecting host cells. This provides a theoretical basis for further identification of GoAstV receptors and clarification of its infection mechanisms.
鹅星状病毒（GoAstV）是中国最近在星状病毒科中发现的成员，主要感染雏鹅，因其高感染率和死亡率给养鹅业造成重大经济损失。目前，GoAstV的感染机制和发病机制尚不清楚。
鉴于此，采用病毒覆盖蛋白印迹分析来鉴定和表征与鹅星状病毒相互作用的莱航雄性肝癌（LMH）细胞膜上的蛋白质。通过液相色谱-质谱（LC-MS）分析、生物信息学分析和通用蛋白质数据库搜索来确定候选蛋白质的身份。通过蛋白质免疫印迹和免疫共沉淀实验在体外进一步验证热休克蛋白家族A成员5（HSPA5）与星状病毒蛋白之间的相互作用。最后，使用诸如SWISSMODEL、AlphaFold和ZDOCK等生物信息学工具构建并分析候选蛋白与GoAstV蛋白之间的对接复合物模型，包括它们的关键结合残基位点。
我们在LMH细胞的质膜蛋白提取物中成功鉴定出一种70 kDa的蛋白质，并确认该候选蛋白为HSPA5。同时，实验进一步验证了HSPA5与星状病毒蛋白之间的相互作用。随后，我们成功预测了HSPA5蛋白与GoAstV蛋白的对接复合物模型。对结合残基位点的进一步预测表明，GoAstV-P2蛋白的七个残基（苏氨酸124、异亮氨酸22、缬氨酸24、色氨酸51、脯氨酸66、谷氨酰胺100和缬氨酸125）和HSPA5蛋白的十二个残基（精氨酸2、组氨酸3、亮氨酸4、亮氨酸6、丙氨酸7、亮氨酸8、亮氨酸9、亮氨酸10、亮氨酸11、天冬氨酸411、缬氨酸413和亮氨酸415）可能参与这两种蛋白之间的相互作用。
我们的研究结果初步阐明了病毒蛋白与受体之间的相互作用机制，有助于从多个角度探索候选蛋白在GoAstV感染宿主细胞过程中的作用。这为进一步鉴定GoAstV受体和阐明其感染机制提供了理论依据。
