title: Pull-down analysis of interactions among jasmonic acid core signaling proteins.
abstract: Pull-down assays are key tools to test specific protein-protein interactions and have been particularly fruitful in jasmonate signaling research. Here, we describe a standard protocol in which a matrix-bound "bait" protein, expressed in Escherichia coli, pulls down a "prey" protein that is soluble in a protein extract obtained from Arabidopsis thaliana plant tissues. The pulled-down protein can be detected by immunoblot with protein-specific or epitope-specific antibodies. Such a pull-down method was used to reveal interactions among components of the jasmonic acid signaling, including hormone-dependent coreceptor interaction, homodimerization and heterodimerization of JASMONATE ZIM DOMAIN repressors, and interactions among other corepressor components and with transcription factors. Pull-down assays contributed not only to shape this signaling pathway but also to identify the active jasmonate hormone.
authors.affiliation: Departamento de Genética Molecular de Plantas, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
authors.forename: Sandra
authors.initials: S
authors.lastname: Fonseca
authors.forename: Roberto
authors.initials: R
authors.lastname: Solano
mesh_terms: D017360:Arabidopsis; D029681:Arabidopsis Proteins / Q000737:chemistry*; D015153:Blotting, Western; D003517:Cyclopentanes / Q000378:metabolism*; D004926:Escherichia coli; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry; D054883:Oxylipins / Q000378:metabolism*; D010936:Plant Extracts / Q000737:chemistry; D010937:Plant Growth Regulators / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D015398:Signal Transduction
chemical_list: D029681:Arabidopsis Proteins; D003517:Cyclopentanes; D047908:Intracellular Signaling Peptides and Proteins; D054883:Oxylipins; D010936:Plant Extracts; D010937:Plant Growth Regulators; C011006:jasmonic acid