Dysregulation of protein kinase activity has been implicated in a number of diseases, including cancer, diabetes and chronic inflammation. Therefore, protein kinases have emerged as one of the most important drug targets in modern drug discovery. These efforts have resulted in the development of a vast array of small molecule inhibitors that interact with the ATP-binding sites of protein kinases and are able to block their catalytic activities. Interestingly, many of these pharmacological agents exploit the conformational flexibility of the ATP-binding sites of protein kinases by binding to different actie site conformations. For example, the drugs Gleevec (imatinib) and Nexavar (sorafenib) bind to an ATP-binding site conformation of their kinase targets that involves displacement of a catalytically important structural element called the DFG motif. While it has been speculated that certain modes of kinase inhibition lead to more desirable clinical outcomes, there have not been any systematic comparisons at the biochemical and/or cellular level. Our hypothesis is that ATP-competitive inhibitors that stabilize different ATP-binding site conformations of multi-domain protein kinases will have divergent effects on domains and interactions outside of the active site. This leads to the exciting possibility that it may be possible to obtain different phenotypic responses through the inhibition of the same kinase active site by varying the mode of ATP-binding site occupancy. To test this hypothesis, we propose to study how different modes of inhibition affect the SRC family of kinases (SFKs), which are well-characterized, multi- domain tyrosine kinases. SFKs play diverse roles in multiple signaling processes and are important therapeutic targets. Furthermore, it has been demonstrated that beyond their catalytic activities, SFKs also have a number of non-catalytic scaffolding functions that are dependent on their regulatory domains.

Public Health Relevance

This project is focused on characterizing how stabilizing the active sites of SRC-family kinases in specific conformations affect their biochemical and cellular properties. SRC-family kinases are potential therapeutic targets for a number of diseases including cancer and chronic inflammation. These multi-domain enzymes play diverse catalytic and non-catalytic roles in the cell. Exploring how specific active site conformations affect the signaling properties of SRC-family kinases will inform the development of new therapeutics for cancer and inflammation.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM086858-06
Application #
8577462
Study Section
Synthetic and Biological Chemistry A Study Section (SBCA)
Program Officer
Gerratana, Barbara
Project Start
2008-09-30
Project End
2017-08-31
Budget Start
2013-09-15
Budget End
2014-08-31
Support Year
6
Fiscal Year
2013
Total Cost
$303,593
Indirect Cost
$107,093
Name
University of Washington
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Leonard, Stephen E; Register, A C; Krishnamurty, Ratika et al. (2014) Divergent modulation of Src-family kinase regulatory interactions with ATP-competitive inhibitors. ACS Chem Biol 9:1894-905
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Hari, Sanjay B; Merritt, Ethan A; Maly, Dustin J (2014) Conformation-selective ATP-competitive inhibitors control regulatory interactions and noncatalytic functions of mitogen-activated protein kinases. Chem Biol 21:628-35
Register, A C; Leonard, Stephen E; Maly, Dustin J (2014) SH2-catalytic domain linker heterogeneity influences allosteric coupling across the SFK family. Biochemistry 53:6910-23
Vidadala, Rama Subba Rao; Ojo, Kayode K; Johnson, Steven M et al. (2014) Development of potent and selective Plasmodium falciparum calcium-dependent protein kinase 4 (PfCDPK4) inhibitors that block the transmission of malaria to mosquitoes. Eur J Med Chem 74:562-73
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Ojo, Kayode K; Reid, Molly C; Kallur Siddaramaiah, Latha et al. (2014) Neospora caninum calcium-dependent protein kinase 1 is an effective drug target for neosporosis therapy. PLoS One 9:e92929
Golkowski, Martin; Brigham, Jennifer L; Perera, Gayani K et al. (2014) Rapid profiling of protein kinase inhibitors by quantitative proteomics. Medchemcomm 5:363-369

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