Bacterial glycosyltransferase-mediated cell-surface chemoenzymatic glycan modification

Senlian Hong; Yujie Shi; Nicholas C. Wu; Geramie Grande; Lacey Douthit; Hua Wang; Wen Zhou; K. Barry Sharpless; Ian A. Wilson; Jia Xie; Peng Wu

doi:10.1038/s41467-019-09608-w

Bacterial glycosyltransferase-mediated cell-surface chemoenzymatic glycan modification

Senlian Hong, Yujie Shi, Nicholas C. Wu, Geramie Grande, Lacey Douthit, Hua Wang, Wen Zhou, K. Barry Sharpless, Ian A. Wilson, Jia Xie, Peng Wu

Research output: Contribution to journal › Article › peer-review

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 (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.

Original language	English (US)
Article number	1799
Journal	Nature communications
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41467-019-09608-w
State	Published - Dec 1 2019
Externally published	Yes

ASJC Scopus subject areas

Chemistry(all)
Biochemistry, Genetics and Molecular Biology(all)
Physics and Astronomy(all)

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10.1038/s41467-019-09608-w

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title = "Bacterial glycosyltransferase-mediated cell-surface chemoenzymatic glycan modification",

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 (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. ",

author = "Senlian Hong and Yujie Shi and Wu, {Nicholas C.} and Geramie Grande and Lacey Douthit and Hua Wang and Wen Zhou and Sharpless, {K. Barry} and Wilson, {Ian A.} and Jia Xie and Peng Wu",

note = "Funding Information: We thank Prof. Richard A. Lerner (The Scripps Research Institute), Prof. J. C. Paulson (The Scripps Research Institute), Prof. Kay-Hooi Khoo (Institute of Biological Chemistry, Academia Sinica, Taiwan), and Prof. Carlito B. Lebrilla (Department of Chemistry and Biochemistry, University of California, Davis, USA) for insightful discussions. This work was supported by Grants from the NIH (P.W. GM093282 and GM113046, K.B.S. P50 GM103368, I.A.W. R56 AI127371 and BMGF OPP1170236). Publisher Copyright: {\textcopyright} 2019, This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.",

year = "2019",

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doi = "10.1038/s41467-019-09608-w",

language = "English (US)",

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AU - Hong, Senlian

AU - Shi, Yujie

AU - Wu, Nicholas C.

AU - Grande, Geramie

AU - Douthit, Lacey

AU - Wang, Hua

AU - Zhou, Wen

AU - Sharpless, K. Barry

AU - Wilson, Ian A.

AU - Xie, Jia

AU - Wu, Peng

N1 - Funding Information: We thank Prof. Richard A. Lerner (The Scripps Research Institute), Prof. J. C. Paulson (The Scripps Research Institute), Prof. Kay-Hooi Khoo (Institute of Biological Chemistry, Academia Sinica, Taiwan), and Prof. Carlito B. Lebrilla (Department of Chemistry and Biochemistry, University of California, Davis, USA) for insightful discussions. This work was supported by Grants from the NIH (P.W. GM093282 and GM113046, K.B.S. P50 GM103368, I.A.W. R56 AI127371 and BMGF OPP1170236). Publisher Copyright: © 2019, This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.

PY - 2019/12/1

Y1 - 2019/12/1

N2 - 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.

AB - 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.

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Bacterial glycosyltransferase-mediated cell-surface chemoenzymatic glycan modification

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