title: Tracking cancer drugs in living cells by thermal profiling of the proteome.
abstract: The thermal stability of proteins can be used to assess ligand binding in living cells. We have generalized this concept by determining the thermal profiles of more than 7000 proteins in human cells by means of mass spectrometry. Monitoring the effects of small-molecule ligands on the profiles delineated more than 50 targets for the kinase inhibitor staurosporine. We identified the heme biosynthesis enzyme ferrochelatase as a target of kinase inhibitors and suggest that its inhibition causes the phototoxicity observed with vemurafenib and alectinib. Thermal shifts were also observed for downstream effectors of drug treatment. In live cells, dasatinib induced shifts in BCR-ABL pathway proteins, including CRK/CRKL. Thermal proteome profiling provides an unbiased measure of drug-target engagement and facilitates identification of markers for drug efficacy and toxicity.
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Meyerhofstrasse 1, Heidelberg, Germany. mikhail.m.savitski@gsk.com marcus.x.bantscheff@gsk.com gerard.c.drewes@gsk.com.
authors.forename: Mikhail M
authors.initials: MM
authors.lastname: Savitski
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Friedrich B M
authors.initials: FB
authors.lastname: Reinhard
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Holger
authors.initials: H
authors.lastname: Franken
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Thilo
authors.initials: T
authors.lastname: Werner
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Maria Fälth
authors.initials: MF
authors.lastname: Savitski
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Meyerhofstrasse 1, Heidelberg, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Eberhard
authors.affiliation: Division of Biophysics, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Daniel
authors.initials: D
authors.lastname: Martinez Molina
authors.affiliation: Division of Biophysics, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Rozbeh
authors.initials: R
authors.lastname: Jafari
authors.affiliation: Division of Biophysics, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Rebecca Bakszt
authors.initials: RB
authors.lastname: Dovega
authors.affiliation: Department of Proteomics and Bioanalytics, Technische Universität München, Emil Erlenmeyer Forum 5, Freising, Germany. German Cancer Consortium, German Cancer Research Center, Heidelberg, Germany.
authors.forename: Susan
authors.initials: S
authors.lastname: Klaeger
authors.affiliation: Department of Proteomics and Bioanalytics, Technische Universität München, Emil Erlenmeyer Forum 5, Freising, Germany. German Cancer Consortium, German Cancer Research Center, Heidelberg, Germany.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Kuster
authors.affiliation: Division of Biophysics, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden. Centre for Biomedical Structural Biology, Nanyang Technological University, Singapore.
authors.forename: Pär
authors.initials: P
authors.lastname: Nordlund
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Meyerhofstrasse 1, Heidelberg, Germany. mikhail.m.savitski@gsk.com marcus.x.bantscheff@gsk.com gerard.c.drewes@gsk.com.
authors.forename: Marcus
authors.initials: M
authors.lastname: Bantscheff
authors.affiliation: Cellzome GmbH, Molecular Discovery Research, GlaxoSmithKline, Meyerhofstrasse 1, Heidelberg, Germany. mikhail.m.savitski@gsk.com marcus.x.bantscheff@gsk.com gerard.c.drewes@gsk.com.
authors.forename: Gerard
authors.initials: G
authors.lastname: Drewes
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology*; D006358:Hot Temperature; D006801:Humans; D020014:K562 Cells; D008024:Ligands; D011485:Protein Binding; D011489:Protein Denaturation; D055550:Protein Stability; D020543:Proteome / Q000187:drug effects*; D040901:Proteomics / Q000379:methods*
chemical_list: D000970:Antineoplastic Agents; D008024:Ligands; D020543:Proteome; D000251:Adenosine Triphosphatases