scholarly journals Biochemical, Biophysical, and Functional Characterization of Bacterially Expressed and Refolded Receptor Binding Domain ofPlasmodium vivaxDuffy-binding Protein

2001 ◽  
Vol 276 (20) ◽  
pp. 17111-17116 ◽  
Author(s):  
Sanjay Singh ◽  
Kailash Pandey ◽  
Rana Chattopadhayay ◽  
Syed Shams Yazdani ◽  
Andrew Lynn ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Binding Protein ◽  
Functional Characterization ◽  
Binding Domain ◽  
Receptor Binding Domain

Receptor-binding domain of human α2-macroglobulin Expression, folding and biochemical characterization of a high-affinity recombinant derivative

FEBS Letters ◽  
1994 ◽  
Vol 344 (2-3) ◽  
pp. 242-246 ◽  
Author(s):  
Thor Las Holtet ◽  
Kåre Lehmann Nielsen ◽  
Michael Etzerodt ◽  
Søren Kragh Moestrup ◽  
Jørgen Gliemann ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Biochemical Characterization ◽  
Binding Domain ◽  
Receptor Binding Domain ◽  
High Affinity ◽  
Α2 Macroglobulin

scholarly journals Structures and function of locked conformations of SARS-CoV-2 spike

2021 ◽  
Author(s):  
Kun Qu ◽  
Xiaoli Xiong ◽  
Katarzyna A. Ciazynska ◽  
Andrew P. Carter ◽  
John A. G. Briggs
Keyword(s):  
Receptor Binding ◽  
Functional Characterization ◽  
Protein S ◽  
Binding Domain ◽  
Receptor Binding Domain ◽  
Neutral Ph ◽  
And Function ◽  
Cellular Compartments ◽  
Domain Dynamics ◽  
Spike Dynamics

AbstractThe spike protein (S) of SARS-CoV-2 has been observed in three distinct pre-fusion conformations: locked, closed and open. Of these, the locked conformation was not previously observed for SARS-CoV-1 S and its function remains poorly understood. Here we engineered a SARS-CoV-2 S protein construct “S-R/x3” to arrest SARS-CoV-2 spikes in the locked conformation by a disulfide bond. Using this construct we determined high-resolution structures revealing two distinct locked states, with or without the D614G substitution that has become fixed in the globally circulating SARS-CoV-2 strains. The D614G mutation induces a structural change in domain D from locked-1 to locked-2 conformation to alter spike dynamics, promoting transition into the closed conformation from which opening of the receptor binding domain is permitted. The transition from locked to closed conformations is additionally promoted by a change from low to neutral pH. We propose that the locked conformations of S are present in the acidic cellular compartments where virus is assembled and egresses. In this model, release of the virion into the neutral pH extracellular space would favour transition to the closed form which itself can stochastically transition into the open form. The S-R/x3 construct provides a tool for the further structural and functional characterization of the locked conformations of S, as well as how sequence changes might alter S assembly and regulation of receptor binding domain dynamics.

scholarly journals An antibody-escape calculator for mutations to the SARS-CoV-2 receptor-binding domain

2021 ◽  
Author(s):  
Allison J Greaney ◽  
Tyler N Starr ◽  
Jesse D Bloom
Keyword(s):  
Receptor Binding ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Binding Domain ◽  
Receptor Binding Domain ◽  
Experimental Characterization ◽  
Antibody Recognition ◽  
Antigenic Properties ◽  
Interactive Version

A key goal of SARS-CoV-2 surveillance is to rapidly identify viral variants with mutations that reduce neutralization by polyclonal antibodies elicited by vaccination or infection. Unfortunately, direct experimental characterization of new viral variants lags their sequence-based identification. Here we help address this challenge by aggregating deep mutational scanning data into an "escape calculator" that estimates the antigenic effects of arbitrary combinations of mutations to the virus's spike receptor-binding domain (RBD). The calculator can be used to intuitively visualize how mutations impact polyclonal antibody recognition, and score the expected antigenic effect of combinations of mutations. These scores correlate with neutralization assays performed on SARS-CoV-2 variants, and emphasize the ominous antigenic properties of the recently described Omicron variant. An interactive version of the calculator is at https://jbloomlab.github.io/SARS2_RBD_Ab_escape_maps/escape-calc/, and we provide a Python module for batch processing.

scholarly journals The nuts and bolts of SARS-CoV-2 Spike Receptor Binding Domain heterologous expression

2021 ◽  
Author(s):  
Mariano Maffei ◽  
Linda C Montemiglio ◽  
Grazia Vitagliano ◽  
Luigi Fedele ◽  
Shaila Sellathurai ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Neutralizing Antibodies ◽  
Gel Filtration ◽  
Binding Domain ◽  
Model Systems ◽  
Receptor Binding Domain ◽  
Binding Ability ◽  
Hek 293 ◽  
293 Cells

COVID-19 is a highly infectious disease caused by a newly emerged coronavirus (SARS-CoV-2) that has rapidly progressed into a pandemic. This unprecedent emergency has stressed the significance of developing effective therapeutics to fight current and future outbreaks. The receptor-binding domain (RBD) of the SARS-CoV-2 surface Spike protein is the main target for vaccines and represents a helpful tool to produce neutralizing antibodies or diagnostic kits. In this work, we provide a detailed characterization of the native RBD produced in three major model systems: Escherichia coli, insect and HEK-293 cells. Circular dichroism, gel filtration chromatography and thermal denaturation experiments indicated that recombinant SARS-CoV-2 RBD proteins are stable and correctly folded. In addition, their functionality and receptor-binding ability were further evaluated through ELISA, flow cytometry assays and bio-layer interferometry.

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