Sialic acid-containing glycolipids mediate binding and viral entry of SARS-CoV-2

Linh Nguyen; Kelli A McCord; Duong T Bui; Kim M Bouwman; Elena N Kitova; Mohamed Elaish; Dhanraj Kumawat; Gour C Daskhan; Ilhan Tomris; Ling Han; Pradeep Chopra; Tzu-Jing Yang; Steven D Willows; Andrew L Mason; Lara K Mahal; Todd L Lowary; Lori J West; Shang-Te Danny Hsu; Tom Hobman; Stephen M Tompkins; Geert-Jan Boons; Robert P de Vries; Matthew S Macauley; John S Klassen

doi:10.1038/s41589-021-00924-1

Sialic acid-containing glycolipids mediate binding and viral entry of SARS-CoV-2

Linh Nguyen, Kelli A McCord, Duong T Bui, Kim M Bouwman, Elena N Kitova, Mohamed Elaish, Dhanraj Kumawat, Gour C Daskhan, Ilhan Tomris, Ling Han, Pradeep Chopra, Tzu-Jing Yang, Steven D Willows, Andrew L Mason, Lara K Mahal, Todd L Lowary, Lori J West, Shang-Te Danny Hsu, Tom Hobman, Stephen M TompkinsGeert-Jan Boons, Robert P de Vries, Matthew S Macauley, John S Klassen

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

Abstract

Emerging evidence suggests that host glycans influence severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Here, we reveal that the receptor-binding domain (RBD) of the spike (S) protein on SARS-CoV-2 recognizes oligosaccharides containing sialic acid (Sia), with preference for monosialylated gangliosides. Gangliosides embedded within an artificial membrane also bind to the RBD. The monomeric affinities (K_d = 100–200 μM) of gangliosides for the RBD are similar to another negatively charged glycan ligand of the RBD proposed as a viral co-receptor, heparan sulfate (HS) dp2–dp6 oligosaccharides. RBD binding and infection of SARS-CoV-2 pseudotyped lentivirus to angiotensin-converting enzyme 2 (ACE2)-expressing cells is decreased following depletion of cell surface Sia levels using three approaches: sialyltransferase (ST) inhibition, genetic knockout of Sia biosynthesis, or neuraminidase treatment. These effects on RBD binding and both pseudotyped and authentic SARS-CoV-2 viral entry are recapitulated with pharmacological or genetic disruption of glycolipid biosynthesis. Together, these results suggest that sialylated glycans, specifically glycolipids, facilitate viral entry of SARS-CoV-2. [Figure not available: see fulltext.]

Original language	English
Pages (from-to)	81-90
Number of pages	10
Journal	Nature Chemical Biology
Volume	18
Issue number	1
DOIs	https://doi.org/10.1038/s41589-021-00924-1
Publication status	Published - 9 Nov 2021

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s41589-021-00924-1Final published version, 5.66 MBLicence: Taverne

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Nguyen, L., McCord, K. A., Bui, D. T., Bouwman, K. M., Kitova, E. N., Elaish, M., Kumawat, D., Daskhan, G. C., Tomris, I., Han, L., Chopra, P., Yang, T.-J., Willows, S. D., Mason, A. L., Mahal, L. K., Lowary, T. L., West, L. J., Hsu, S.-T. D., Hobman, T., ... Klassen, J. S. (2021). Sialic acid-containing glycolipids mediate binding and viral entry of SARS-CoV-2. Nature Chemical Biology, 18(1), 81-90. https://doi.org/10.1038/s41589-021-00924-1

@article{13c4066dc59e481592369aa9306c742f,

title = "Sialic acid-containing glycolipids mediate binding and viral entry of SARS-CoV-2",

abstract = "Emerging evidence suggests that host glycans influence severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Here, we reveal that the receptor-binding domain (RBD) of the spike (S) protein on SARS-CoV-2 recognizes oligosaccharides containing sialic acid (Sia), with preference for monosialylated gangliosides. Gangliosides embedded within an artificial membrane also bind to the RBD. The monomeric affinities (Kd = 100–200 μM) of gangliosides for the RBD are similar to another negatively charged glycan ligand of the RBD proposed as a viral co-receptor, heparan sulfate (HS) dp2–dp6 oligosaccharides. RBD binding and infection of SARS-CoV-2 pseudotyped lentivirus to angiotensin-converting enzyme 2 (ACE2)-expressing cells is decreased following depletion of cell surface Sia levels using three approaches: sialyltransferase (ST) inhibition, genetic knockout of Sia biosynthesis, or neuraminidase treatment. These effects on RBD binding and both pseudotyped and authentic SARS-CoV-2 viral entry are recapitulated with pharmacological or genetic disruption of glycolipid biosynthesis. Together, these results suggest that sialylated glycans, specifically glycolipids, facilitate viral entry of SARS-CoV-2. [Figure not available: see fulltext.]",

author = "Linh Nguyen and McCord, {Kelli A} and Bui, {Duong T} and Bouwman, {Kim M} and Kitova, {Elena N} and Mohamed Elaish and Dhanraj Kumawat and Daskhan, {Gour C} and Ilhan Tomris and Ling Han and Pradeep Chopra and Tzu-Jing Yang and Willows, {Steven D} and Mason, {Andrew L} and Mahal, {Lara K} and Lowary, {Todd L} and West, {Lori J} and Hsu, {Shang-Te Danny} and Tom Hobman and Tompkins, {Stephen M} and Geert-Jan Boons and {de Vries}, {Robert P} and Macauley, {Matthew S} and Klassen, {John S}",

note = "Funding Information: We acknowledge the Natural Sciences and Engineering Research Council of Canada (RGPIN-2019-06771 to J.S.K., RGPIN-2018-03815 to M.S.M.), ERC Starting Grant from the European Commission (802780 to R.P.dV), NIH National Heart, Lung and Blood Institute (R01HL151617 to G.-J.B.) the Canadian Glycomics Network, the Canada Foundation for Innovation and the Alberta Innovation and Advanced Education Research Capacity Program and the Beijerinck Premium of the Royal Dutch Academy of Sciences to R.P.dV. for generous funding. L.K.M. thanks the Canada Excellence Research Chair Program for funding. M.S.M. and L.J.W. thank Canada Research Chairs for a Chair in Chemical Glycoimmunology and Cardiac Transplantation, respectively. We thank D. J. Marchant (University of Alberta) for advice and critical reading the manuscript, J. Nagendran (University of Alberta) for generously providing the lung tissue, K. Susuki (University of Alberta) for preparation of lung homogenate, P. D. Bieniasz (The Rockefeller University) and J. L. M. Law (University of Alberta) for providing reagents (plasmids) for generating the pseudotyped virus and ACE2+ HEK293 cells, C. Cairo (University of Alberta) for the endoglycoceramidase enzyme and S. Sipione (University of Alberta) for advice related to glycolipids. mPlum-ER-3 was a gift from M. Davidson (Addgene plasmid 55966; http:// n2t.net/addgene:55966; RRID:Addgene_55966). Plasmids for expression of SARS-CoV-2 S protein and RBD proteins were generously provided by F. Krammer (Icahn School of Medicine at Mount Sinai, produced under NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS) contract HHSN272201400008C) and produced under CEIRS contract HHSN272201400004C awarded to S.M.T. SARS-CoV-2 VIDO strain (hCoV-19/Canada/ON-VIDO-01/2020, GISAID accession EPI_ISL_425177) was kindly provided by D. Falzarano (Vaccine and Infectious Disease Organization). The SARS-CoV-2 72b Calg strain (72B/CA/CALG) was kindly provided by D. Evans (University of Alberta). Publisher Copyright: {\textcopyright} 2021, The Author(s), under exclusive licence to Springer Nature America, Inc.",

year = "2021",

month = nov,

day = "9",

doi = "10.1038/s41589-021-00924-1",

language = "English",

volume = "18",

pages = "81--90",

journal = "Nature Chemical Biology",

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Nguyen, L, McCord, KA, Bui, DT, Bouwman, KM, Kitova, EN, Elaish, M, Kumawat, D, Daskhan, GC, Tomris, I, Han, L, Chopra, P, Yang, T-J, Willows, SD, Mason, AL, Mahal, LK, Lowary, TL, West, LJ, Hsu, S-TD, Hobman, T, Tompkins, SM, Boons, G-J , de Vries, RP, Macauley, MS & Klassen, JS 2021, 'Sialic acid-containing glycolipids mediate binding and viral entry of SARS-CoV-2', Nature Chemical Biology, vol. 18, no. 1, pp. 81-90. https://doi.org/10.1038/s41589-021-00924-1

TY - JOUR

T1 - Sialic acid-containing glycolipids mediate binding and viral entry of SARS-CoV-2

AU - Nguyen, Linh

AU - McCord, Kelli A

AU - Bui, Duong T

AU - Bouwman, Kim M

AU - Kitova, Elena N

AU - Elaish, Mohamed

AU - Kumawat, Dhanraj

AU - Daskhan, Gour C

AU - Tomris, Ilhan

AU - Han, Ling

AU - Chopra, Pradeep

AU - Yang, Tzu-Jing

AU - Willows, Steven D

AU - Mason, Andrew L

AU - Mahal, Lara K

AU - Lowary, Todd L

AU - West, Lori J

AU - Hsu, Shang-Te Danny

AU - Hobman, Tom

AU - Tompkins, Stephen M

AU - Boons, Geert-Jan

AU - de Vries, Robert P

AU - Macauley, Matthew S

AU - Klassen, John S

N1 - Funding Information: We acknowledge the Natural Sciences and Engineering Research Council of Canada (RGPIN-2019-06771 to J.S.K., RGPIN-2018-03815 to M.S.M.), ERC Starting Grant from the European Commission (802780 to R.P.dV), NIH National Heart, Lung and Blood Institute (R01HL151617 to G.-J.B.) the Canadian Glycomics Network, the Canada Foundation for Innovation and the Alberta Innovation and Advanced Education Research Capacity Program and the Beijerinck Premium of the Royal Dutch Academy of Sciences to R.P.dV. for generous funding. L.K.M. thanks the Canada Excellence Research Chair Program for funding. M.S.M. and L.J.W. thank Canada Research Chairs for a Chair in Chemical Glycoimmunology and Cardiac Transplantation, respectively. We thank D. J. Marchant (University of Alberta) for advice and critical reading the manuscript, J. Nagendran (University of Alberta) for generously providing the lung tissue, K. Susuki (University of Alberta) for preparation of lung homogenate, P. D. Bieniasz (The Rockefeller University) and J. L. M. Law (University of Alberta) for providing reagents (plasmids) for generating the pseudotyped virus and ACE2+ HEK293 cells, C. Cairo (University of Alberta) for the endoglycoceramidase enzyme and S. Sipione (University of Alberta) for advice related to glycolipids. mPlum-ER-3 was a gift from M. Davidson (Addgene plasmid 55966; http:// n2t.net/addgene:55966; RRID:Addgene_55966). Plasmids for expression of SARS-CoV-2 S protein and RBD proteins were generously provided by F. Krammer (Icahn School of Medicine at Mount Sinai, produced under NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS) contract HHSN272201400008C) and produced under CEIRS contract HHSN272201400004C awarded to S.M.T. SARS-CoV-2 VIDO strain (hCoV-19/Canada/ON-VIDO-01/2020, GISAID accession EPI_ISL_425177) was kindly provided by D. Falzarano (Vaccine and Infectious Disease Organization). The SARS-CoV-2 72b Calg strain (72B/CA/CALG) was kindly provided by D. Evans (University of Alberta). Publisher Copyright: © 2021, The Author(s), under exclusive licence to Springer Nature America, Inc.

PY - 2021/11/9

Y1 - 2021/11/9

N2 - Emerging evidence suggests that host glycans influence severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Here, we reveal that the receptor-binding domain (RBD) of the spike (S) protein on SARS-CoV-2 recognizes oligosaccharides containing sialic acid (Sia), with preference for monosialylated gangliosides. Gangliosides embedded within an artificial membrane also bind to the RBD. The monomeric affinities (Kd = 100–200 μM) of gangliosides for the RBD are similar to another negatively charged glycan ligand of the RBD proposed as a viral co-receptor, heparan sulfate (HS) dp2–dp6 oligosaccharides. RBD binding and infection of SARS-CoV-2 pseudotyped lentivirus to angiotensin-converting enzyme 2 (ACE2)-expressing cells is decreased following depletion of cell surface Sia levels using three approaches: sialyltransferase (ST) inhibition, genetic knockout of Sia biosynthesis, or neuraminidase treatment. These effects on RBD binding and both pseudotyped and authentic SARS-CoV-2 viral entry are recapitulated with pharmacological or genetic disruption of glycolipid biosynthesis. Together, these results suggest that sialylated glycans, specifically glycolipids, facilitate viral entry of SARS-CoV-2. [Figure not available: see fulltext.]

AB - Emerging evidence suggests that host glycans influence severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Here, we reveal that the receptor-binding domain (RBD) of the spike (S) protein on SARS-CoV-2 recognizes oligosaccharides containing sialic acid (Sia), with preference for monosialylated gangliosides. Gangliosides embedded within an artificial membrane also bind to the RBD. The monomeric affinities (Kd = 100–200 μM) of gangliosides for the RBD are similar to another negatively charged glycan ligand of the RBD proposed as a viral co-receptor, heparan sulfate (HS) dp2–dp6 oligosaccharides. RBD binding and infection of SARS-CoV-2 pseudotyped lentivirus to angiotensin-converting enzyme 2 (ACE2)-expressing cells is decreased following depletion of cell surface Sia levels using three approaches: sialyltransferase (ST) inhibition, genetic knockout of Sia biosynthesis, or neuraminidase treatment. These effects on RBD binding and both pseudotyped and authentic SARS-CoV-2 viral entry are recapitulated with pharmacological or genetic disruption of glycolipid biosynthesis. Together, these results suggest that sialylated glycans, specifically glycolipids, facilitate viral entry of SARS-CoV-2. [Figure not available: see fulltext.]

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U2 - 10.1038/s41589-021-00924-1

DO - 10.1038/s41589-021-00924-1

M3 - Article

C2 - 34754101

SN - 1552-4450

VL - 18

SP - 81

EP - 90

JO - Nature Chemical Biology

JF - Nature Chemical Biology

IS - 1

ER -

Sialic acid-containing glycolipids mediate binding and viral entry of SARS-CoV-2

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