Crystal structure of inhibitor-bound human MSPL that can activate high pathogenic avian influenza

  1. Ayako Ohno
  2. Nobuo Maita
  3. Takanori Tabata
  4. Hikaru Nagano
  5. Kyohei Arita
  6. Mariko Ariyoshi
  7. Takayuki Uchida
  8. Reiko Nakao
  9. Anayt Ulla
  10. Kosuke Sugiura
  11. Koji Kishimoto
  12. Shigetada Teshima-Kondo
  13. Yuushi Okumura
  14. Takeshi Nikawa

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Abstract

Infection of certain influenza viruses is triggered when its HA is cleaved by host cell proteases such as proprotein convertases and type II transmembrane serine proteases (TTSP). HA with a monobasic motif is cleaved by trypsin-like proteases, including TMPRSS2 and HAT, whereas the multibasic motif found in high pathogenicity avian influenza HA is cleaved by furin, PC5/6, or MSPL. MSPL belongs to the TMPRSS family and preferentially cleaves [R/K]-K-K-R↓ sequences. Here, we solved the crystal structure of the extracellular region of human MSPL in complex with an irreversible substrate-analog inhibitor. The structure revealed three domains clustered around the C-terminal α-helix of the SPD. The inhibitor structure and its putative model show that the P1-Arg inserts into the S1 pocket, whereas the P2-Lys and P4-Arg interacts with the Asp/Glu-rich 99-loop that is unique to MSPL. Based on the structure of MSPL, we also constructed a homology model of TMPRSS2, which is essential for the activation of the SARS-CoV-2 spike protein and infection. The model may provide the structural insight for the drug development for COVID-19.

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  1. Version published to 10.26508/lsa.202000849
  2. Version published to 10.1101/2020.06.12.149229v3 on bioRxiv
  3. Version published to 10.1101/2020.06.12.149229v2 on bioRxiv
  4. ScreenIT

    SciScore for 10.1101/2020.06.12.149229: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Software and Algorithms
    SentencesResources
    Complex formation, crystallization, and data collection: The peptide inhibitor (decanoyl-RVKR-cmk) was purchased from Merck-Millipore and reconstituted in dimethyl sulfoxide (DMSO).
    Merck-Millipore
    suggested: None
    Diffraction data were processed using the program iMosflm (31), followed by Aimless (32).
    iMosflm
    suggested: (iMosflm , RRID:SCR_014217)
    The model of SPD was manually fixed with COOT (34) and refined with Refmac5 (35).
    COOT
    suggested: (Coot, RRID:SCR_014222)
    All the structures in the figures were prepared using PyMOL (http://www.pymol.org/).
    PyMOL
    suggested: (PyMOL, RRID:SCR_000305)
    Homology modelling of TMPRSS2: The sequence alignment of the extracellular region of MSPL and TMPRSS2 was obtained using the BLAST webserver (https://www.uniprot.org/blast/).
    BLAST
    suggested: (BLASTX, RRID:SCR_001653)
    The homology model of TMPRSS2 was build using MODELLER (37).
    MODELLER
    suggested: (MODELLER, RRID:SCR_008395)

    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: 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: Please consider improving the rainbow (“jet”) colormap(s) used on pages 25 and 22. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    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.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  5. Version published to 10.1101/2020.06.12.149229v1 on bioRxiv