Reversible protein phosphorylation cascades are the dominant means by which cells process extracellular information and convert these signals into complex phenotypic responses such as growth, differentiation, adhesion and motility. The proper spatial and temporal control of intracellular phosphorylation events by protein kinases is essential for normal cellular function. Indeed, mis-regulation of protein kinase activity is a cause or consequence of a number of diseases, including cancer, diabetes and chronic inflammation. For this reason, protein kinases are one of the most highly pursued targets for the development of new therapeutics by the pharmaceutical industry. Despite widespread interest in the determination of kinase cellular function and in the development of kinase-targeted pharmacological agents, most members of this diverse enzyme family remain poorly characterized. A major reason for this shortcoming is that there remains a paucity of techniques for the global analysis of protein kinases in complex biological mixtures. The overall goal of the research in this proposal is to develop a general set of molecular tools that will allow the functional state of protein kinases to be profiled in cell lysates and living cells.
The Specific Aims of the proposed research are: (1) To develop a panel of active site-directed, photo-affinity probes that allow global profiling of the kinase superfamily. (2) To develop a general set of assays for the analysis of probes developed in Specific Aim 1 and to use these assays to determine the selectivity of several clinically relevant kinase inhibitors. (3) To develop a general set of molecular probes that selectively bind to either the active or inactive conformation of protein kinases. If successful, these tools will provide a general method for determining the cellular targets of kinase inhibitors in physiologically relevant environments and for identifying aberrant kinase activities in human disease.

Public Health Relevance

The mis-regulation of protein kinase activity is a cause or consequence of a number of diseases, including cancer, diabetes and chronic inflammation, which has made members of this diverse enzyme family targets for the development of new therapeutics. The goal of our proposed research is to develop new chemical tools for the molecular analysis of protein kinases in cellular lysates and in living cells. These reagents should allow us to gain a greater understanding of kinase function and aid in the discovery of new targets for the development of therapeutics.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM086858-05
Application #
8325372
Study Section
Synthetic and Biological Chemistry A Study Section (SBCA)
Program Officer
Gerratana, Barbara
Project Start
2008-09-30
Project End
2013-09-14
Budget Start
2012-09-01
Budget End
2013-09-14
Support Year
5
Fiscal Year
2012
Total Cost
$274,743
Indirect Cost
$78,723
Name
University of Washington
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Crowther, Gregory J; Hillesland, Heidi K; Keyloun, Katelyn R et al. (2016) Biochemical Screening of Five Protein Kinases from Plasmodium falciparum against 14,000 Cell-Active Compounds. PLoS One 11:e0149996
Huang, Wenlin; Hulverson, Matthew A; Zhang, Zhongsheng et al. (2016) 5-Aminopyrazole-4-carboxamide analogues are selective inhibitors of Plasmodium falciparum microgametocyte exflagellation and potential malaria transmission blocking agents. Bioorg Med Chem Lett 26:5487-5491
Feldman, Hannah C; Tong, Michael; Wang, Likun et al. (2016) Structural and Functional Analysis of the Allosteric Inhibition of IRE1α with ATP-Competitive Ligands. ACS Chem Biol 11:2195-205
Gower, Carrie M; Thomas, Jason R; Harrington, Edmund et al. (2016) Conversion of a Single Polypharmacological Agent into Selective Bivalent Inhibitors of Intracellular Kinase Activity. ACS Chem Biol 11:121-31
Vidadala, Rama Subba Rao; Rivas, Kasey L; Ojo, Kayode K et al. (2016) Development of an Orally Available and Central Nervous System (CNS) Penetrant Toxoplasma gondii Calcium-Dependent Protein Kinase 1 (TgCDPK1) Inhibitor with Minimal Human Ether-a-go-go-Related Gene (hERG) Activity for the Treatment of Toxoplasmosis. J Med Chem 59:6531-46
Ojo, Kayode K; Dangoudoubiyam, Sriveny; Verma, Shiv K et al. (2016) Selective inhibition of Sarcocystis neurona calcium-dependent protein kinase 1 for equine protozoal myeloencephalitis therapy. Int J Parasitol 46:871-880
Pedroni, Monica J; Vidadala, Rama Subba Rao; Choi, Ryan et al. (2016) Bumped kinase inhibitor prohibits egression in Babesia bovis. Vet Parasitol 215:22-8
Doggett, J Stone; Ojo, Kayode K; Fan, Erkang et al. (2014) Bumped kinase inhibitor 1294 treats established Toxoplasma gondii infection. Antimicrob Agents Chemother 58:3547-9
Gower, Carrie M; Chang, Matthew E K; Maly, Dustin J (2014) Bivalent inhibitors of protein kinases. Crit Rev Biochem Mol Biol 49:102-15
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

Showing the most recent 10 out of 42 publications