title: Measurement of plant cyclin-dependent kinase activity using immunoprecipitation-coupled and affinity purification-based kinase assays and the baculovirus expression system.
abstract: Orderly progression of the eukaryotic cell cycle is governed by a coordinated response to intrinsic and extracellular cues through activation of cyclin-dependent kinases (CDKs). It is therefore important to verify the kinase activity of distinct types of CDKs during the cell cycle. The immunoprecipitation-coupled kinase assay is a useful procedure to evaluate CDK activity in vivo. Although a specific antibody is usually required for immunoprecipitation, transgenic plant cells expressing tag- or marker protein-fused CDKs are also suitable for this purpose. In addition, the baculovirus expression system is a valuable tool for analyzing CDK activity in vitro, because activation of CDKs is regulated by posttranscriptional modification systems that are active in the insect host cells.
authors.affiliation: The International Center for Biotechnology, Osaka University, Suita, Osaka, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Harashima
authors.forename: Masami
authors.initials: M
authors.lastname: Sekine
mesh_terms: D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D002846:Chromatography, Affinity; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016213:Cyclins / Q000378:metabolism; D057075:Enzyme Assays / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D047468:Immunoprecipitation; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014026:Nicotiana / Q000201:enzymology
chemical_list: D016213:Cyclins; D011994:Recombinant Proteins; D018844:Cyclin-Dependent Kinases