Bacterial glycosyltransferase-mediated cell-surface chemoenzymatic glycan modification

• Published in: Nature communications 2019-04, Vol.10 (1), p.1799-11, Article 1799
• Main Authors: Hong, Senlian, Shi, Yujie, Wu, Nicholas C., Grande, Geramie, Douthit, Lacey, Wang, Hua, Zhou, Wen, Sharpless, K. Barry, Wilson, Ian A., Xie, Jia, Wu, Peng
• Format: Article
• Language: English
• Subjects: 13/1; 13/105; 13/31; 13/51; 14/34; 14/63; 38/109; 38/5; 639/638/92/2783; 639/638/92/507; 639/638/92/72/1205; 64/60; 82/80; 82/83; Animals; Antigens; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biological Assay - methods; Blood groups; Cell death; Cell surface; Chemistry; CHO Cells; Cricetulus; Dogs; Enzymes; Fucose; Glycan; Glycosyltransferase; Glycosyltransferases - genetics; Glycosyltransferases - metabolism; Healthy Volunteers; Helicobacter mustelae - genetics; Helicobacter mustelae - metabolism; Helicobacter pylori; Hemagglutinins; Hemagglutinins - immunology; Hemagglutinins - metabolism; Host-Pathogen Interactions - immunology; Humanities and Social Sciences; Humans; Hydrogen bonds; Hydroxyl groups; Influenza; Influenza A; Influenza A Virus, H1N1 Subtype - genetics; Influenza A Virus, H1N1 Subtype - immunology; Influenza A Virus, H3N2 Subtype - genetics; Influenza A Virus, H3N2 Subtype - immunology; Influenza, Human - immunology; Influenza, Human - virology; Intravital Microscopy - methods; Labeling; Lectins; Ligands; Luciferases, Bacterial - genetics; Luciferases, Bacterial - metabolism; Lung - pathology; Madin Darby Canine Kidney Cells; Metabolic engineering; Metabolic Engineering - methods; Metabolism; Microscopy, Fluorescence - methods; Molecular Imaging - methods; multidisciplinary; Oligosaccharides; Oligosaccharides - immunology; Oligosaccharides - metabolism; Pasteurella multocida - genetics; Pasteurella multocida - metabolism; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Research centers; Science; Science (multidisciplinary); Sialyl Lewis X Antigen; Staining and Labeling - methods; Strains (organisms); Viruses
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-019-09608-w

Chemoenzymatic modification of cell-surface glycan structures has emerged as a complementary approach to metabolic oligosaccharide engineering. Here, we identify Pasteurella multocida α2-3-sialyltransferase M144D mutant, Photobacterium damsela α2-6-sialyltransferase, and Helicobacter mustelae α1-2-fucosyltransferase, as efficient tools for live-cell glycan modification. Combining these enzymes with Helicobacter pylori α1-3-fucosyltransferase, we develop a host-cell-based assay to probe glycan-mediated influenza A virus (IAV) infection including wild-type and mutant strains of H1N1 and H3N2 subtypes. At high NeuAcα2-6-Gal levels, the IAV-induced host-cell death is positively correlated with haemagglutinin (HA) binding affinity to NeuAcα2-6-Gal. Remarkably, an increment of host-cell-surface sialyl Lewis X (sLe X ) exacerbates the killing by several wild-type IAV strains and a previously engineered mutant HK68-MTA. Structural alignment of HAs from HK68 and HK68-MTA suggests formation of a putative hydrogen bond between Trp222 of HA-HK68-MTA and the C-4 hydroxyl group of the α1-3-linked fucose of sLe X , which may account for the enhanced host cell killing of that mutant. Glycan molecules can be modified directly on the cell surface via chemoenzymatic approaches. Here, the authors employ a set of four bacterial glycosyltransferases to develop a live cell-based killing assay to probe host cell glycan-mediated influenza A virus infection.