Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface

Wojcik, John; Lamontanara, Allan Joaquim; Grabe, Grzegorz; Koide, Akiko; Akin, Louesa; Gerig, Barbara; Hantschel, Oliver; Koide, Shohei

doi:10.1074/jbc.M115.707901

Title: Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface

Journal Article · Wed Feb 24 00:00:00 EST 2016 · Journal of Biological Chemistry

DOI:https://doi.org/10.1074/jbc.M115.707901· OSTI ID:1249239

Wojcik, John ^[1]; Lamontanara, Allan Joaquim ^[2]; Grabe, Grzegorz ^[3]; Koide, Akiko ^[1]; Akin, Louesa ^[1]; Gerig, Barbara ^[2]; Hantschel, Oliver ^[2]; Koide, Shohei ^[4]

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)

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.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Inst. of Health

Grant/Contract Number:: AC02-06CH11357; GM090324; P41 GM103403; T32GM07281; P30CA014599; KLS-3132-02-2013; KLS-3595-02-2015; S10 RR029205

OSTI ID:: 1249239

Journal Information:: Journal of Biological Chemistry, Vol. 291, Issue 16; ISSN 0021-9258

Publisher:: American Society for Biochemistry and Molecular BiologyCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 24 works

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Btk SH2-kinase interface is critical for allosteric kinase activation and its targeting inhibits B-cell neoplasms Duarte, Daniel P.; Lamontanara, Allan J.; La Sala, Giuseppina Nature Communications, Vol. 11, Issue 1 https://doi.org/10.1038/s41467-020-16128-5	journal	May 2020
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59 BASIC BIOLOGICAL SCIENCES
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
ABL tyrosine kinase
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protein engineering
protein-protein interaction
Src homology 2 domain (SH2 domain)
x-ray crystallography
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PPI inhibitor

Title: Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface

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