Structural insights into the disruption of TNF-TNFR1 signalling by small molecules stabilising a distorted TNF

Nat Commun. 2021 Jan 25;12(1):582. doi: 10.1038/s41467-020-20828-3.


Tumour necrosis factor (TNF) is a trimeric protein which signals through two membrane receptors, TNFR1 and TNFR2. Previously, we identified small molecules that inhibit human TNF by stabilising a distorted trimer and reduce the number of receptors bound to TNF from three to two. Here we present a biochemical and structural characterisation of the small molecule-stabilised TNF-TNFR1 complex, providing insights into how a distorted TNF trimer can alter signalling function. We demonstrate that the inhibitors reduce the binding affinity of TNF to the third TNFR1 molecule. In support of this, we show by X-ray crystallography that the inhibitor-bound, distorted, TNF trimer forms a complex with a dimer of TNFR1 molecules. This observation, along with data from a solution-based network assembly assay, leads us to suggest a model for TNF signalling based on TNF-TNFR1 clusters, which are disrupted by small molecule inhibitors.

MeSH terms

  • Algorithms
  • Animals
  • Binding, Competitive / drug effects
  • Humans
  • Models, Molecular
  • Protein Binding / drug effects
  • Protein Conformation / drug effects
  • Protein Multimerization / drug effects*
  • Receptors, Tumor Necrosis Factor, Type I / chemistry*
  • Receptors, Tumor Necrosis Factor, Type I / metabolism
  • Signal Transduction / drug effects*
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology*
  • Tumor Necrosis Factor-alpha / chemistry*
  • Tumor Necrosis Factor-alpha / metabolism


  • Receptors, Tumor Necrosis Factor, Type I
  • Small Molecule Libraries
  • TNF protein, human
  • Tumor Necrosis Factor-alpha