Assaying Auxin Receptor Activity Using SPR Assays with F-Box Proteins and Aux/IAA Degrons
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.
Key wordsSurface plasmon resonance (SPR) Biacore F-box proteins Auxin binding Compound screen Kinetics
The authors are grateful for support from BBSRC, UK, Award BB/L009366.