Rise and Fall of SARS-CoV-2 Lineage A.27 in Germany

Here, we report on the increasing frequency of the SARS-CoV-2 lineage A.27 in Germany during the first months of 2021. Genomic surveillance identified 710 A.27 genomes in Germany as of 2 May 2021, with a vast majority identified in laboratories from a single German state (Baden-Wuerttemberg, n = 572; 80.5%). Baden-Wuerttemberg is located near the border with France, from where most A.27 sequences were entered into public databases until May 2021. The first appearance of this lineage based on sequencing in a laboratory in Baden-Wuerttemberg can be dated to early January ’21. From then on, the relative abundance of A.27 increased until the end of February but has since declined—meanwhile, the abundance of B.1.1.7 increased in the region. The A.27 lineage shows a mutational pattern typical of VOIs/VOCs, including an accumulation of amino acid substitutions in the Spike glycoprotein. Among those, L18F, L452R and N501Y are located in the epitope regions of the N-terminal- (NTD) or receptor binding domain (RBD) and have been suggested to result in immune escape and higher transmissibility. In addition, A.27 does not show the D614G mutation typical for all VOIs/VOCs from the B lineage. Overall, A.27 should continue to be monitored nationally and internationally, even though the observed trend in Germany was initially displaced by B.1.1.7 (Alpha), while now B.1.617.2 (Delta) is on the rise.

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Ten emerging SARS-CoV-2 spike variants exhibit variable infectivity, animal tropism, and antibody neutralization

Communications Biology ◽

10.1038/s42003-021-02728-4 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Li Zhang ◽

Zhimin Cui ◽

Qianqian Li ◽

Bo Wang ◽

Yuanling Yu ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Amino Acid ◽

Receptor Binding ◽

Immune Escape ◽

Cathepsin L ◽

Single Amino Acid ◽

Receptor Binding Domain ◽

Therapeutic Antibodies ◽

Amino Acid Mutations ◽

Better Than

AbstractEmerging mutations in SARS-CoV-2 cause several waves of COVID-19 pandemic. Here we investigate the infectivity and antigenicity of ten emerging SARS-CoV-2 variants—B.1.1.298, B.1.1.7(Alpha), B.1.351(Beta), P.1(Gamma), P.2(Zeta), B.1.429(Epsilon), B.1.525(Eta), B.1.526-1(Iota), B.1.526-2(Iota), B.1.1.318—and seven corresponding single amino acid mutations in the receptor-binding domain using SARS-CoV-2 pseudovirus. The results indicate that the pseudovirus of most of the SARS-CoV-2 variants (except B.1.1.298) display slightly increased infectivity in human and monkey cell lines, especially B.1.351, B.1.525 and B.1.526 in Calu-3 cells. The K417N/T, N501Y, or E484K-carrying variants exhibit significantly increased abilities to infect mouse ACE2-overexpressing cells. The activities of furin, TMPRSS2, and cathepsin L are increased against most of the variants. RBD amino acid mutations comprising K417T/N, L452R, Y453F, S477N, E484K, and N501Y cause significant immune escape from 11 of 13 monoclonal antibodies. However, the resistance to neutralization by convalescent serum or vaccines elicited serum is mainly caused by the E484K mutation. The convalescent serum from B.1.1.7- and B.1.351-infected patients neutralized the variants themselves better than other SARS-CoV-2 variants. Our study provides insights regarding therapeutic antibodies and vaccines, and highlights the importance of E484K mutation.

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SARS-CoV-2 Variant of Concern Substitutions Alter Spike Glycoprotein Receptor Binding Domain Structure and Stability

10.1101/2021.05.11.443443 ◽

2021 ◽

Author(s):

Daniel L Moss ◽

Jay Rappaport

Keyword(s):

Receptor Binding ◽

Rapid Development ◽

Structure And Function ◽

Binding Domain ◽

Amino Acid Substitutions ◽

Receptor Binding Domain ◽

Spike Glycoprotein ◽

Structure And Stability ◽

The World ◽

And Function

The emergence of severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) and the subsequent COVID-19 pandemic has significantly impacted the world not just with disease and death but also economic turmoil. The rapid development and deployment of extremely effective vaccines against SARS-CoV-2 has made the end of the pandemic a reality within reach. However, as the virus spreads it has acquired mutations; and thus, variants of concern have emerged that are more infectious and reduce the efficacy of existing vaccines. While promising efforts are underway to combat these variants, the evolutionary pressures leading to these variants are poorly understood. To that end, here we have studied the effects of three amino-acid substitutions on the structure and function of the SARS-CoV-2 spike glycoprotein receptor-binding domain found in several variants of concern such as B.1.1.7, B.1.351 and P.1 that are now circulating. We found that these substitutions alter the RBD structure and stability, as well as its ability to bind to ACE2, which may have opposing and compensatory effects. These findings provide new insights into how these Variants of Concern (VOC) may have been selected to optimize infectivity while maintaining the structure and stability of the receptor binding domain.

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Murine Coronavirus Evolution In Vivo: Functional Compensation of a Detrimental Amino Acid Substitution in the Receptor Binding Domain of the Spike Glycoprotein

Journal of Virology ◽

10.1128/jvi.79.12.7629-7640.2005 ◽

2005 ◽

Vol 79 (12) ◽

pp. 7629-7640 ◽

Cited By ~ 18

Author(s):

Sonia Navas-Martin ◽

Susan T. Hingley ◽

Susan R. Weiss

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Receptor Binding ◽

Amino Acid Substitution ◽

Binding Domain ◽

Receptor Binding Domain ◽

Spike Glycoprotein ◽

Functional Compensation

ABSTRACT Murine coronavirus A59 strain causes mild to moderate hepatitis in mice. We have previously shown that mutants of A59, unable to induce hepatitis, may be selected by persistent infection of primary glial cells in vitro. These in vitro isolated mutants encoded two amino acids substitutions in the spike (S) gene: Q159L lies in the putative receptor binding domain of S, and H716D, within the cleavage signal of S. Here, we show that hepatotropic revertant variants may be selected from these in vitro isolated mutants (Q159L-H716D) by multiple passages in the mouse liver. One of these mutants, hr2, was chosen for more in-depth study based on a more hepatovirulent phenotype. The S gene of hr2 (Q159L- R654H -H716D- E1035D ) differed from the in vitro isolates (Q159L-H716D) in only 2 amino acids (R654H and E1035D). Using targeted RNA recombination, we have constructed isogenic recombinant MHV-A59 viruses differing only in these specific amino acids in S (Q159L-R654H-H716D-E1035D). We demonstrate that specific amino acid substitutions within the spike gene of the hr2 isolate determine the ability of the virus to cause lethal hepatitis and replicate to significantly higher titers in the liver compared to wild-type A59. Our results provide compelling evidence of the ability of coronaviruses to rapidly evolve in vivo to highly virulent phenotypes by functional compensation of a detrimental amino acid substitution in the receptor binding domain of the spike glycoprotein.

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A Novel Real-Time RT-PCR-Based Methodology for the Preliminary Typing of SARS-CoV-2 Variants, Employing Non-Extendable LNA Oligonucleotides and Three Signature Mutations at the Spike Protein Receptor-Binding Domain

Life ◽

10.3390/life11101015 ◽

2021 ◽

Vol 11 (10) ◽

pp. 1015

Author(s):

Serafeim C. Chaintoutis ◽

Taxiarchis Chassalevris ◽

Sofia Balaska ◽

Evangelia Mouchtaropoulou ◽

George Tsiolas ◽

...

Keyword(s):

Amino Acid ◽

Real Time ◽

Receptor Binding ◽

Binding Domain ◽

Locked Nucleic Acid ◽

Spike Protein ◽

Amino Acid Substitutions ◽

Receptor Binding Domain ◽

Rt Pcr ◽

Protein Receptor

Mutations resulting in amino-acid substitutions of the SARS-CoV-2 spike protein receptor-binding domain (RBD) have been associated with enhanced transmissibility and immune escape of the respective variants, namely Alpha, Beta, Gamma or Delta. Rapid identification of the aforementioned variants of concern and their discrimination of other variants is thus of importance for public health interventions. For this reason, a one-step real-time RT-PCR assay employing four locked nucleic acid (LNA) modified TaqMan probes was developed, to target signature mutations associated with amino-acid substitutions at positions 478, 484 and 501 present in the receptor-binding motif (RBM) of the spike protein RBD. This region contains most contacting residues of SARS-CoV-2 that bind to ACE2. A novel strategy employing the use of non-extendable LNA oligonucleotide blockers that can reduce non-specific hybridization of probes increased the number of different mutated sites examined in a multiplex PCR. The combinatory analysis of the different fluorescence signals obtained enabled the preliminary differentiation of SARS-CoV-2 variants of concern. The assay is sensitive with a LOD of 263 copies/reaction for the Delta variant, 170 copies/reaction for the Beta variant, amplification efficiencies > 91% and a linear range of >5 log10 copies/reaction against all targets. Validation of the assay using known SARS-CoV-2-positive and negative samples from humans and animals revealed its ability to correctly identify the targeted mutations and preliminary characterize the SARS-CoV-2 variants. The novel approach for mutation typing using LNA oligonucleotide blockers can be modified to target signature mutations at four different sites in the RBM and further expand the range of variants detected.

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Structural analysis of novel amino acid substitutions in SARS-CoV-2 spike protein receptor-binding domain

Hacettepe Journal of Biology and Chemistry ◽

10.15671/hjbc.776430 ◽

2021 ◽

Author(s):

Burcu BİTERGE SÜT

Keyword(s):

Amino Acid ◽

Structural Analysis ◽

Receptor Binding ◽

Binding Domain ◽

Spike Protein ◽

Amino Acid Substitutions ◽

Receptor Binding Domain ◽

Protein Receptor

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Altered N-/O-Glycosylation Sites On The Receptor-Binding Domain (RBD) in COVID-19 Are Related To Coronavirus Infection And Pathogenesis.

10.21203/rs.3.rs-407527/v1 ◽

2021 ◽

Author(s):

Shiwei Wang ◽

Fan Xiao ◽

Lingling He ◽

Junru Yang ◽

Hongshan Wei

Keyword(s):

Amino Acid ◽

Receptor Binding ◽

Amino Acid Sequences ◽

Receptor Binding Domain ◽

Spike Glycoprotein ◽

Multiple Alignments ◽

Glycosylation Sites ◽

Coronavirus Infection ◽

Clustal Omega ◽

Bat Coronavirus

Abstract Objective: To discuss the possible significances of N-Linked and O-Linked glycosylation on the RBD of COVID-19Methods: Amino acid sequences multiple alignments of RBD used Clustal Omega (v.1.2.4) using default parameters. Prediction of potential N-linked glycosylation sites by NetNGlyc 1.0 server. Prediction of potential O-linked glycosylation sites by NetOGlyc 4.0 Server.Result: COVID-19 Spike glycoprotein has 22 potential N-linked glycosylation sites and 3 O-linked glycosylation sites. 2 of 22 N-linked glycosylation sites distributed in RBD. None of the 3 O-linked glycosylation sites distributed in RBD, which is markedly different from SARS and other bat coronavirus using ACE2 as a receptor. Comparing with its close coronavirus, but which can’t use ACE2 as a receptor, the COVID-19 has little N- and O-linked glycosylation sites.Conclusion: we show the obvious differences in glycosylation sites in RBD between COVID-19 and other coronaviruses. We speculate that altered N-/O-glycosylation sites on RBD in COVID-19 are related to its infection and pathogenesis.

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Role of key point Mutations in Receptor Binding Domain of SARS-CoV-2 Spike Glycoprotein

Infectious Disorders - Drug Targets ◽

10.2174/1871526520666200804161650 ◽

2020 ◽

Vol 20 ◽

Author(s):

Bipin Singh

Keyword(s):

Receptor Binding ◽

Point Mutations ◽

Binding Domain ◽

Receptor Binding Domain ◽

Spike Glycoprotein ◽

Angiotensin Converting Enzyme 2 ◽

Recent Outbreak ◽

Novel Coronavirus ◽

Genomic Similarity

: The recent outbreak of novel coronavirus (SARS-CoV-2 or 2019-nCoV) and its worldwide spread is posing one of the major threats to human health and the world economy. It has been suggested that SARS-CoV-2 is similar to SARSCoV based on the comparison of the genome sequence. Despite the genomic similarity between SARS-CoV-2 and SARSCoV, the spike glycoprotein and receptor binding domain in SARS-CoV-2 shows the considerable difference compared to SARS-CoV, due to the presence of several point mutations. The analysis of receptor binding domain (RBD) from recently published 3D structures of spike glycoprotein of SARS-CoV-2 (Yan, R., et al. (2020); Wrapp, D., et al. (2020); Walls, A. C., et al. (2020)) highlights the contribution of a few key point mutations in RBD of spike glycoprotein and molecular basis of its efficient binding with human angiotensin-converting enzyme 2 (ACE2).

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ReaxFF-based molecular dynamics simulation of the interaction between plasma ROS and the receptor-binding domain of the spike protein in the capsid protein of SARS-CoV-2

Journal of Physics D Applied Physics ◽

10.1088/1361-6463/ac360e ◽

2021 ◽

Author(s):

Huichao Wang ◽

Tong Zhao ◽

Shuhui Yang ◽

Liang Zou ◽

Xiaolong Wang ◽

...

Keyword(s):

Molecular Dynamics ◽

Amino Acid ◽

Capsid Protein ◽

Receptor Binding ◽

Hydrogen Abstraction ◽

Binding Domain ◽

Spike Protein ◽

Receptor Binding Domain ◽

Amino Acid Residues ◽

Global Epidemic

Abstract Under the severe situation of the current global epidemic, researchers have been working hard to find a reliable way to suppress the infection of the virus and prevent the spread of the epidemic. Studies have shown that the recognition and binding of human angiotensin-converting enzyme 2 (ACE2) by the receptor-binding domain (BRD) of spike protein on the surface of SARS-CoV-2 is a crucial step for SARS-CoV-2 to invade human receptor cells, and blocking this process can inhibit the virus from invading human normal cells. Plasma treatment can disrupt the structure of the RBD and effectively block the binding process. However, the mechanism by which plasma blocks the recognition and binding between the two is not clear. In this study, reaction process between reactive oxygen species (ROS) in plasma and the molecular model of RBD was simulated using a reactive molecular dynamics method. The results showed that the destruction of RBD molecule by ROS was triggered by hydrogen abstraction reactions. O and OH abstracted H atoms from RBD, while the H atoms of H2O2 and HO2 were abstracted by RBD. The hydrogen abstraction resulted in the breakage of C-H, N-H, O-H and C=O bonds and the formation of C=C, C=N bonds. The addition reaction of OH increased the number of O-H bonds and caused the formation of C-O, N-O and O-H bonds. The dissociation of N-H bonds led to the destruction of the original structure of peptide bonds and amino acid residues, change the type of amino acid residues, and caused the conversion of N-C and N=C, C=O and C-O. The simulation partially elucidated the microscopic mechanism of the interaction between ROS in plasma and the capsid protein of SARS-CoV-2, providing theoretical support for the control of SARS-CoV-2 infection by plasma, a contribution to overcoming the global epidemic problem.

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Amino acid substitutions in human growth hormone affect coiled-coil content and receptor binding

10.1101/2021.12.16.473085 ◽

2021 ◽

Author(s):

Andrei Rajkovic ◽

Sandesh Kanchugal ◽

Eldar Abdurakhmanov ◽

Rebecca Howard ◽

Astrid Gräslund ◽

...

Keyword(s):

Growth Hormone ◽

Amino Acid ◽

Binding Site ◽

Human Growth Hormone ◽

Receptor Binding ◽

Coiled Coil ◽

Human Growth ◽

Zinc Binding ◽

Amino Acid Substitutions ◽

Great Relevance

The interaction between human Growth Hormone (hGH) and hGH Receptor (hGHR) has great relevance to human diseases such as acromegaly and cancer. HGH has been extensively engineered by other workers to improve binding and other properties. We used a computational screen to select substitutions at single hGH positions within the hGHR-binding site. We find that, while many successfully slow down dissociation of the hGH-hGHR complex once bound, they also slow down the association of hGH to hGHR. We are particularly interested in E174 which belongs to the hGH zinc-binding triad, and which spans coiled-coil helices and obeys the coiled-coil heptad pattern. Surprisingly, substituting E174 with A leads to substantial increase in an experimental measure of coiled-coil content. E174A is known to increase affinity of hGH against hGHR; here we show that this is simply because the off-rate is slowed down more than the on-rate, in line with what has been found for other affinity-improving mutations. For E174Y (and mutations at other sites) the slowdown in on-rate was greater, leading to decreased affinity. The results point to a link between coiled-coiling, zinc binding, and hGHR-binding affinity in hGH, and also suggest rules for choosing affinity-increasing substitutions.

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