title: Abscisic acid signaling: thermal stability shift assays as tool to analyze hormone perception and signal transduction.
abstract: Abscisic acid (ABA) is a plant hormone that plays important roles in growth and development. ABA is also the central regulator to protect plants against abiotic stresses, such as drought, high salinity, and adverse temperatures, and ABA signaling is therefore a promising biotechnological target for the generation of crops with increased stress resistance. Recently, a core signal transduction pathway has been established, in which ABA receptors, type 2C protein phosphatases, and AMPK-related protein kinases control the regulation of transcription factors, ion channels, and enzymes. Here we use a simple protein thermal stability shift assay to independently validate key aspects of this pathway and to demonstrate the usefulness of this technique to detect and characterize very weak (Kd ≥ 50 µM) interactions between receptors and physiological and synthetic agonists, to determine and analyze protein-protein interactions, and to screen small molecule inhibitors.
authors.affiliation: Laboratory of Structural Sciences, Van Andel Research Institute, NE, Grand Rapids, Michigan, United States of America.
authors.forename: Fen-Fen
authors.initials: FF
authors.lastname: Soon
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Suino-Powell
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.forename: Eu-Leong
authors.initials: EL
authors.lastname: Yong
authors.forename: H Eric
authors.initials: HE
authors.lastname: Xu
authors.forename: Karsten
authors.initials: K
authors.lastname: Melcher
mesh_terms: D000040:Abscisic Acid / Q000378:metabolism*; D017360:Arabidopsis / Q000737:chemistry / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000378:metabolism*; D010937:Plant Growth Regulators / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism; D055550:Protein Stability; D011956:Receptors, Cell Surface; D015398:Signal Transduction; D013696:Temperature
chemical_list: D029681:Arabidopsis Proteins; C569327:PYL2 protein, Arabidopsis; D010937:Plant Growth Regulators; D011956:Receptors, Cell Surface; D000040:Abscisic Acid; D011494:Protein Kinases; C472139:OST1 protein, Arabidopsis