1. Differential white spot syndrome virus-binding proteins in two hemocyte subpopulations of Chinese shrimp (Fenneropenaeus chinensis).
- Author
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Cui, Chuang, Zhu, Lei, Tang, Xiaoqian, Xing, Jing, Sheng, Xiuzhen, Chi, Heng, and Zhan, Wenbin
- Subjects
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BINDING site assay , *WHITE spot syndrome virus , *TRETINOIN , *PROTEIN kinases , *ADENOSINE triphosphatase , *CARRIER proteins , *VIRAL proteins - Abstract
A number of white spot syndrome virus (WSSV)-binding proteins have been identified previously in the hemocytes of Fenneropenaeus chinensis. In order to further investigate the differential WSSV-binding proteins in hemocyte subpopulations, granular hemocytes and hyalinocytes were sorted from WSSV-infected shrimp by immunomagnetic bead (IMB) method. The results of ELISA and immuno-dot blot assay showed that the WSSV-binding activity of granular hemocytes proteins was much stronger than that of hyalinocytes proteins. And the percentage of WSSV-positive granular hemocytes was significantly higher than that of hyalinocytes post WSSV infection, indicating that granular hemocytes were more susceptible to WSSV infection. Moreover, a total of 9 WSSV-binding proteins were successfully identified in granular hemocytes and hyalinocytes by two-dimensional virus overlay protein binding assay (2D-VOPBA) and MALDI-TOF MS analysis, of which 3 binding proteins (arginine kinase, protease 1 and transglutaminase) existing in both hyalinocytes and granular hemocytes and 6 proteins (F 1 ATP synthase β-chain, hnRNPs, GAPDH, RACK1, β-actin and cellular retinoic acid) detected only in granular hemocytes. Among these identified WSSV-binding proteins, the transglutaminase (TG) was further recombinantly expressed, and the recombinant TG could be bound with WSSV. Subsequently, quantitative real-time PCR analysis showed that differential expression levels of WSSV-binding proteins were observed in granular hemocytes and hyalinocytes. The results of this study revealed that the WSSV-binding proteins were differentially expressed in granular hemocytes and hyalinocytes, which provided a deeper insight into the interaction between WSSV and hemocyte subpopulations. • Granulocytes proteins exhibited higher WSSV-binding activity than that of hyalinocytes. • A total of 9 WSSV-binding proteins were identified in granulocytes and hyalinocytes. • 3 WSSV-binding proteins were identified in both granulocytes and hyalinocytes. • 6 WSSV-binding proteins were detected in only granulocytes. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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