Functional analysis of synthetic DELLA domain peptides and bioactive gibberellin assay using surface plasmon resonance technology

Talanta. 2015 Nov 1;144:502-9. doi: 10.1016/j.talanta.2015.06.069. Epub 2015 Jun 30.

Abstract

DELLA proteins and phytohormone gibberellin act together to control convergence point of plant development. A gibberellin-bound nuclear receptor that interacts with the N-terminal domain of DELLA proteins is required for gibberellin induced degradation of DELLA proteins. N-terminal DELLA domain includes two conserved motifs: DELLA and VHYNP. However, their respective functions remain unclear. Meanwhile, the identification and detection of several bioactive gibberellins from the more than 100 gibberellin metabolites are overwhelmingly difficult for their similar structures. Using in vitro biochemical approach, our work demonstrates for the first time that the synthetic GAI N-terminal DELLA domain peptides have similar bioactive function as the expressed protein to interact with AtGID1a receptor. Furthermore, our results reveal that DELLA motif is vitally important region and DELLA segment is essentially required region to recognize AtGID1a receptor. Finally, based on bioactive GA-dependent of the interaction between AtGID1a and DELLA protein, we generated a new method that could identify and detect bioactive GAs accurately and rapidly with surface plasmon resonance assays.

Keywords: Bioactive gibberellin; DELLA region; Gibberellin receptor; Qualitative and quantitative analysis; SPR; Synthetic peptide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis Proteins / chemistry*
  • Arabidopsis Proteins / isolation & purification
  • Biotin / chemistry
  • Gibberellins / analysis*
  • Gibberellins / chemistry
  • Glutathione Transferase / chemistry
  • Peptides / chemistry*
  • Protein Structure, Tertiary
  • Receptors, Cell Surface / chemistry*
  • Receptors, Cell Surface / isolation & purification
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Surface Plasmon Resonance

Substances

  • Arabidopsis Proteins
  • GID1a protein, Arabidopsis
  • Gibberellins
  • Peptides
  • Receptors, Cell Surface
  • Recombinant Proteins
  • Biotin
  • Glutathione Transferase