Comprehensive characterization of N- and O- glycosylation of SARS-CoV-2 human receptor angiotensin converting enzyme 2

Shajahan, Asif; Archer-Hartmann, Stephanie; Supekar, Nitin T.; Gleinich, Anne S.; Heiss, Christian; Azadi, Parastoo

doi:10.1093/glycob/cwaa101

Comprehensive characterization of N- and O- glycosylation of SARS-CoV-2 human receptor angiotensin converting enzyme 2

Journal Article · Thu Oct 29 00:00:00 EDT 2020 · Glycobiology (Online)

DOI:https://doi.org/10.1093/glycob/cwaa101· OSTI ID:1852017

^[1]; Archer-Hartmann, Stephanie ^[2]; Supekar, Nitin T. ^[2]; ^[2]; Heiss, Christian ^[2]; Azadi, Parastoo ^[2]

Complex Carbohydrate Research Center, The University of Georgia, 315 Riverbend Rd, Athens, GA 30602, USA; OSTI
Complex Carbohydrate Research Center, The University of Georgia, 315 Riverbend Rd, Athens, GA 30602, USA

Abstract

The emergence of the coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has created the need for development of new therapeutic strategies. Understanding the mode of viral attachment, entry and replication has become a key aspect of such interventions. The coronavirus surface features a trimeric spike (S) protein that is essential for viral attachment, entry and membrane fusion. The S protein of SARS-CoV-2 binds to human angiotensin converting enzyme 2 (hACE2) for entry. Herein, we describe glycomic and glycoproteomic analysis of hACE2 expressed in HEK293 cells. We observed high glycan occupancy (73.2 to 100%) at all seven possible N-glycosylation sites and surprisingly detected one novel O-glycosylation site. To deduce the detailed structure of glycan epitopes on hACE2 that may be involved in viral binding, we have characterized the terminal sialic acid linkages, the presence of bisecting GlcNAc and the pattern of N-glycan fucosylation. We have conducted extensive manual interpretation of each glycopeptide and glycan spectrum, in addition to using bioinformatics tools to validate the hACE2 glycosylation. Our elucidation of the site-specific glycosylation and its terminal orientations on the hACE2 receptor, along with the modeling of hACE2 glycosylation sites can aid in understanding the intriguing virus-receptor interactions and assist in the development of novel therapeutics to prevent viral entry. The relevance of studying the role of ACE2 is further increased due to some recent reports about the varying ACE2 dependent complications with regard to age, sex, race and pre-existing conditions of COVID-19 patients.

Research Organization:: Univ. of Georgia, Athens, GA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

DOE Contract Number:: SC0015662

OSTI ID:: 1852017

Journal Information:: Glycobiology (Online), Journal Name: Glycobiology (Online) Journal Issue: 4 Vol. 31; ISSN 1460-2423

Publisher:: Oxford University Press

Country of Publication:: United States

Language:: English

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