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In-depth characterization of Staurosporine induced proteome thermal stability changes

Chernobrovkin, A. L.; Lengqvist, J.; Cacares Korner, C.; Amadio, D.; Friman, T.; Molina, D. M.

2020-03-15 molecular biology
10.1101/2020.03.13.990606 bioRxiv
Show abstract

Cellular thermal shift assay (CETSA) coupled with high-resolution mass-spectrometry (MS) has proven to be indispensable tool to track thermal stability changes in cellular proteins caused by various external or internal perturbations. One of the major applications of CETSA MS is still binding profile characterization of small molecule drugs. Before applying the method for characterization of novel compound it is crucially important to understand the limitations, sensitivity and ways to improve the throughput of the different method implementations. Here we present deep comprehensive profiling of Staurosporine induced proteome thermal stability alteration utilizing different experiment layouts. By applying unbiased straightforward sample preparation and data analysis approaches we were able to compare and benchmark different experiment layouts for detecting compound-induced protein stability changes in CETSA MS experiments.

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