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Chemical proteomics provides a powerful means to gain systems-level insight into the mode of action of small molecules and/or natural products. In contrast to high-throughput screening efforts which only interrogate selected subproteomes such as kinases and often only consider individual domains, the methodology presented herein allows for the determination of the molecular targets of small molecules or drugs in a more relevant physiological setting. As such, the compound of interest is exposed to the entire variety of cellular proteins considering all naturally occurring posttranslational modifications and activation states. Samples prepared according to the procedures described in this protocol are compatible with lysates from cultured cell lines, primary cells, or samples from biopsies. In combination with state-of-the-art mass spectrometry techniques this approach grants access to a comprehensive view of small molecule-target protein interactions.

Original publication

DOI

10.1007/978-1-4939-3341-9_15

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2016

Volume

1394

Pages

211 - 218

Keywords

Chemical proteomics, Drug discovery, Mode of action, Target deconvolution, Animals, Humans, Proteins, Proteome, Proteomics