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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 #334186

Research Project: Ecology and Detection of Human Pathogens in the Produce Production Continuum

Location: Produce Safety and Microbiology Research

Title: Bacterial surface-displayed GII.4 human norovirus capsid proteins bound to HBGA-like molecules in romaine lettuce

Author
item Wang, Ming - Shanghai Institute Of Technology
item Rong, Shaofeng - Shanghai Institute Of Technology
item Tian, Peng
item Zhou, Yue - Shanghai Institute Of Technology
item Guan, Shimin - Shanghai Institute Of Technology
item Li, Quianqian - Shanghai Institute Of Technology
item Wang, Dapeng - Shanghai Institute Of Technology

Submitted to: Frontiers in Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/6/2017
Publication Date: 2/20/2017
Citation: Wang, M., Rong, S., Tian, P., Zhou, Y., Guan, S., Li, Q., Wang, D. 2017. Bacterial surface-displayed GII.4 human norovirus capsid proteins bound to HBGA-like molecules in romaine lettuce. Frontiers in Microbiology. 8:251. https://doi.org/10.3389/fmicb.2017.00251.
DOI: https://doi.org/10.3389/fmicb.2017.00251

Interpretive Summary: Human noroviruses are highly-contagious pathogens and is the main cause of acute gastroenteritis. GII.4 genotype of human norovirus is the dominant strain causing acute gastroenteritis worldwide. In recent years, we have seen an increase in the number of HuNoV outbreaks, which correlates with the increased consumption of minimally-processed fruits and vegetables. In this study, we used an ice nucleation protein (INP) mediated surface display system to present GII.4 HuNoV capsid protein on bacterial surface and used it as a new strategy to explore interaction between the displayed HuNoV capsid P protein and candidate receptors from Romaine lettuce. The surface displayed GII.4 P proteins were confirmed on the surface of the transformed bacteria by an immunofluorescence assay (IF). The distribution patterns of HuNoV P proteins on the surface of Romaine lettuce were identified through a confocal IF assay. HuNoV P proteins could be found around stoma and surface of vein and leaf of Romaine lettuce. HuNoV P proteins could be captured by extract from leaf (LE) or vein (EV) in an ELISA assay. The binding of HuNoV P proteins to LE or VE could be competitively blocked by histo-blood group antigens (HBGAs) from human saliva. In addition, the binding of HuNoV P proteins to LE or VE could also be completely demolished when carbohydrates in LE or VE were oxidized. The binding of HuNoV P proteins to LE or VE could only be partially inhibited by heat inactivation of lettuce proteins. The results indicated that HBGA-like glycoproteins in LE or VE were likely contributed to binding of the HuNoV P proteins to Romaine lettuce. We demonstrated that the surface displayed HuNoV P proteins could be a new and simple system for investigation the interaction between the HuNoVs and their candidate receptors.

Technical Abstract: Human Noroviruses (HuNoVs) are the main cause of nonbacterial gastroenteritis. Contaminated produce is a main vehicle for dissemination of HuNoVs. In this study, we used an ice nucleation protein (INP) mediated surface display system to present the protruding domain of GII.4 HuNoV capsid protein (GII.4 P) on bacterial surface and used it as a new strategy to explore interaction between the surface-displayed HuNoV capsid P protein (SD-GII.4 P) and candidate receptors from Romaine lettuce. SD-GII.4 P from INP-P (GII.4) gene transformed E. coli BL 21 bacteria [INP-P (GII.4) BL21] were confirmed on the surface of the transformed bacteria by an immunofluorescence assay (IF). The distribution patterns of SD-GII.4 P on the surface of Romaine lettuce were identified through a confocal IF assay. SD-GII.4 P could be found around stoma and surface of vein and leaf of Romaine lettuce. SD-GII.4 P could be captured by extract from leaf (LE) or vein (EV) in an ELISA assay. The binding of SD-GII.4 P to LE or VE could be competitively blocked by histo-blood group antigens (HBGAs) from human saliva. In addition, the binding of SD-GII.4 P to LE or VE could also be completely demolished when LE or VE were oxidized by sodium periodate and partially inhibited by heat inactivation of lettuce proteins. The results indicated that HBGA-like glycoproteins in LE or VE were likely contributed to binding of SD-GII.4 P to Romaine lettuce. We demonstrated that SD-GII.4 P could be a new and simple system for investigation the interaction between the HuNoVs and their candidate receptors.