Bacterial glycosyltransferase-mediated cell-surface chemoenzymatic glycan modification

  • Nat Commun. 2019 Apr 17;10(1):1799. doi: 10.1038/s41467-019-09608-w.
Senlian Hong  1 ,  Yujie Shi  1 ,  Nicholas C Wu  2 ,  Geramie Grande  3 ,  Lacey Douthit  3 ,  Hua Wang  3 ,  Wen Zhou  4 ,  K Barry Sharpless  3 ,  Ian A Wilson  2  5 ,  Jia Xie  6 ,  Peng Wu  7
Affiliations
  • 1. Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA, 92037, USA.
  • 2. Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA, 92037, USA.
  • 3. Department of Chemistry, The Scripps Research Institute, La Jolla, CA, 92037, USA.
  • 4. College of Chemistry and Molecular Engineering, Peking University, 100871, Beijing, China.
  • 5. Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA, 92037, USA.
  • 6. Department of Chemistry, The Scripps Research Institute, La Jolla, CA, 92037, USA. [email protected].
  • 7. Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA, 92037, USA. [email protected].
PMID: 30996301 DOI: 10.1038/s41467-019-09608-w
Abstract

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 (sLeX) 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 sLeX, which may account for the enhanced host cell killing of that mutant.

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