摘要：SummaryThe densely glycosylated spike (S) protein highly exposed on severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) surface mediates host cell entry by binding to the receptor angiotensin-converting enzyme 2 (ACE2). However, the role of glycosylation has not been fully understood. In this study, we investigated the effect of differentN-glycosylation of S1 protein on its binding to ACE2. Using real-time surface plasmon resonance assay the negative effects were demonstrated by the considerable increase of binding affinities of de-N-glycosylated S1 proteins produced from three different expression systems including baculovirus-insect, Chinese hamster ovarian and two variants of human embryonic kidney 293 cells. Molecular dynamic simulations of the S1 protein-ACE2 receptor complex revealed the steric hindrance and Coulombic repulsion effects of different types ofN-glycans on the S1 protein interaction with ACE2. The results should contribute to future pathological studies of SARS-CoV-2 and therapeutic development of Covid-19, particularly using recombinant S1 proteins as models.Graphical abstractDisplay OmittedHighlights•SARS-CoV-2 interaction with ACE2 receptor is affected by glycosylation of S protein•De-glycosylated S1 receptor binding subunit showed stronger binding activity to ACE2•N-glycosylation profiles are different for S1 produced in different expression systems•N-glycans have steric hindrance and Coulombic repulsion effects to S protein bindingProtein; Virology; Structural biology; Computer simulation
