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ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Produce Safety and Microbiology Research » Research » Publications at this Location » Publication #320742
Title: A Novel system for evaluating the interaction between human norovirus and receptors

Author
item Tian, Peng
item WANG, DAPENG - University Of Shanghai
item NIU, MENGYA - University Of Shanghai
item YU, QIANQIAN - University Of Shanghai
item GAO, ZHIYONG - University Of Shanghai
item SHI, XIANMING - University Of Shanghai

Submitted to: Frontiers in Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/30/2015
Publication Date: N/A
Citation: N/A

Interpretive Summary: Human noroviruses (HuNoV) are pathogens of significant concern, but as of current they cannot be cultured, which limits the ways in which they can be studied. Scientists at Produce Safety and Microbiology Research Unit at WRRC and MOST-USDA Joint Research Center for Food Safety, School of Agriculture & Biology, Shanghai Jiao Tong University, Shanghai, China engineered a novel system to express the viral capsid proteins on the surface of a bacteria. The new system could also be used for isolation unknown receptors for human noroviruses and exploring interaction between human norovirus and receptors. By transforming a bacteria with the gene encoding a fusion target protein with the membrane anchoring motifs, the target proteins could be directly displayed on the surface of the bacteria. We demonstrated that the surface displayed viral proteins could effectively bind to the viral receptor. More importantly, the viral protein-ligand complex could be easily isolated by a simple low speed centrifugation. Therefore, the strategy of using bacterial surface displayed HuNoV capsid proteins could be useful to separate unknown human norovirus binding components from complex samples and to investigate interaction between the virus and its receptors. In addition, the new system could be a good candidate for vaccine development.

Technical Abstract: Human noroviruses (HuNoVs) are major pathogens for acute nonbacterial gastroenteritis outbreaks. Many aspects of HuNoVs are poorly understood due to both the current inability to culture HuNoVs, and the lack of efficient small animal models. Recombinant HuNoV viral capsid proteins and/or P particles have been used as surrogates for HuNoV. In this study, we used the ice nucleation protein to display capsid proteins of HuNoV on bacterial surface as a new strategy to explore interaction between capsid proteins of HuNoV and its ligand. The VP1 capsid encoding gene (ORF2) and P particle encoding gene (protruding domain) of HuNoV GI.1 and GII.4 were fused with ice nucleation protein coding gene (InaQ). We demonstrated that the VP1s and P particles of HuNoV were expressed and anchored on the surface of E.coli BL21 cells after the bacteria was transformed with the corresponding plasmids. The cell surface displayed HuNoV VP1s and P particles could be recognized by HuNoV specific antibodies. The displayed VP1 and P particles also could interact with the viral histo-blood group antigens receptors. The viral capsid protein-ligand complex could be isolated easily by a simple low speed centrifugation. Therefore, the strategy of using bacterial surface displayed HuNoV capsid proteins could be particularly useful to separate unknown HuNoV binding components from complex samples and to investigate interaction between the virus and its receptors. In addition, the new system could be a good candidate for vaccine development.