The long-term goal of this project is to develop a robust proteomics approach to identify targets of small-molecule compounds and drugs in live cells. Phenotypic screens are commonly used to identify novel bioactive small-molecule probes and develop new therapeutic approaches. The major bottleneck in forward chemical-genomics screens is target identification and validation of novel small-molecule probes. Strikingly, targets are still unknown for many FDA-approved drugs and small-molecule "tool" compounds. Identifying both the therapeutic targets and "off targets" allows optimization of a compound's selectivity and should decrease potential side effects in the eventual drug. Our proposal describes a novel strategy to use small-molecule inhibitors as in vivo targeting moieties to biotinylate their molecular targets and their protein complexes for proteomic identification. This novel approach is distinct from traditional biochemical approaches and may allow the identification of much weaker affinity binders and higher order protein-protein interactions. As our approach explores spatial relationships in multi-protein complexes when a small-molecule probe is bound to its protein target, it may provide a novel perspective to characterize the small molecule's mechanisms of action.

Public Health Relevance

The discovery of novel therapeutics relies on understanding the mechanisms of actions of small- molecule probes and drugs. Current approaches for target identification rely on protein fragments or extracted cell lysates under non-native conditions. We propose to develop approaches to study protein targets of small-molecule probes within the cell. Such tools will resolve a major bottleneck in drug discovery programs and can accelerate the discovery of new therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21EB018384-01
Application #
8680550
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Korte, Brenda
Project Start
2014-03-01
Project End
2016-02-29
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
1
Fiscal Year
2014
Total Cost
$231,750
Indirect Cost
$81,750
Name
University of Washington
Department
Pharmacology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195