title: Ligand-gated split-kinases.
abstract: The activity of protein kinases are naturally gated by a variety of physiochemical inputs, such as phosphorylation, metal ions, and small molecules. In order to design protein kinases that can be gated by user-defined inputs, we describe a sequence dissimilarity based approach for identifying sites in protein kinases that accommodate 25-residue loop insertion while retaining catalytic activity. We further demonstrate that the successful loop insertion mutants provide guidance for the dissection of protein kinases into two fragments that cannot spontaneously assemble and are thus inactive but can be converted into ligand-gated catalytically active split-protein kinases. We successfully demonstrate the feasibility of this approach with Lyn, Fak, Src, and PKA, which suggests potential generality.
authors.affiliation: Department of Chemistry and Biochemistry, University of Arizona , 1306 East University Boulevard, Tucson, Arizona 85721, United States.
authors.forename: Karla
authors.initials: K
authors.lastname: Camacho-Soto
authors.forename: Javier
authors.initials: J
authors.lastname: Castillo-Montoya
authors.forename: Blake
authors.initials: B
authors.lastname: Tye
authors.forename: Indraneel
authors.initials: I
authors.lastname: Ghosh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000235:genetics; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics*; D016415:Sequence Alignment; D019061:src-Family Kinases / Q000737:chemistry / Q000235:genetics
chemical_list: D011494:Protein Kinases; D051416:Focal Adhesion Protein-Tyrosine Kinases; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases; D017868:Cyclic AMP-Dependent Protein Kinases