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, metabolism); Signal Transduction (drug effects); Small Molecule Libraries (chemistry, pharmacology); Tumor Necrosis Factor-alpha (chemistry, metabolism)
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.
Digital Object Identifier (DOI)
Published in Nature Communications, Volume 12, Issue 582, 2021, pages 582-.
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McMillan, D., Martinez-Fleites, C., Porter, J., Fox, D., Davis, R., Mori, P., Ceska, T., Carrington, B., Lawson, A., Bourne, T., & O’Connell, J. (2021). Structural insights into the disruption of TNF-TNFR1 signalling by small molecules stabilising a distorted TNF. Nature Communications, 12(1). https://doi.org/10.1038/s41467-020-20828-3