Abstract
Several key mutations in the Spike protein receptor binding domain (RBD) have been identified to influence its affinity for the human Angiotensin-Converting Enzyme 2 (ACE2). Here, we perform a comparative study of the ACE2 binding to the wild type (Wuhan) RBD and some of its variants: Alpha B.1.1.7, Beta B.1.351, Delta B.1.617.2, Kappa B.1.617.1, B.1.1.7 + L452R and Omicron B.1.1.529. Using a coiled-coil mediated tethering approach of ACE2 in a novel surface plasmon resonance (SPR)-based assay, we measured interactions at different temperatures. Binding experiments at 10 °C enhanced the kinetic dissimilarities between the RBD variants and allowed a proper fit to a Langmuir 1:1 model with high accuracy and reproducibility, thus unraveling subtle differences within RBD mutants and ACE2 glycovariants. Our study emphasizes the importance of SPR-based assay parameters in the acquisition of biologically relevant data and offers a powerful tool to deepen our understanding of the role of the various RBD mutations in ACE2 interaction binding parameters.
【초록키워드】 ACE2, Mutation, B.1.351, variant, Delta, B.1.617.2, surface plasmon resonance, omicron, Spike protein, Receptor binding domain, Protein, L452R, B.1.1.7, RBD, B.1.617.1, Wuhan, Alpha, Beta, B.1.1.529, mutant, experiment, wild type, parameters, Spike protein receptor binding domain, binding, Human angiotensin-converting enzyme 2, Interaction, angiotensin, ACE2 binding, kinetic, surface plasmon, human Angiotensin-converting enzyme, powerful tool, reproducibility, high accuracy, RBD variants, acquisition, offer, parameter, approach, RBD mutation, dissimilarity, the RBD, the Spike, 1:1, 【제목키워드】 variant, Receptor binding domain, human ACE2, Impact, temperature, Interaction, the SARS-CoV-2,