Allosteric Inhibition of Bcr-Abl Kinase by High Affinity Monobody Inhibitors Directed to the Src Homology 2 (SH2)-Kinase Interface
| Autor: | Shohei Koide, Allan Joaquim Lamontanara, John Wojcik, Oliver Hantschel, Louesa R. Akin, Grzegorz J. Grabe, Barbara Gerig, Akiko Koide |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Biochemistry & Molecular Biology Allosteric regulation Fusion Proteins bcr-abl Biology Mitogen-activated protein kinase kinase SH2 domain Biochemistry src Homology Domains Antibodies Monoclonal Murine-Derived 03 medical and health sciences Allosteric Regulation Cell Line Tumor hemic and lymphatic diseases Humans Kinase activity Molecular Biology Kinase 11 Medical And Health Sciences Cell Biology 06 Biological Sciences 3. Good health Monobody 030104 developmental biology Protein Structure and Folding Cyclin-dependent kinase 9 03 Chemical Sciences Proto-oncogene tyrosine-protein kinase Src |
| Zdroj: | Journal of Biological Chemistry. 291:8836-8847 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m115.707901 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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