title: Sensitizing plant protein kinases to specific inhibition by ATP-competitive molecules.
abstract: The highly conserved nature of the protein kinase catalytic domain and the low permeability of plant cell membranes pose a challenge to the development of specific inhibitors that target individual protein kinases in vivo. Here, we describe a chemical-genetic approach to specifically sensitize individual plant kinases to cell-permeable small molecules that do not inhibit wild-type kinases. In this approach, a single amino-acid substitution is introduced in the ATP-binding site of the enzyme enabling specific binding of ATP-competitive molecules. Cell-permeable molecules can then be used to specifically target the sensitized allele in transgenic Arabidopsis thaliana plants that do not express the wild-type form of the kinase. This strategy provides a useful tool for the functional characterization of protein kinases in planta and for the dissection of the signaling pathways in which they are involved.
authors.affiliation: Department of Plant Sciences, Tel-Aviv University, Tel-Aviv, Israel.
authors.forename: Dor
authors.initials: D
authors.lastname: Salomon
authors.forename: Chao
authors.initials: C
authors.lastname: Zhang
authors.forename: Kevan M
authors.initials: KM
authors.lastname: Shokat
authors.forename: Guido
authors.initials: G
authors.lastname: Sessa
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D001667:Binding, Competitive / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D057075:Enzyme Assays; D018506:Gene Expression Regulation, Plant; D006720:Homozygote; D009154:Mutation / Q000235:genetics; D010944:Plants / Q000201:enzymology*; D030821:Plants, Genetically Modified; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011494:Protein Kinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D047428:Protein Kinase Inhibitors; D011993:Recombinant Fusion Proteins; D000255:Adenosine Triphosphate; D011494:Protein Kinases