Thermal proteome profiling for interrogating protein interactions

• Published in: Molecular systems biology Vol. 16; no. 3; pp. e9232 - n/a
• Main Authors: Mateus, André, Kurzawa, Nils, Becher, Isabelle, Sridharan, Sindhuja, Helm, Dominic, Stein, Frank, Typas, Athanasios, Savitski, Mikhail M
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.03.2020; EMBO Press; John Wiley and Sons Inc; Springer Nature
• Subjects: Antibiotics; Biological activity; Biophysical Phenomena; Cell cycle; Data analysis; drug discovery; Drugs; EMBO22; EMBO56; Experiments; Heat; Humans; Kinases; Mass Spectrometry; Mass spectroscopy; Metabolites; Nucleic acids; Protein Binding; protein complexes; Protein interaction; Protein Interaction Maps; Protein Stability; Proteins; Proteins - chemistry; Proteins - metabolism; Proteomes; Proteomics; Proteomics - methods; Review; Reviews; Scientific imaging; Target recognition; thermal proteome profiling; Thermal stability; Thermodynamics; protein complexes; drug discovery; proteomics; metabolites; thermal proteome profiling
• ISSN: 1744-4292; 1744-4292
• DOI: 10.15252/msb.20199232

Thermal proteome profiling (TPP) is based on the principle that, when subjected to heat, proteins denature and become insoluble. Proteins can change their thermal stability upon interactions with small molecules (such as drugs or metabolites), nucleic acids or other proteins, or upon post‐translational modifications. TPP uses multiplexed quantitative mass spectrometry‐based proteomics to monitor the melting profile of thousands of expressed proteins. Importantly, this approach can be performed in vitro , in situ , or in vivo . It has been successfully applied to identify targets and off‐targets of drugs, or to study protein–metabolite and protein–protein interactions. Therefore, TPP provides a unique insight into protein state and interactions in their native context and at a proteome‐wide level, allowing to study basic biological processes and their underlying mechanisms. Graphical Abstract This tutorial explains the principles of thermal proteome profiling (TPP) and analyzes the different steps of a TPP experiment. It reviews the recent developments and current applications of this methodology, and provides an outlook of possible new applications.