Site-specific steric control of SARS-CoV-2 spike glycosylation
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
A central tenet in the design of vaccines is the display of native-like antigens in the elicitation of protective immunity. The abundance of N-linked glycans across the SARS-CoV-2 spike protein is a potential source of heterogeneity between the many different vaccine candidates under investigation. Here, we investigate the glycosylation of recombinant SARS-CoV-2 spike proteins from five different laboratories and compare them against infectious virus S protein. We find patterns which are conserved across all samples and this can be associated with site-specific stalling of glycan maturation which act as a highly sensitive reporter of protein structure. Molecular dynamics (MD) simulations of a fully glycosylated spike support s a model of steric restrictions that shape enzymatic processing of the glycans. These results suggest that recombinant spike-based SARS-CoV-2 immunogen glycosylation reproducibly recapitulates signatures of viral glycosylation.
Article activity feed
-
SciScore for 10.1101/2021.03.08.433764: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
NIH rigor criteria are not applicable to paper type.Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Spike ectodomain was expressed by transient transfection HEK293T (ATCC, CRL-3216) cells for 6 days at 30 °C. HEK293Tsuggested: NoneS protein was expressed in HEK 293F cells. HEK 293Fsuggested: RRID:CVCL_6642)Software and Algorithms Sentences Resources The S protein concentration was determined by the Nanodrop method using the proteins peptidic molecular weight and extinction coefficient as determined by the online ExPASy software (ProtParam). ExPASysuggested: NoneIntegrative modelling and molecular dynamics simulation: The model of S protein was built using Modeller 9.2175 with three … SciScore for 10.1101/2021.03.08.433764: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
NIH rigor criteria are not applicable to paper type.Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Spike ectodomain was expressed by transient transfection HEK293T (ATCC, CRL-3216) cells for 6 days at 30 °C. HEK293Tsuggested: NoneS protein was expressed in HEK 293F cells. HEK 293Fsuggested: RRID:CVCL_6642)Software and Algorithms Sentences Resources The S protein concentration was determined by the Nanodrop method using the proteins peptidic molecular weight and extinction coefficient as determined by the online ExPASy software (ProtParam). ExPASysuggested: NoneIntegrative modelling and molecular dynamics simulation: The model of S protein was built using Modeller 9.2175 with three structural templates: i) the cryo-EM structure of SARS-CoV-2 S ECD in the open state (PDB: 6VSB)29, ii) the NMR structure of SAR-CoV S HR2 domain (PDB: 2FXP) 61, and iii) the NMR structure of HIV-1 gp-41 TM domain (PDB: 5JYN)62. Modellersuggested: (MODELLER, RRID:SCR_008395)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
-