Screening and Identification of Host Proteins Interacting with PDCoV S1-CTD
Acta Veterinaria et Zootechnica Sinica
; 53(7):2260-2267, 2022.
Article
in Chinese
| EMBASE | ID: covidwho-2245652
ABSTRACT
The C-terminal domain (CTD) of porcine deltacoronavirus S1 subunit is the main region which induces the neutralizing antibody. S1-CTD was expressed by HEK-293T eukaryotic expression system and purified, and porcine ileal epithelium cells membrane proteins were extracted to investigate porcine host proteins that interact with it. Thirty-two suspected interacting host proteins were obtained by co-inmunprecipitation (Co-IP) and mass spectrometry. Eukaryotic expression plasmid of KIF1 binding protein (KIFBP) was constructed, and the interaction between KIFBP and S1-CTD was identified by Co-IP and laser confocal microscopy. All results proved that KIFBP interacted with S1-CTD and co-located in cytoplasm. Further research indicated that overexpression of KIFBP could effectively reduce the viral mRNA level and the viral titer in which the mRNA level decreased by about 70%, and the viral titer decreased by 101.6TCID50. In conclusion, a host protein KIFBP interacting with PDCoV S1-CTD was screened and identified in this study which provides a theoretical basis for understanding the pathogenesis of PDCoV.
binding protein; KIF1 binding protein; unclassified drug; virus messenger RNA; article; carboxy terminal sequence; coimmunoprecipitation; confocal laser scanning microscopy; cytoplasm; eukaryote; HEK293T cell line; human; human cell; mass spectrometry; plasmid; porcine deltacoronavirus infection; protein analysis; protein expression; protein interaction; TCID50; theoretical study
Full text:
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Collection:
Databases of international organizations
Database:
EMBASE
Language:
Chinese
Journal:
Acta Veterinaria et Zootechnica Sinica
Year:
2022
Document Type:
Article
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