Plant Hormones pp 159-191 | Cite as

Assaying Auxin Receptor Activity Using SPR Assays with F-Box Proteins and Aux/IAA Degrons

  • Mussa Quareshy
  • Veselina Uzunova
  • Justyna M. Prusinska
  • Richard M. NapierEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1497)


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 words

Surface plasmon resonance (SPR) Biacore F-box proteins Auxin binding Compound screen Kinetics 



The authors are grateful for support from BBSRC, UK, Award BB/L009366.


  1. 1.
    Napier RM, David KM, Perrot-Rechenmann C (2002) A short history of auxin-binding proteins. Plant Mol Biol 49:339–348CrossRefPubMedGoogle Scholar
  2. 2.
    Venis MA, Napier RM (1992) Plant hormone receptors: past, present and future. Biochem Soc Trans 20:55–59CrossRefPubMedGoogle Scholar
  3. 3.
    Dharmasiri N, Dharmasiri S, Estelle M (2005) The F-box protein TIR1 is an auxin receptor. Nature 26:441–445CrossRefGoogle Scholar
  4. 4.
    Kepinski S, Leyser O (2005) The Arabidopsis F-box protein TIR1 is an auxin receptor. Nature 26:446–451CrossRefGoogle Scholar
  5. 5.
    Woo EJ, Marshall J, Bauly J, Chen JG, Venis M, Napier RM, Pickersgill RW (2002) Crystal structure of auxin-binding protein 1 in complex with auxin. EMBO J 17:2877–2885CrossRefGoogle Scholar
  6. 6.
    Tan X, Calderon-Villalobos LI, Sharon M, Zheng C, Robinson CV, Estelle M, Zheng M (2007) Mechanism of auxin perception by the TIR1 ubiquitin ligase. Nature 5:640–645CrossRefGoogle Scholar
  7. 7.
    Calderon-Villalobos LI, Lee S, De Oliveira C, Ivetac A, Brandt W, Armitage L, Sheard LB, Tan X, Parry G, Mao H, Zheng N, Napier R, Kepinski S, Estelle M (2012) A combinatorial TIR1/AFB-Aux/IAA co-receptor system for differential sensing of auxin. Nat Chem Biol 1:477–485CrossRefGoogle Scholar
  8. 8.
    Woodward AW, Bartel B (2005) Auxin: regulation, action, and interaction. Ann Bot 95:707–735CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Lee S, Sundaram S, Armitage L, Evans JP, Hawkes T, Kepinski S, Ferro N, Napier RM (2014) Defining binding efficiency and specificity of auxins for SCF(TIR1/AFB)-Aux/IAA co-receptor complex formation. ACS Chem Biol 21:673–682CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Mussa Quareshy
    • 1
  • Veselina Uzunova
    • 1
  • Justyna M. Prusinska
    • 1
  • Richard M. Napier
    • 1
    Email author
  1. 1.School of Life SciencesUniversity of WarwickCoventryUK

Personalised recommendations