Thermal proteome profiling (TPP) methodology has the potential to characterize proteome-wide stability highlighting key proteins during T. cruzi infection and life stage transition from the invertebrate to the mammalian host. In the present work, T. cruzi epimastigotes and trypomastigotes cell ...
METAL-TPP profiling with another chelator, TPEN, uncovered additional MBPs in proteomes. Collectively, this study developed a robust tool for proteomic discovery of MBPs and provides a rich resource for functional studies of metals in cell biology. This is a preview of subscription content, access...
Deep thermal profiling assigns peptides to proteoforms To systematically measure the melting behavior of proteins in cALL cell lines representing different molecular subtypes, we performed temperature-range TPP8 with eight temperatures per sample and multiplexed two cell lines at a time using TMTpro19,...
Thermal proteome profiling in cell extracts Na-ATP, Mg-ATP, and Na-GTP stocks (300 mM) were prepared in PBS containing 1.5 mM MgCl2 and the pH was adjusted to 7 using NaOH. For TPP-temperature range (TPP-TR) experiments11, two parts of the lysates (both crude and gel filtered),...
To explore the utility of thermal profiling-based methods for analysis of protein sequence variants that result in pronounced cellular fitness defects within the context of the proteome, we have performed mutant TPP analysis (mTPP) in TS mutant yeast using quantitative MS (57, 58, 59). Missense...
The direct detection of drug-protein interactions in living cells is a major challenge in drug discovery research. Recently, we introduced an approach termed thermal proteome profiling (TPP), which enables the monitoring of changes in protein thermal stability across the proteome using quantitative mass...
Mass spectrometry based CEllular Thermal Shift Assay or Thermal Proteome Profiling (TPP) is a recent method that can assess system-wide protein binding to small molecules, metabolites, or nucleic acids by monitoring changes in protein thermal stability19,20. Similar techniques such as stability of pr...
Thermal proteome profiling (TPP) is a novel technique for the identification of active small molecule target proteins based on the principle that protein-ligand binding can increase the thermal stability of proteins. For this purpose, we used the TPP technique in combination with SWATH-DIA mass ...
We have used a mass spectrometry-based proteomic approach to compile an atlas of the thermal stability of 48,000 proteins across 13 species ranging from archaea to humans and covering melting temperatures of 30–90 °C. Protein sequence, composition a
In this project, we detected targets of celecoxib within the nervous system using a label-free TPP (Thermal Proteome Profiling) method. First, proteins of the rat hippocampus were treated with multiple drug concentrations and temperatures. Next, we separated the soluble proteins from the denatured ...