title: In vitro translation-based protein kinase substrate identification.
abstract: Identification of a particular protein as a physiological substrate towards kinases of interest is an extremely complex process. Under physiological conditions kinases and their putative substrates are in low abundance, and production of active eukaryotic kinases with standard overexpressing methods is an arduous task. Herein, we describe a cell-free in vitro protein translation procedure combined with fluorescent phosphoprotein staining as a simple and rapid method for identification of putative kinase substrates.
authors.affiliation: Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary.
authors.forename: Szilvia K
authors.initials: SK
authors.lastname: Nagy
authors.forename: Tamás
authors.initials: T
authors.lastname: Mészáros
mesh_terms: D002474:Cell-Free System; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D010750:Phosphoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D059748:Proteolysis; D013194:Staining and Labeling; D013379:Substrate Specificity; D014158:Transcription, Genetic
chemical_list: D018076:DNA, Complementary; D010750:Phosphoproteins; D011494:Protein Kinases