An isothermal shift assay for proteome scale drug-target identification

摘要

Representative vehicle and drug thermal gradient curves indicating the shift in apparent upon drug binding, as measured by TPP (ΔX), and the detection of a shift in the melting curves by measurement of differential protein solubility by iTSA (ΔY). Distribution of Savitski et al . K562 proteome (  = 5985) and staurosporine target data (  = 60) is shown. The first and third quartiles of are delimited by the box, and the median is indicated as the horizontal line within the box. Notches extend to ±1.58 IQR/sqrt(n), where IQR indicates the interquartile range, and whiskers delimit the last data points within 1.5-fold of the IQR. Volcano plot visualization of iTSA staurosporine targets from K562 cell lysates, performed at 52 °C using 20 μM staurosporine. -value = 0.001 is indicated by a solid horizontal line. Histogram of the number of significant staurosporine targets identified by iTSA 52 °C, TPP from Savitski et al. and in house (Ball-Webb) TPP. Targets are categorized as a kinases or ATP-binding protein, or as Other when not annotated as a kinase or ATP-binding protein.