title: Optimal Sox-based fluorescent chemosensor design for serine/threonine protein kinases.
abstract: Fluorescent chemosensors of protein kinase activity provide a continuous, high-throughput sensing format for the study of the roles of these enzymes, which are crucial for regulating cellular function. Specifically, chemosensors using the nonnatural amino acid, Sox, and physiological Mg(2+) levels report phosphorylation with dramatic fluorescence changes that are amenable to real-time and high-throughput analysis. In this article, we report 15 probes for a total of six distinct serine/threonine kinases with large fluorescence increases and good reactivity toward the target kinase. The sensing mechanism is detailed, and the optimal sensing motif is determined. These versatile and powerful sensors provide tools for researchers studying the roles of the targeted kinases in signal transduction, and the design principles provide guidelines for the generation of future fluorescent chemosensors for any serine/threonine kinase.
authors.affiliation: Department of Chemistry, Massachusetts Institute of Technology, Cambridge, 02139, USA.
authors.forename: Melissa D
authors.initials: MD
authors.lastname: Shults
authors.forename: Dora
authors.initials: D
authors.lastname: Carrico-Moniz
authors.forename: Barbara
authors.initials: B
authors.lastname: Imperiali
mesh_terms: D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D007211:Indoles / Q000494:pharmacology; D007700:Kinetics; D008301:Maleimides / Q000494:pharmacology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000138:chemical synthesis / Q000737:chemistry*
chemical_list: D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D007211:Indoles; D008301:Maleimides; D014157:Transcription Factors; C115989:bisindolylmaleimide V; D017346:Protein Serine-Threonine Kinases; C070515:bisindolylmaleimide I; C088060:bisindolylmaleimide