title: Thermal proteome profiling monitors ligand interactions with cellular membrane proteins.
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.
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Heidelberg, Germany.
authors.forename: Friedrich B M
authors.initials: FB
authors.lastname: Reinhard
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Heidelberg, Germany.
authors.forename: Dirk
authors.identifier: http://orcid.org/0000-0001-6662-4105
authors.initials: D
authors.lastname: Eberhard
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Heidelberg, Germany.
authors.forename: Thilo
authors.initials: T
authors.lastname: Werner
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Heidelberg, Germany.
authors.forename: Holger
authors.initials: H
authors.lastname: Franken
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Heidelberg, Germany.|Genome Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Dorothee
authors.initials: D
authors.lastname: Childs
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Heidelberg, Germany.
authors.forename: Carola
authors.initials: C
authors.lastname: Doce
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Heidelberg, Germany.
authors.forename: Maria Fälth
authors.initials: MF
authors.lastname: Savitski
authors.affiliation: Genome Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Wolfgang
authors.identifier: http://orcid.org/0000-0002-0474-2218
authors.initials: W
authors.lastname: Huber
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Heidelberg, Germany.
authors.forename: Marcus
authors.identifier: http://orcid.org/0000-0002-8343-8977
authors.initials: M
authors.lastname: Bantscheff
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Heidelberg, Germany.
authors.forename: Mikhail M
authors.initials: MM
authors.lastname: Savitski
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Heidelberg, Germany.
authors.forename: Gerard
authors.initials: G
authors.lastname: Drewes
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000378:metabolism; D018938:Caco-2 Cells; D006358:Hot Temperature; D006801:Humans; D019169:Jurkat Cells; D020014:K562 Cells; D008024:Ligands; D008565:Membrane Proteins / Q000378:metabolism*; D011485:Protein Binding; D055550:Protein Stability; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D054852:Small Molecule Libraries / Q000494:pharmacology; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014638:Vanadates / Q000494:pharmacology
chemical_list: C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D008024:Ligands; D008565:Membrane Proteins; D020543:Proteome; D011948:Receptors, Antigen, T-Cell; D054852:Small Molecule Libraries; C076715:pervanadate; D014638:Vanadates; C499076:ATP1A1 protein, human; D000254:Sodium-Potassium-Exchanging ATPase