SARS-CoV-2 envelope protein topology in eukaryotic membranes
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Abstract
Coronavirus E protein is a small membrane protein found in the virus envelope. Different coronavirus E proteins share striking biochemical and functional similarities, but sequence conservation is limited. In this report, we studied the E protein topology from the new SARS-CoV-2 virus both in microsomal membranes and in mammalian cells. Experimental data reveal that E protein is a single-spanning membrane protein with the N-terminus being translocated across the membrane, while the C-terminus is exposed to the cytoplasmic side (Nt lum /Ct cyt ). The defined membrane protein topology of SARS-CoV-2 E protein may provide a useful framework to understand its interaction with other viral and host components and contribute to establish the basis to tackle the pathogenesis of SARS-CoV-2.
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SciScore for 10.1101/2020.05.27.118752: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Protein glycosylation status was analysed by Western Blot using an anti-c-myc antibody (Sigma), anti-rabbit IgG-peroxidase conjugated (Sigma), and with ECL developing reagent (GE Healthcare). anti-c-mycsuggested: Noneanti-rabbit IgG-peroxidasesuggested: NoneSoftware and Algorithms Sentences Resources All oligonucleotides were purchased from Macrogen (Seoul, South Korea). Macrogensuggested: (Macrogen, RRID:SCR_014454)Computer-assisted analysis of E protein sequence: Prediction of transmembrane … SciScore for 10.1101/2020.05.27.118752: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Protein glycosylation status was analysed by Western Blot using an anti-c-myc antibody (Sigma), anti-rabbit IgG-peroxidase conjugated (Sigma), and with ECL developing reagent (GE Healthcare). anti-c-mycsuggested: Noneanti-rabbit IgG-peroxidasesuggested: NoneSoftware and Algorithms Sentences Resources All oligonucleotides were purchased from Macrogen (Seoul, South Korea). Macrogensuggested: (Macrogen, RRID:SCR_014454)Computer-assisted analysis of E protein sequence: Prediction of transmembrane segments was done using up to 7 of the most common methods available on the Internet: ΔG Predictor [27,28] (http://dgpred.cbr.su.se/), TMHMM [29] (http://www.cbs.dtu.dk/services/TMHMM/), MEMSAT-SVM [30] (http://bioinf.cs.ucl.ac.uk/psipred/), TMpred (https://embnet.vital-it.ch/software/TMPRED_form.html), HMMTop [31] (http://www.enzim.hu/hmmtop/), Phobius [32] (http://phobius.sbc.su.se/) and TOPCONS [33] (http://topcons.net/). http://phobius.sbc.su.se/suggested: (Phobius, RRID:SCR_015643)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.
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