Mutation N501Y in RBD of Spike Protein Strengthens the Interaction between COVID-19 and its Receptor ACE2
biorxiv(2021)
摘要
SARS-CoV-2 is spreading around the world for the past year. Enormous efforts have been taken to understand its mechanism of transmission. It is well established now that the receptor-binding domain (RBD) of the spike protein binds to the human angiotensin-converting enzyme 2 (ACE2) as its first step of entry. Being a single-stranded RNA virus, SARS-CoV-2 is evolving rapidly. Recently, several variants such as B.1.1.7, B.1.351, and P.1, with a key mutation N501Y on the RBD, appear to be more infectious to humans. To understand its mechanism, we combined cell surface binding assay, kinetics study, single-molecule technique, and computational method to investigate the interaction between these RBD (mutations) and ACE2. Remarkably, RBD with the N501Y mutation exhibited a considerably stronger interaction characterized from all these methodologies, while the other two mutations from B.1.351 contributed to a less effect. Fluorescence-activated cell scan (FACS) assays found that RBD N501Y mutations are of higher binding affinity to ACE2 than the wild type. Surface plasmon resonance further indicated that N501Y mutation had a faster association rate and slower dissociation rate. Consistent with the kinetics study, atomic force microscopy-based single-molecule force microscopy quantify their strength on living cells, showing a higher binding probability and unbinding force for the mutation. Finally, Steered Molecular Dynamics (SMD) simulations on the dissociation of RBD-ACE2 complexes revealed that the N501Y introduced additional π-π and π-cation interaction for the higher force/interaction. Taken together, we suggested that the reinforced interaction from N501Y mutation in RBD should play an essential role in the higher transmission of COVID-19 variants.
### Competing Interest Statement
The authors have declared no competing interest.
* (COVID-19)
: Coronavirus disease-19;
(SARS-CoV-2)
: Severe acute respiratory syndrome coronavirus-2;
(RBD)
: Receptor-binding domain
(MERS-CoV)
: Middle East respiratory syndrome coronavirus;
(ACE2)
: Angiotensin-converting enzyme 2;
(SPR)
: Surface Plasmon Resonance;
(FACS)
: Fluorescence-activated cell Scan;
(AFM)
: Atomic Force Microscopy;
(SMFS)
: Single-molecule force spectroscopy;
(SMD)
: Steered Molecular Dynamics.
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关键词
spike protein,mutation n501y,rbd,receptor
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