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
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ATP Binding Cassette Transporter, Subfamily B / metabolism
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Caco-2 Cells
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Hot Temperature
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Humans
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Jurkat Cells
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K562 Cells
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Ligands
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Membrane Proteins / metabolism*
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Protein Binding
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Protein Stability
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Proteome / analysis*
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Proteome / metabolism
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Proteomics / methods*
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Receptors, Antigen, T-Cell / metabolism
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Small Molecule Libraries / pharmacology
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Sodium-Potassium-Exchanging ATPase / metabolism
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Tandem Mass Spectrometry / methods*
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Vanadates / pharmacology
Substances
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ABCB1 protein, human
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ATP Binding Cassette Transporter, Subfamily B
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Ligands
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Membrane Proteins
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Proteome
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Receptors, Antigen, T-Cell
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Small Molecule Libraries
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pervanadate
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Vanadates
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ATP1A1 protein, human
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Sodium-Potassium-Exchanging ATPase