期刊论文详细信息
eLife
Assessing target engagement using proteome-wide solvent shift assays
Jiaming Li1  José Navarrete-Perea1  Jonathan G Van Vranken1  Dylan C Mitchell1  Steven P Gygi1 
[1] Department of Cell Biology, Harvard Medical School, Boston, United States;
关键词: target engagement;    drug discovery;    mass spectrometry;    proteomics;    Human;   
DOI  :  10.7554/eLife.70784
来源: eLife Sciences Publications, Ltd
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【 摘 要 】

Recent advances in mass spectrometry (MS) have enabled quantitative proteomics to become a powerful tool in the field of drug discovery, especially when applied toward proteome-wide target engagement studies. Similar to temperature gradients, increasing concentrations of organic solvents stimulate unfolding and precipitation of the cellular proteome. This property can be influenced by physical association with ligands and other molecules, making individual proteins more or less susceptible to solvent-induced denaturation. Herein, we report the development of proteome-wide solvent shift assays by combining the principles of solvent-induced precipitation (Zhang et al., 2020) with modern quantitative proteomics. Using this approach, we developed solvent proteome profiling (SPP), which is capable of establishing target engagement through analysis of SPP denaturation curves. We readily identified the specific targets of compounds with known mechanisms of action. As a further efficiency boost, we applied the concept of area under the curve analysis to develop solvent proteome integral solubility alteration (solvent-PISA) and demonstrate that this approach can serve as a reliable surrogate for SPP. We propose that by combining SPP with alternative methods, like thermal proteome profiling, it will be possible to increase the absolute number of high-quality melting curves that are attainable by either approach individually, thereby increasing the fraction of the proteome that can be screened for evidence of ligand binding.

【 授权许可】

CC BY   

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