SARS-CoV-2 Infection Depends on Cellular Heparan Sulfate and ACE2

Thomas Mandel Clausen; Daniel R Sandoval; Charlotte B Spliid; Jessica Pihl; Hailee R Perrett; Chelsea D Painter; Anoop Narayanan; Sydney A Majowicz; Elizabeth M Kwong; Rachael N McVicar; Bryan E Thacker; Charles A Glass; Zhang Yang; Jonathan L Torres; Gregory J Golden; Phillip L Bartels; Ryan N Porell; Aaron F Garretson; Logan Laubach; Jared Feldman; Xin Yin; Yuan Pu; Blake M Hauser; Timothy M Caradonna; Benjamin P Kellman; Cameron Martino; Philip L S M Gordts; Sumit K Chanda; Aaron G Schmidt; Kamil Godula; Sandra L Leibel; Joyce Jose; Kevin D Corbett; Andrew B Ward; Aaron F Carlin; Jeffrey D Esko

doi:10.1016/j.cell.2020.09.033

SARS-CoV-2 Infection Depends on Cellular Heparan Sulfate and ACE2

Cell. 2020 Nov 12;183(4):1043-1057.e15. doi: 10.1016/j.cell.2020.09.033. Epub 2020 Sep 14.

Authors

Thomas Mandel Clausen¹, Daniel R Sandoval², Charlotte B Spliid³, Jessica Pihl³, Hailee R Perrett⁴, Chelsea D Painter⁵, Anoop Narayanan⁶, Sydney A Majowicz⁶, Elizabeth M Kwong⁷, Rachael N McVicar⁷, Bryan E Thacker⁸, Charles A Glass⁸, Zhang Yang⁹, Jonathan L Torres⁴, Gregory J Golden⁵, Phillip L Bartels¹⁰, Ryan N Porell¹¹, Aaron F Garretson¹², Logan Laubach¹¹, Jared Feldman¹³, Xin Yin¹⁴, Yuan Pu¹⁴, Blake M Hauser¹³, Timothy M Caradonna¹³, Benjamin P Kellman¹⁵, Cameron Martino¹⁶, Philip L S M Gordts¹⁷, Sumit K Chanda¹⁴, Aaron G Schmidt¹⁸, Kamil Godula¹⁹, Sandra L Leibel²⁰, Joyce Jose⁶, Kevin D Corbett², Andrew B Ward⁴, Aaron F Carlin¹², Jeffrey D Esko²¹

Affiliations

¹ Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA; Department for Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark; Department of Infectious Disease, Copenhagen University Hospital, 2200 Copenhagen, Denmark. Electronic address: tmandelclausen@health.ucsd.edu.
² Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA.
³ Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA; Department for Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark; Department of Infectious Disease, Copenhagen University Hospital, 2200 Copenhagen, Denmark.
⁴ Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
⁵ Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA; Biomedical Sciences Graduate Program, University of California San Diego, La Jolla, California, USA.
⁶ Department of Biochemistry and Molecular Biology, The Huck Institutes of the Life Sciences, Pennsylvania State University, University Park, PA 16802, USA.
⁷ Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA 92037, USA.
⁸ TEGA Therapeutics, Inc., 3550 General Atomics Court, G02-102, San Diego, CA 92121, USA.
⁹ Copenhagen Center for Glycomics, Department of Molecular and Cellular Medicine, University of Copenhagen, 2200 Copenhagen, Denmark.
¹⁰ Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA; Glycobiology Research and Training Center, University of California, San Diego, La Jolla, CA 92093, USA.
¹¹ Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093, USA.
¹² Department of Medicine, University of California, San Diego, La Jolla, CA 92037, USA.
¹³ Ragon Institute of MGH, MIT and Harvard, Cambridge, MA 02139, USA.
¹⁴ Immunity and Pathogenesis Program, Infectious and Inflammatory Disease Center, Sanford Burnham Prebys Medical Discovery Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, USA.
¹⁵ Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093, USA; Department of Pediatrics, University of California San Diego School of Medicine, La Jolla, CA 92093, USA; Bioinformatics and Systems Biology Program, University of California, San Diego, La Jolla, CA 92093, USA.
¹⁶ Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093, USA; Department of Pediatrics, University of California San Diego School of Medicine, La Jolla, CA 92093, USA.
¹⁷ Department of Medicine, University of California, San Diego, La Jolla, CA 92037, USA; Glycobiology Research and Training Center, University of California, San Diego, La Jolla, CA 92093, USA.
¹⁸ Ragon Institute of MGH, MIT and Harvard, Cambridge, MA 02139, USA; Department of Microbiology, Harvard Medical School, Boston, MA 02115, USA.
¹⁹ Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093, USA; Glycobiology Research and Training Center, University of California, San Diego, La Jolla, CA 92093, USA.
²⁰ Department of Pediatrics, University of California San Diego School of Medicine, La Jolla, CA 92093, USA.
²¹ Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA; Glycobiology Research and Training Center, University of California, San Diego, La Jolla, CA 92093, USA. Electronic address: jesko@health.ucsd.edu.

Abstract

We show that SARS-CoV-2 spike protein interacts with both cellular heparan sulfate and angiotensin-converting enzyme 2 (ACE2) through its receptor-binding domain (RBD). Docking studies suggest a heparin/heparan sulfate-binding site adjacent to the ACE2-binding site. Both ACE2 and heparin can bind independently to spike protein in vitro, and a ternary complex can be generated using heparin as a scaffold. Electron micrographs of spike protein suggests that heparin enhances the open conformation of the RBD that binds ACE2. On cells, spike protein binding depends on both heparan sulfate and ACE2. Unfractionated heparin, non-anticoagulant heparin, heparin lyases, and lung heparan sulfate potently block spike protein binding and/or infection by pseudotyped virus and authentic SARS-CoV-2 virus. We suggest a model in which viral attachment and infection involves heparan sulfate-dependent enhancement of binding to ACE2. Manipulation of heparan sulfate or inhibition of viral adhesion by exogenous heparin presents new therapeutic opportunities.

Keywords: COVID-19; SARS-CoV-2; coronavirus; heparan sulfate; heparan sulfate-binding proteins; heparin; lung epithelial cells; pseudotyped virus; spike proteins.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Angiotensin-Converting Enzyme 2
Betacoronavirus / isolation & purification
Betacoronavirus / physiology*
Binding Sites
COVID-19
Cell Line
Coronavirus Infections / pathology
Coronavirus Infections / virology
Heparin / chemistry
Heparin / metabolism
Heparitin Sulfate / chemistry
Heparitin Sulfate / metabolism*
Humans
Kidney / metabolism
Lung / metabolism
Molecular Dynamics Simulation
Pandemics
Peptidyl-Dipeptidase A / chemistry
Peptidyl-Dipeptidase A / metabolism*
Pneumonia, Viral / pathology
Pneumonia, Viral / virology
Protein Binding
Protein Domains
Recombinant Proteins / biosynthesis
Recombinant Proteins / chemistry
Recombinant Proteins / isolation & purification
SARS-CoV-2
Spike Glycoprotein, Coronavirus / chemistry
Spike Glycoprotein, Coronavirus / genetics
Spike Glycoprotein, Coronavirus / metabolism*
Virus Internalization

Substances

Recombinant Proteins
Spike Glycoprotein, Coronavirus
spike protein, SARS-CoV-2
Heparin
Heparitin Sulfate
Peptidyl-Dipeptidase A
ACE2 protein, human
Angiotensin-Converting Enzyme 2

Abstract

Publication types

MeSH terms

Substances

Grants and funding