Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface
Abstract
Bcr-Abl is a constitutively active kinase that causes chronic myelogenous leukemia. We have shown that a tandem fusion of two designed binding proteins, termed monobodies, directed to the interaction interface between the Src homology 2 (SH2) and kinase domains and to the phosphotyrosine-binding site of the SH2 domain, respectively, inhibits the Bcr-Abl kinase activity. Because the latter monobody inhibits processive phosphorylation by Bcr-Abl and the SH2-kinase interface is occluded in the active kinase, it remained undetermined whether targeting the SH2-kinase interface alone was sufficient for Bcr-Abl inhibition. To address this question, we generated new, higher affinity monobodies with single nanomolar KD values targeting the kinase-binding surface of SH2. Structural and mutagenesis studies revealed the molecular underpinnings of the monobody-SH2 interactions. Importantly, the new monobodies inhibited Bcr-Abl kinase activity in vitro and in cells, and they potently induced cell death in chronic myelogenous leukemia cell lines. Lastly, this work provides strong evidence for the SH2-kinase interface as a pharmacologically tractable site for allosteric inhibition of Bcr-Abl.
- Authors:
-
- Univ. of Chicago, IL (United States)
- Ecole Polytechnique Federale Lausanne (Switzerland)
- Univ. of Chicago, IL (United States); Univ. of Gdansk (Poland)
- Univ. of Chicago, IL (United States); New York Univ. Langone Medical Center, NY (United States)
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Inst. of Health
- OSTI Identifier:
- 1249239
- Grant/Contract Number:
- AC02-06CH11357; GM090324; P41 GM103403; T32GM07281; P30CA014599; KLS-3132-02-2013; KLS-3595-02-2015; S10 RR029205
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Biological Chemistry
- Additional Journal Information:
- Journal Volume: 291; Journal Issue: 16; Journal ID: ISSN 0021-9258
- Publisher:
- American Society for Biochemistry and Molecular Biology
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; ABL tyrosine kinase; enzyme inhibitor; protein engineering; protein-protein interaction; Src homology 2 domain (SH2 domain); x-ray crystallography; FN3; PPI inhibitor
Citation Formats
Wojcik, John, Lamontanara, Allan Joaquim, Grabe, Grzegorz, Koide, Akiko, Akin, Louesa, Gerig, Barbara, Hantschel, Oliver, and Koide, Shohei. Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface. United States: N. p., 2016.
Web. doi:10.1074/jbc.M115.707901.
Wojcik, John, Lamontanara, Allan Joaquim, Grabe, Grzegorz, Koide, Akiko, Akin, Louesa, Gerig, Barbara, Hantschel, Oliver, & Koide, Shohei. Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface. United States. https://doi.org/10.1074/jbc.M115.707901
Wojcik, John, Lamontanara, Allan Joaquim, Grabe, Grzegorz, Koide, Akiko, Akin, Louesa, Gerig, Barbara, Hantschel, Oliver, and Koide, Shohei. Wed .
"Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface". United States. https://doi.org/10.1074/jbc.M115.707901. https://www.osti.gov/servlets/purl/1249239.
@article{osti_1249239,
title = {Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface},
author = {Wojcik, John and Lamontanara, Allan Joaquim and Grabe, Grzegorz and Koide, Akiko and Akin, Louesa and Gerig, Barbara and Hantschel, Oliver and Koide, Shohei},
abstractNote = {Bcr-Abl is a constitutively active kinase that causes chronic myelogenous leukemia. We have shown that a tandem fusion of two designed binding proteins, termed monobodies, directed to the interaction interface between the Src homology 2 (SH2) and kinase domains and to the phosphotyrosine-binding site of the SH2 domain, respectively, inhibits the Bcr-Abl kinase activity. Because the latter monobody inhibits processive phosphorylation by Bcr-Abl and the SH2-kinase interface is occluded in the active kinase, it remained undetermined whether targeting the SH2-kinase interface alone was sufficient for Bcr-Abl inhibition. To address this question, we generated new, higher affinity monobodies with single nanomolar KD values targeting the kinase-binding surface of SH2. Structural and mutagenesis studies revealed the molecular underpinnings of the monobody-SH2 interactions. Importantly, the new monobodies inhibited Bcr-Abl kinase activity in vitro and in cells, and they potently induced cell death in chronic myelogenous leukemia cell lines. Lastly, this work provides strong evidence for the SH2-kinase interface as a pharmacologically tractable site for allosteric inhibition of Bcr-Abl.},
doi = {10.1074/jbc.M115.707901},
journal = {Journal of Biological Chemistry},
number = 16,
volume = 291,
place = {United States},
year = {Wed Feb 24 00:00:00 EST 2016},
month = {Wed Feb 24 00:00:00 EST 2016}
}
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