The Glycan-Specificity of the Pineapple Lectin AcmJRL and its Carbohydrate-Dependent Binding of the SARS-CoV-2 Spike Protein

  1. Joscha Meiers
  2. Jan Dastbaz
  3. Sebastian Adam
  4. Sari Rasheed
  5. Susanne H. Kirsch
  6. Peter Meiser
  7. Peter Gross
  8. Rolf Müller
  9. Alexander Titz

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Abstract

The current SARS-CoV-2 pandemic has become one of the most challenging global health threats, with over 530 million reported infections by May 2022. In addition to vaccines, research and development have also been directed towards novel drugs. Since the highly glycosylated spike protein of SARS-CoV-2 is essential for infection, it constitutes a prime target for antiviral agents. The pineapple-derived jacalin-related lectin (AcmJRL) is present in the medication bromelain in significant quantities and has previously been described to bind mannosides. Here, we elucidated its ligand specificity by glycan array analysis, quantified the interaction with carbohydrates and validated high-mannose glycans as preferred ligands. Because the SARS-CoV-2 spike protein was previously reported to carry a high proportion of high-mannose N-glycans, we tested the binding of AcmJRL to recombinantly produced spike protein. We could demonstrate that AcmJRL binds the spike protein with a low micromolar K D in a carbohydrate-dependent fashion, suggesting its use as a potential SARS-CoV-2 neutralising agent.

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    SciScore for 10.1101/2022.05.27.493400: (What is this?)

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

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    The mammalian cell line HEK 293/T served as host for recombinant production of the glycoprotein.
    HEK 293/T
    suggested: None
    Recombinant DNA
    SentencesResources
    , pCAGGS based, NCBI accession number: LT727518) was chosen for the mammalian expression system.
    pCAGGS
    suggested: RRID:Addgene_127347)
    The amplicon was digested with the respective restriction enzymes (ThermoFisher) and ligated with T4 Ligase (ThermoFisher) into the linearised πα-SHP-H vector to clone πα-SHP-H–Sgene with an N-terminal octahistidin tag.
    πα-SHP-H
    suggested: None
    Software and Algorithms
    SentencesResources
    Raw data (dot mean fluorescence intensity) was processed by GraphPad Prism 9 (GraphPad Software, USA)
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)

    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.

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

    About SciScore

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  2. Version published to 10.1101/2022.05.27.493400v1 on bioRxiv