title: Assaying Auxin Receptor Activity Using SPR Assays with F-Box Proteins and Aux/IAA Degrons.
abstract: The identification of TIR1 as an auxin receptor combined with advanced biophysical instrumentation has led to the development of real-time activity assays for auxins. Traditionally, molecules have been assessed for auxinic activity using bioassays, and agrochemical compound discovery continues to be based on "spray and pray" technologies. Here, we describe the methodology behind an SPR-based assay that uses TIR1 and related F-box proteins with surface plasmon resonance spectrometry for rapid compound screening. In addition, methods for collecting kinetic binding data and data processing are given so that they may support programs for rational design of novel auxin ligands.
authors.affiliation: School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.
authors.forename: Mussa
authors.initials: M
authors.lastname: Quareshy
authors.affiliation: School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.
authors.forename: Veselina
authors.initials: V
authors.lastname: Uzunova
authors.affiliation: School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.
authors.forename: Justyna M
authors.initials: JM
authors.lastname: Prusinska
authors.affiliation: School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK. Richard.Napier@warwick.ac.uk.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Napier
mesh_terms: D017360:Arabidopsis / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000378:metabolism*; D001681:Biological Assay / Q000379:methods; D044783:F-Box Proteins / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D007210:Indoleacetic Acids / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D010940:Plant Proteins / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology
chemical_list: D029681:Arabidopsis Proteins; D044783:F-Box Proteins; D007210:Indoleacetic Acids; D008024:Ligands; D010940:Plant Proteins; D011956:Receptors, Cell Surface; C501405:TIR1 protein, Arabidopsis; C046131:auxin receptor, plant
keywords: Auxin binding; Biacore; Compound screen; F-box proteins; Kinetics; Surface plasmon resonance (SPR)