Impact of temperature on the affinity of SARS-CoV-2 Spike glycoprotein for host ACE2

Prévost, Jérémie; Richard, Jonathan; Gasser, Romain; Ding, Shilei; Fage, Clément; Anand, Sai Priya; Adam, Damien; Vergara, Natasha Gupta; Tauzin, Alexandra; Benlarbi, Mehdi; Gong, Shang Yu; Goyette, Guillaume; Privé, Anik; Moreira, Sandrine; Charest, Hugues; Roger, Michel; Mothes, Walther; Pazgier, M.; Brochiero, Emmanuelle; Boivin, Guy; Abrams, Cameron F.; Schön, Arne; Finzi, Andrés

doi:10.1016/j.jbc.2021.101151

Cited by 52 publications

(63 citation statements)

References 87 publications

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“…Healthy donors' plasmas, collected before the pandemic, were used as negative controls in flow cytometry and neutralization assays. The conformationally independent S2-specific monoclonal antibody CV3-25 [22,23] was used as a positive control and to normalize Spike expression in our flow cytometry assays, as described [22,24,25]. ACE2 binding was measured using the recombinant ACE2-Fc protein, which is composed of two ACE2 ectodomains linked to the Fc portion of the human IgG [26].…”

Section: Plasmas and Antibodiesmentioning

confidence: 99%

“…CV3-25 was used to normalize Spike expression, as reported [24]. The Median Fluorescence intensities (MFI) obtained with ACE2-Fc or plasma Abs were normalized to the MFI obtained with the conformationally and temperature independent CV3-25 anti-S2 antibody [22,23,29] and presented as ratio of the CV3-25-normalized values obtained with the D614G Spike.…”

Section: Cell Surface Staining and Flow Cytometry Analysismentioning

confidence: 99%

“…Plasmids expressing the different SARS-CoV-2 full Spikes were transfected into HEK 293T cells. Spike expression was normalized with the conformationally independent, S2-specific CV3-25 monoclonal antibody, as described [22][23][24]29]. All Spikes tested bound significantly better to ACE2 compared to their D614G counterpart, with Beta presenting the higher binding (Figure 1A and Table S2).…”

Section: Impact Of Temperature On Ace2 Binding By Sars-cov-2 Spikementioning

confidence: 99%

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Antigenicity of the Mu (B.1.621) and A.2.5 SARS-CoV-2 Spikes

Chatterjee

Tauzin

Laumaea

et al. 2022

Viruses

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The rapid emergence of SARS-CoV-2 variants is fueling the recent waves of the COVID-19 pandemic. Here, we assessed ACE2 binding and antigenicity of Mu (B.1.621) and A.2.5 Spikes. Both these variants carry some mutations shared by other emerging variants. Some of the pivotal mutations such as N501Y and E484K in the receptor-binding domain (RBD) detected in B.1.1.7 (Alpha), B.1.351 (Beta) and P.1 (Gamma) are now present within the Mu variant. Similarly, the L452R mutation of B.1.617.2 (Delta) variant is present in A.2.5. In this study, we observed that these Spike variants bound better to the ACE2 receptor in a temperature-dependent manner. Pseudoviral particles bearing the Spike of Mu were similarly neutralized by plasma from vaccinated individuals than those carrying the Beta (B.1.351) and Delta (B.1.617.2) Spikes. Altogether, our results indicate the importance of measuring critical parameters such as ACE2 interaction, plasma recognition and neutralization ability of each emerging variant.

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Section: Plasmas and Antibodiesmentioning

confidence: 99%

Section: Cell Surface Staining and Flow Cytometry Analysismentioning

confidence: 99%

Section: Impact Of Temperature On Ace2 Binding By Sars-cov-2 Spikementioning

confidence: 99%

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Antigenicity of the Mu (B.1.621) and A.2.5 SARS-CoV-2 Spikes

Chatterjee

Tauzin

Laumaea

et al. 2022

Viruses

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“…We observed that plasma recognition at the surface of infected (N+) cells significantly correlated with the binding of 293T cells expressing the full-length SARS-CoV-2 Spike (Figure 3A,D,G). Similarly, we observed significant correlations with neutralization potency using a well-established pseudoviral assay (Figure 3B,E,H) [4,5,11,18,23,24,33]. Since emerging evidence points to the potential benefits of antibody-mediated effector functions [18][19][20], we also evaluated whether ADCC correlated with recognition of the Spike at the surface of the infected cells.…”

Section: Recognition Of Sars-cov-2 Infected Paecs Correlates With Spike Recognition At the Surface Of 293t Cells Pseudoviral Neutralizatimentioning

confidence: 57%

“…Pre-coupled anti-nucleocapsid (anti-N) antibodies were used to distinguish infected from uninfected bystander cells (Figure 1). Using the conformational independent SARS-CoV-2 S2-specific CV3-25 antibody [18,[23][24][25][26], we detected high levels of the Spike at the surface of the infected N+ cells. CV3-25 specifically bound to infected (N+) pAECs when the cells were infected with the authentic D614G (Figure 1B) or Alpha (B.1.1.7) variant of concern (Figure 1C).…”

Section: Spike Recognition At the Surface Of Infected Primary Human Airway Epithelial Cellsmentioning

confidence: 96%

SARS-CoV-2 Spike Expression at the Surface of Infected Primary Human Airway Epithelial Cells

Ding

Adam

Beaudoin-Bussières

et al. 2021

Viruses

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Different serological assays were rapidly generated to study humoral responses against the SARS-CoV-2 Spike glycoprotein. Due to the intrinsic difficulty of working with SARS-CoV-2 authentic virus, most serological assays use recombinant forms of the Spike glycoprotein or its receptor binding domain (RBD). Cell-based assays expressing different forms of the Spike, as well as pseudoviral assays, are also widely used. To evaluate whether these assays recapitulate findings generated when the Spike is expressed in its physiological context (at the surface of the infected primary cells), we developed an intracellular staining against the SARS-CoV-2 nucleocapsid (N) to distinguish infected from uninfected cells. Human airway epithelial cells (pAECs) were infected with authentic SARS-CoV-2 D614G or Alpha variants. We observed robust cell-surface expression of the SARS-CoV-2 Spike at the surface of the infected pAECs using the conformational-independent anti-S2 CV3-25 antibody. The infected cells were also readily recognized by plasma from convalescent and vaccinated individuals and correlated with several serological assays. This suggests that the antigenicity of the Spike present at the surface of the infected primary cells is maintained in serological assays involving expression of the native full-length Spike.

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Evolutionary trajectory of receptor binding specificity and promiscuity of the spike protein of SARS‐CoV‐2

et al. 2022

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SARS‐CoV‐2 infects cells by attachment to its receptor—the angiotensin converting enzyme 2 (ACE2). Regardless of the wealth of structural data, little is known about the physicochemical mechanism of interactions of the viral spike (S) protein with ACE2 and how this mechanism has evolved during the pandemic. Here, we applied experimental and computational approaches to characterize the molecular interaction of S proteins from SARS‐CoV‐2 variants of concern (VOC). Data on kinetics, activation‐, and equilibrium thermodynamics of binding of the receptor binding domain (RBD) from VOC with ACE2 as well as data from computational protein electrostatics revealed a profound remodeling of the physicochemical characteristics of the interaction during the evolution. Thus, as compared to RBDs from Wuhan strain and other VOC, Omicron RBD presented as a unique protein in terms of conformational dynamics and types of non‐covalent forces driving the complex formation with ACE2. Viral evolution resulted in a restriction of the RBD structural dynamics, and a shift to a major role of polar forces for ACE2 binding. Further, we investigated how the reshaping of the physicochemical characteristics of interaction affects the binding specificity of S proteins. Data from various binding assays revealed that SARS‐CoV‐2 Wuhan and Omicron RBDs manifest capacity for promiscuous recognition of unrelated human proteins, but they harbor distinct reactivity patterns. These findings might contribute for mechanistic understanding of the viral tropism and capacity to evade immune responses during evolution.

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Impact of temperature on the affinity of SARS-CoV-2 Spike glycoprotein for host ACE2

Cited by 52 publications

References 87 publications

Antigenicity of the Mu (B.1.621) and A.2.5 SARS-CoV-2 Spikes

Antigenicity of the Mu (B.1.621) and A.2.5 SARS-CoV-2 Spikes

SARS-CoV-2 Spike Expression at the Surface of Infected Primary Human Airway Epithelial Cells

Evolutionary trajectory of receptor binding specificity and promiscuity of the spike protein of SARS‐CoV‐2

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