title: Design, synthesis and characterization of a highly effective inhibitor for analog-sensitive (as) kinases.
abstract: Highly selective, cell-permeable and fast-acting inhibitors of individual kinases are sought-after as tools for studying the cellular function of kinases in real time. A combination of small molecule synthesis and protein mutagenesis, identified a highly potent inhibitor (1-Isopropyl-3-(phenylethynyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine) of a rationally engineered Hog1 serine/threonine kinase (Hog1(T100G)). This inhibitor has been successfully used to study various aspects of Hog1 signaling, including a transient cell cycle arrest and gene expression changes mediated by Hog1 in response to stress. This study also underscores that the general applicability of this approach depends, in part, on the selectivity of the designed the inhibitor with respect to activity versus the engineered and wild type kinases. To explore this specificity in detail, we used a validated chemogenetic assay to assess the effect of this inhibitor on all gene products in yeast in parallel. The results from this screen emphasize the need for caution and for case-by-case assessment when using the Analog-Sensitive Kinase Allele technology to assess the physiological roles of kinases.
authors.affiliation: Medicinal Chemistry, Department of Chemistry, University of Gothenburg, Göteborg, Sweden.
authors.forename: Michael
authors.initials: M
authors.lastname: Klein
authors.forename: Montse
authors.initials: M
authors.lastname: Morillas
authors.forename: Alexandre
authors.initials: A
authors.lastname: Vendrell
authors.forename: Lars
authors.initials: L
authors.lastname: Brive
authors.forename: Marinella
authors.initials: M
authors.lastname: Gebbia
authors.forename: Iain M
authors.initials: IM
authors.lastname: Wallace
authors.forename: Guri
authors.initials: G
authors.lastname: Giaever
authors.forename: Corey
authors.initials: C
authors.lastname: Nislow
authors.forename: Francesc
authors.initials: F
authors.lastname: Posas
authors.forename: Morten
authors.initials: M
authors.lastname: Grøtli
mesh_terms: D002453:Cell Cycle; D015195:Drug Design; D015870:Gene Expression; D020128:Inhibitory Concentration 50; D009682:Magnetic Resonance Spectroscopy; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*
chemical_list: D047428:Protein Kinase Inhibitors