Lipid Activity Probes for the Chemical Proteomic Characterization of Protein Targets
EMSL Project ID
47457
Abstract
Lipids including diacyglycerol, phosphatidic acid and the phosphatidylinositol polyphosphates regulate numerous critical biological processes, many of which are aberrant in disease or involved in the production of important biofuels. These lipids often act as site-specific ligands in interactions that enforce membrane-association of protein binding partners. Despite the fact that these signaling lipids control key biological events, many of these roles are poorly understood due to the complexity of the membrane environment and the pathways in which lipids operate. In particular, it remains a significant challenge to discover and characterize proteins that bind and modify lipids. Herein, we describe the development and application of synthetic bifunctional lipid activity probes for the efficient chemical proteomic identification and analysis of proteins that interact with lipids. This approach entails activity-based protein profiling, which involves the collective labeling, purification and identification of target proteins from complex samples such as cell extracts, live cells and organisms. Towards this end, lipid activity probes containing both a photoaffinity tag for covalent labeling of bound proteins as well as a secondary “clickable” handle for subsequent detection or manipulation of labeled proteins have been designed and studied. In initial experiments, these compounds have proven to be effective for the labeling and processing of target proteins based on specific interactions, and ultimately collective identification via high-throughput mass spectrometry-based proteomics. The studies detailed in this user proposal are designed to further advance this technology to study a range of new lipids and investigate the roles of signaling lipids in processes related to disease onset and biofuels production.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2012-10-01
End Date
2014-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members