Structure and activity of human TMPRSS2 protease implicated in SARS-CoV-2 activation

Version published to 10.1038/s41589-022-01059-7

Jun 8, 2022

SciScore for 10.1101/2021.06.23.449282: (What is this?)

Please note, not all rigor criteria are appropriate for all manuscripts.

Table 1: Rigor

Ethics	not detected.
Sex as a biological variable	not detected.
Randomization	not detected.
Blinding	not detected.
Power Analysis	not detected.
Cell Line Authentication	not detected.

Table 2: Resources

Experimental Models: Cell Lines
Sentences	Resources
Baculovirus mediated dasTMPRSS2 protein production in Sf9 insect cells: Sf9 cells were grown in I-Max Insect Medium (Wisent Biocenter; Cat# 301-045-LL) to a density of 4×106 cells/mL and infected with 20 mL/L of suspension culture of baculovirus infected insect cells prior to incubation on an orbital shaker (145 rpm, 26 °C).	Sf9 suggested: None
Recombinant DNA
Sentences	Resources
Construct design and cloning: A construct encoding residues 109-492 comprising soluble …

SciScore for 10.1101/2021.06.23.449282: (What is this?)

Please note, not all rigor criteria are appropriate for all manuscripts.

Table 1: Rigor

Ethics	not detected.
Sex as a biological variable	not detected.
Randomization	not detected.
Blinding	not detected.
Power Analysis	not detected.
Cell Line Authentication	not detected.

Table 2: Resources

Experimental Models: Cell Lines
Sentences	Resources
Baculovirus mediated dasTMPRSS2 protein production in Sf9 insect cells: Sf9 cells were grown in I-Max Insect Medium (Wisent Biocenter; Cat# 301-045-LL) to a density of 4×106 cells/mL and infected with 20 mL/L of suspension culture of baculovirus infected insect cells prior to incubation on an orbital shaker (145 rpm, 26 °C).	Sf9 suggested: None
Recombinant DNA
Sentences	Resources
Construct design and cloning: A construct encoding residues 109-492 comprising soluble TMPRSS2 ectodomain was amplified by two PCR fragments (Addgene plasmid# 53887) and subcloned into the pFHMSP-LIC C donor plasmid by LIC method.	pFHMSP-LIC C suggested: RRID:Addgene_26100)
Software and Algorithms
Sentences	Resources
To determine Michaelis Menten kinetic parameters, 50 μL 4x enzyme stock was added through automated addition to microplates containing 150 μL substrate (0.5-1000 μM) in triplicate and initial reaction velocities were plotted against substrate concentration and curve fit using GraphPad Prism.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
Then, 7 inhibitor concentrations spanning three orders of magnitude across the IC50 value were used and inhibitor reaction velocities were normalized to uninhibited enzyme and plotted as one-site dose response curves in GraphPad.	GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
Human TTSP FASTA sequences (isoform 1) were accessed from UniProt and TMPRSS2 orthologs identified with UniProt BLAST.	UniProt suggested: (UniProtKB, RRID:SCR_004426) BLAST suggested: (BLASTX, RRID:SCR_001653)
Protein Visualization and Property Calculation: The structure of dasTMPRSS2 was inspected and compared to other TTSPs using PyMol (Schrodinger) and the Molecular Operating Environment (	PyMol suggested: (PyMOL, RRID:SCR_000305)

Results from OddPub: Thank you for sharing your data.

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: We found the following clinical trial numbers in your paper:

Identifier	Status	Title
NCT04583592	Completed	Camostat Efficacy vs. Placebo for Outpatient Treatment of CO…
NCT04625114	Recruiting	The Potential of Oral Camostat in Early COVID-19 Disease in …
NCT04418128	Not yet recruiting	Clinical Efficacy of Nafamostat Mesylate for COVID-19 Pneumo…
NCT04473053	Recruiting	Rapid Experimental Medicine for COVID-19

Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

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.

Results from scite Reference Check: We found no unreliable references.

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Structure and activity of human TMPRSS2 protease implicated in SARS-CoV-2 activation

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