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Updated: Aug 9, 2026

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach
Published on: February 21, 2019
Proteomic Profiling of Protein Kinase Inhibitor Targets by Mass Spectrometry
Martin Golkowski1, Dustin J Maly2, Shao-En Ong3
1Departments of Chemistry and Biochemistry, University of Washington, 1959 NE Pacific Street, Seattle, WA, 98195-7280, USA.
Abstract:
Identifying cellular targets of bioactive small molecules from large-scale screening campaigns can be a significant bottleneck in developing novel therapeutics. Our rapid small-molecule target profiling protocol combines affinity enrichment and SILAC for proteomic identification of small molecule-protein interactions. Selective interactions are easily discernable from nonspecific protein binding by quantitative ratios. Using kinase inhibitors as an example, we provide an optimized protocol featuring on-bead protein digestion and single nano-flow liquid chromatographic-mass spectrometric (LC-MS) analyses, consequently increasing analytical throughput and sensitivity over gel-based sample preparation methods for rapid profiling of kinase inhibitor targets.
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