1. Structural Biology and Molecular Biophysics

Differential impact of BTK active site inhibitors on the conformational state of full-length BTK

  1. Raji E Joseph
  2. Neha Amatya
  3. D Bruce Fulton
  4. John R Engen
  5. Thomas E Wales  Is a corresponding author
  6. Amy Andreotti  Is a corresponding author
  1. Iowa State University, United States
  2. Northeastern University, United States
https://doi.org/10.7554/eLife.60470
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  1. Raji E Joseph
  2. Neha Amatya
  3. D Bruce Fulton
  4. John R Engen
  5. Thomas E Wales
  6. Amy Andreotti
(2020)
Differential impact of BTK active site inhibitors on the conformational state of full-length BTK
eLife 9:e60470.
https://doi.org/10.7554/eLife.60470
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Abstract

Bruton's tyrosine kinase (BTK) is targeted in the treatment of B-cell disorders including leukemias and lymphomas. Currently approved BTK inhibitors, including Ibrutinib, a first-in-class covalent inhibitor of BTK, bind directly to the kinase active site. While effective at blocking the catalytic activity of BTK, consequences of drug binding on the global conformation of full-length BTK are unknown. Here we uncover a range of conformational effects in full-length BTK induced by a panel of active site inhibitors, including large-scale shifts in the conformational equilibria of the regulatory domains. Additionally, we find that a remote Ibrutinib resistance mutation, T316A in the BTK SH2 domain, drives spurious BTK activity by destabilizing the compact autoinhibitory conformation of full-length BTK, shifting the conformational ensemble away from the autoinhibited form. Future development of BTK inhibitors will need to consider long-range allosteric consequences of inhibitor binding, including the emerging application of these BTK inhibitors in treating COVID-19.

Data availability

Hydrogen/deuterium exchange data have been deposited in the PRIDE database.

The following data sets were generated

Article and author information

Author details

  1. Raji E Joseph

    Biochemistry, Biophysics & Molecular Biology, Iowa State University, Ames, United States
    Competing interests
    The authors declare that no competing interests exist.

  2. Neha Amatya

    Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. D Bruce Fulton

    Biochemistry, Biophysics & Molecular Biology, Iowa State University, Ames, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. John R Engen

    Department of Chemistry and Chemical Biology, Northeastern University, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6918-9476

  5. Thomas E Wales

    Department of Chemistry and Chemical Biology, Northeastern University, Boston, United States
    For correspondence
    T.Wales@northeastern.edu
    Competing interests
    The authors declare that no competing interests exist.

  6. Amy Andreotti

    Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, United States
    For correspondence
    amyand@iastate.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6952-7244

Funding

National Institute of Allergy and Infectious Diseases (AI43957)

  • Amy Andreotti

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2020, Joseph et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Raji E Joseph
  2. Neha Amatya
  3. D Bruce Fulton
  4. John R Engen
  5. Thomas E Wales
  6. Amy Andreotti
(2020)
Differential impact of BTK active site inhibitors on the conformational state of full-length BTK
eLife 9:e60470.
https://doi.org/10.7554/eLife.60470

Share this article

https://doi.org/10.7554/eLife.60470

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Further reading

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics

    Impact of the clinically approved BTK inhibitors on the conformation of full-length BTK and analysis of the development of BTK resistance mutations in chronic lymphocytic leukemia

    Raji E Joseph, Thomas E Wales ... Amy H Andreotti
    Research Advance

    Inhibition of Bruton’s tyrosine kinase (BTK) has proven to be highly effective in the treatment of B-cell malignancies such as chronic lymphocytic leukemia (CLL), autoimmune disorders, and multiple sclerosis. Since the approval of the first BTK inhibitor (BTKi), Ibrutinib, several other inhibitors including Acalabrutinib, Zanubrutinib, Tirabrutinib, and Pirtobrutinib have been clinically approved. All are covalent active site inhibitors, with the exception of the reversible active site inhibitor Pirtobrutinib. The large number of available inhibitors for the BTK target creates challenges in choosing the most appropriate BTKi for treatment. Side-by-side comparisons in CLL have shown that different inhibitors may differ in their treatment efficacy. Moreover, the nature of the resistance mutations that arise in patients appears to depend on the specific BTKi administered. We have previously shown that Ibrutinib binding to the kinase active site causes unanticipated long-range effects on the global conformation of BTK (Joseph et al., 2020). Here, we show that binding of each of the five approved BTKi to the kinase active site brings about distinct allosteric changes that alter the conformational equilibrium of full-length BTK. Additionally, we provide an explanation for the resistance mutation bias observed in CLL patients treated with different BTKi and characterize the mechanism of action of two common resistance mutations: BTK T474I and L528W.

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    Small-Molecule Inhibitors: Disrupting enzyme fluidity

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