Thermal proteome profiling monitors ligand interactions with cellular membrane proteins

Nat Methods. 2015 Dec;12(12):1129-31. doi: 10.1038/nmeth.3652. Epub 2015 Nov 2.

Abstract

We extended thermal proteome profiling to detect transmembrane protein-small molecule interactions in cultured human cells. When we assessed the effects of detergents on ATP-binding profiles, we observed shifts in denaturation temperature for ATP-binding transmembrane proteins. We also observed cellular thermal shifts in pervanadate-induced T cell-receptor signaling, delineating the membrane target CD45 and components of the downstream pathway, and with drugs affecting the transmembrane transporters ATP1A1 and MDR1.

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B / metabolism
  • Caco-2 Cells
  • Hot Temperature
  • Humans
  • Jurkat Cells
  • K562 Cells
  • Ligands
  • Membrane Proteins / metabolism*
  • Protein Binding
  • Protein Stability
  • Proteome / analysis*
  • Proteome / metabolism
  • Proteomics / methods*
  • Receptors, Antigen, T-Cell / metabolism
  • Small Molecule Libraries / pharmacology
  • Sodium-Potassium-Exchanging ATPase / metabolism
  • Tandem Mass Spectrometry / methods*
  • Vanadates / pharmacology

Substances

  • ABCB1 protein, human
  • ATP Binding Cassette Transporter, Subfamily B
  • Ligands
  • Membrane Proteins
  • Proteome
  • Receptors, Antigen, T-Cell
  • Small Molecule Libraries
  • pervanadate
  • Vanadates
  • ATP1A1 protein, human
  • Sodium-Potassium-Exchanging ATPase