Interfering with the high-affinity interaction between wheat amylase trypsin inhibitor CM3 and toll-like receptor 4: in silico and biosensor-based studies

Sci Rep. 2017 Oct 13;7(1):13169. doi: 10.1038/s41598-017-13709-1.


Wheat amylase/trypsin bi-functional inhibitors (ATIs) are protein stimulators of innate immune response, with a recently established role in promoting both gastrointestinal and extra-gastrointestinal inflammatory syndromes. These proteins have been reported to trigger downstream intestinal inflammation upon activation of TLR4, a member of the Toll-like family of proteins that activates signalling pathways and induces the expression of immune and pro-inflammatory genes. In this study, we demonstrated the ability of ATI to directly interact with TLR4 with nanomolar affinity, and we kinetically and structurally characterized the interaction between these macromolecules by means of a concerted approach based on surface plasmon resonance binding analyses and computational studies. On the strength of these results, we designed an oligopeptide capable of preventing the formation of the complex between ATI and the receptor.

MeSH terms

  • Biosensing Techniques*
  • Plant Proteins / metabolism*
  • Protein Binding
  • Surface Plasmon Resonance
  • Toll-Like Receptor 4 / metabolism*
  • Triticum / metabolism*
  • Trypsin Inhibitors / metabolism*


  • Plant Proteins
  • Toll-Like Receptor 4
  • Trypsin Inhibitors