Thermal proteome profiling reveals distinct target selectivity for differentially oxidized oxysterols
Description
The dataset contains excel macro used for the data analysis from Thermal Proteomic Profiling exeriments aimed to identify the protein targets of three A- and B-ring oxidized sterols (cholestane-3β,5α,6β-triol (CT), 7keto-cholesterol (7-KC), 4ß-hydroxycholesterol (4ß-HC) ) and a side-chain oxidized sterol (25-hydroxycholesterol (25-HC)). The excel macros contain the list of the identified proteome in the experiments with related thermal shifts (ΔTm) measured in three independent experiments.
Files
Steps to reproduce
The data were produced applying the protocol from Reckzeh, E. S., et al. (https://doi.org/10.1007/978-1-4939-8891-4_4). Briefly HeLa cells were cultured and lysed with PBS 0.4%NP-40 before incubation for 10 minutes with either oxysterol or DMSO. A gradient temperature ranging from 37 to 67 degree Celsius was applied to the samples and protein soluble fraction was collected for further reduction/alkylation prior tryptic digestion. After TMT labelling, the samples were fractionated with high-pH UHPLC system before mass spectrometric detection with Q-Exactive HF-X (ThermoFisher Scientific). Mass spectrometric raw data were analysed using MaxQuant (version 1.6.2.10). The protein group files from the MaxQuant analysis were processed in the Excel Macro (TPP_Makro_1.0, http:// www.mpi-dortmund.mpg.de/forschung/chemische- biologie/janning). Instruction and explanation of the Excel Macro are described in the first sheet of the files.