Thermal proteome profiling reveals distinct target selectivity for differentially oxidized oxysterols

Published: 20 June 2022| Version 2 | DOI: 10.17632/r26nzs66mn.2
Contributors:
Cecilia Rossetti,

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.

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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.

Institutions

Danmarks Tekniske Universitet

Categories

Cell Biology, Proteomics, Interactome

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