Format

Send to

Choose Destination
J Biol Chem. 2016 Apr 15;291(16):8836-47. doi: 10.1074/jbc.M115.707901. Epub 2016 Feb 24.

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

Author information

1
From the Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois 60637.
2
Swiss Institute for Experimental Cancer Research, School of Life Sciences, École polytechnique fédérale de Lausanne, 1015 Lausanne, Switzerland, and.
3
From the Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois 60637, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Gdansk, Poland.
4
Swiss Institute for Experimental Cancer Research, School of Life Sciences, École polytechnique fédérale de Lausanne, 1015 Lausanne, Switzerland, and oliver.hantschel@epfl.ch.
5
From the Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois 60637, Shohei.Koide@nyumc.org.

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. This work provides strong evidence for the SH2-kinase interface as a pharmacologically tractable site for allosteric inhibition of Bcr-Abl.

KEYWORDS:

ABL tyrosine kinase; FN3; PPI inhibitor; Src homology 2 domain (SH2 domain); enzyme inhibitor; protein engineering; protein-protein interaction; x-ray crystallography

PMID:
26912659
PMCID:
PMC4861451
DOI:
10.1074/jbc.M115.707901
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center