Potent, multi-target serine protease inhibition achieved by a simplified β-sheet motif

Xingchen Chen, Blake T. Riley, Simon J. De Veer, David E. Hoke, Jessica Van Haeften, Darren Leahy, Joakim E. Swedberg, Maria Brattsand, Perry J. Hartfield, Ashley M. Buckle, Jonathan M. Harris

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Engagement of an extended β-sheet is a common substrate/inhibitor interaction at the active site of serine proteases and is an important feature of Laskowski mechanism inhibitors that present a substrate-like loop to a target protease. This loop is cleaved but subsequently relegated forming a stable inhibitor/protease complex. Laskowski inhibitors are ubiquitous in nature and are used extensively in serine protease inhibitor design. However, most studies concentrate on introducing new sidechain interactions rather than the direct contributions of the substrate-like β-sheet to enzyme inhibition. Here we report the crystal structure of an simplified β-sheet inhibitory motif within the Sunflower Trypsin Inhibitor (SFTI) in complex with trypsin. We show that the intramolecular hydrogen bond network of this SFTI variant (SFTI-TCTR) engages the inhibitor sidechains that would normally interact with a target protease, giving mainchain interactions a more prominent role in complex formation. Despite having reduced sidechain interactions, this SFTI variant is remarkably potent and inhibits a diverse range of serine proteases. Crystal structural analysis and molecular modelling of SFTI-TCTR complexes again indicates an interface dominated by β–sheet interactions, highlighting the importance of this motif and the adaptability of SFTI as a scaffold for inhibitor design.

Original languageEnglish
Article numbere0210842
Number of pages15
JournalPLoS ONE
Volume14
Issue number1
DOIs
Publication statusPublished - 22 Jan 2019

Keywords

  • serine proteases
  • proteases
  • hydrogen bonding
  • crystal structure
  • arginine
  • molecular dynamics
  • crystals
  • enzyme inhibitors

Cite this

Chen, X., Riley, B. T., De Veer, S. J., Hoke, D. E., Van Haeften, J., Leahy, D., ... Harris, J. M. (2019). Potent, multi-target serine protease inhibition achieved by a simplified β-sheet motif. PLoS ONE, 14(1), [e0210842]. https://doi.org/10.1371/journal.pone.0210842
Chen, Xingchen ; Riley, Blake T. ; De Veer, Simon J. ; Hoke, David E. ; Van Haeften, Jessica ; Leahy, Darren ; Swedberg, Joakim E. ; Brattsand, Maria ; Hartfield, Perry J. ; Buckle, Ashley M. ; Harris, Jonathan M. / Potent, multi-target serine protease inhibition achieved by a simplified β-sheet motif. In: PLoS ONE. 2019 ; Vol. 14, No. 1.
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Chen, X, Riley, BT, De Veer, SJ, Hoke, DE, Van Haeften, J, Leahy, D, Swedberg, JE, Brattsand, M, Hartfield, PJ, Buckle, AM & Harris, JM 2019, 'Potent, multi-target serine protease inhibition achieved by a simplified β-sheet motif' PLoS ONE, vol. 14, no. 1, e0210842. https://doi.org/10.1371/journal.pone.0210842

Potent, multi-target serine protease inhibition achieved by a simplified β-sheet motif. / Chen, Xingchen; Riley, Blake T.; De Veer, Simon J.; Hoke, David E.; Van Haeften, Jessica; Leahy, Darren; Swedberg, Joakim E.; Brattsand, Maria; Hartfield, Perry J.; Buckle, Ashley M.; Harris, Jonathan M.

In: PLoS ONE, Vol. 14, No. 1, e0210842, 22.01.2019.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Chen, Xingchen

AU - Riley, Blake T.

AU - De Veer, Simon J.

AU - Hoke, David E.

AU - Van Haeften, Jessica

AU - Leahy, Darren

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AU - Harris, Jonathan M.

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AB - Engagement of an extended β-sheet is a common substrate/inhibitor interaction at the active site of serine proteases and is an important feature of Laskowski mechanism inhibitors that present a substrate-like loop to a target protease. This loop is cleaved but subsequently relegated forming a stable inhibitor/protease complex. Laskowski inhibitors are ubiquitous in nature and are used extensively in serine protease inhibitor design. However, most studies concentrate on introducing new sidechain interactions rather than the direct contributions of the substrate-like β-sheet to enzyme inhibition. Here we report the crystal structure of an simplified β-sheet inhibitory motif within the Sunflower Trypsin Inhibitor (SFTI) in complex with trypsin. We show that the intramolecular hydrogen bond network of this SFTI variant (SFTI-TCTR) engages the inhibitor sidechains that would normally interact with a target protease, giving mainchain interactions a more prominent role in complex formation. Despite having reduced sidechain interactions, this SFTI variant is remarkably potent and inhibits a diverse range of serine proteases. Crystal structural analysis and molecular modelling of SFTI-TCTR complexes again indicates an interface dominated by β–sheet interactions, highlighting the importance of this motif and the adaptability of SFTI as a scaffold for inhibitor design.

KW - serine proteases

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KW - hydrogen bonding

KW - crystal structure

KW - arginine

KW - molecular dynamics

KW - crystals

KW - enzyme inhibitors

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Chen X, Riley BT, De Veer SJ, Hoke DE, Van Haeften J, Leahy D et al. Potent, multi-target serine protease inhibition achieved by a simplified β-sheet motif. PLoS ONE. 2019 Jan 22;14(1). e0210842. https://doi.org/10.1371/journal.pone.0210842