title: A Multiplexed NMR-Reporter Approach to Measure Cellular Kinase and Phosphatase Activities in Real-Time.
abstract: Cell signaling is governed by dynamic changes in kinase and phosphatase activities, which are difficult to assess with discontinuous readout methods. Here, we introduce an NMR-based reporter approach to directly identify active kinases and phosphatases in complex physiological environments such as cell lysates and to measure their individual activities in a semicontinuous fashion. Multiplexed NMR profiling of reporter phosphorylation states provides unique advantages for kinase inhibitor studies and reveals reversible modulations of cellular enzyme activities under different metabolic conditions.
authors.forename: Rossukon
authors.initials: R
authors.lastname: Thongwichian
authors.forename: Jonas
authors.initials: J
authors.lastname: Kosten
authors.affiliation: ‡Max Delbrück Center for Molecular Medicine (MDC Berlin), Robert Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Uwe
authors.initials: U
authors.lastname: Benary
authors.forename: Honor May
authors.initials: HM
authors.lastname: Rose
authors.forename: Marchel
authors.initials: M
authors.lastname: Stuiver
authors.forename: Francois-Xavier
authors.initials: FX
authors.lastname: Theillet
authors.forename: Alexander
authors.initials: A
authors.lastname: Dose
authors.affiliation: ∥European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69123 Heidelberg, Germany.
authors.forename: Birgit
authors.initials: B
authors.lastname: Koch
authors.affiliation: ∥European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69123 Heidelberg, Germany.
authors.forename: Hideki
authors.initials: H
authors.lastname: Yokoyama
authors.forename: Dirk
authors.initials: D
authors.lastname: Schwarzer
authors.affiliation: ‡Max Delbrück Center for Molecular Medicine (MDC Berlin), Robert Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Wolf
authors.forename: Philipp
authors.initials: P
authors.lastname: Selenko
mesh_terms: D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D020014:K562 Cells; D019906:Nuclear Magnetic Resonance, Biomolecular; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D010770:Phosphotransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013997:Time Factors
chemical_list: D047428:Protein Kinase Inhibitors; D010770:Phosphotransferases; D010744:Phosphoric Monoester Hydrolases