title: Discovery of inter-domain stabilizers-a novel assay system for allosteric akt inhibitors.
abstract: In addition to the catalytically active kinase domain, most kinases feature regulatory domains that govern their activity. Modulating and interfering with these interdomain interactions presents a major opportunity for understanding biological systems and developing novel therapeutics. Therefore, small molecule inhibitors that target these interactions through an allosteric mode of action have high intrinsic selectivity, as these interactions are often unique to a single kinase or kinase family. Here we report the development of iFLiK (interface-Fluorescent Labels in Kinases), a fluorescence-based assay that can monitor such interdomain interactions. Using iFLiK, we have demonstrated selective detection of allosteric Akt inhibitors that induce an inactive closed conformation unique to Akt. This methodology easily distinguished small molecule allosteric inhibitors from classic ATP-competitive inhibitors. Screening an in-house compound library with iFLiK, we were able to identify novel compounds with a scaffold that has not been previously described for allosteric Akt inhibitors.
authors: Technische Universität Dortmund , Fakultät für Chemie und Chemische Biologie, Otto-Hahn-Strasse 6, 44227 Dortmund, Germany.
authors: Zhizhou
authors: Z
authors: Fang
authors: Jeffrey R
authors: JR
authors: Simard
authors: Dennis
authors: D
authors: Plenker
authors: Hoang D
authors: HD
authors: Nguyen
authors: Trang
authors: T
authors: Phan
authors: Patrik
authors: P
authors: Wolle
authors: Stefan
authors: S
authors: Baumeister
authors: Daniel
authors: D
authors: Rauh
mesh_terms: D000494:Allosteric Regulation; D000495:Allosteric Site; D020134:Catalytic Domain; D045744:Cell Line, Tumor; D055808:Drug Discovery / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008958:Models, Molecular; D011487:Protein Conformation; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D005456:Fluorescent Dyes; D047428:Protein Kinase Inhibitors; D054852:Small Molecule Libraries; D051057:Proto-Oncogene Proteins c-akt