Noncompetitive Allosteric Antagonism of Death Receptor 5 by a Synthetic Affibody Ligand

Nagamani Vunnam, Sophia Szymonski, Petra Hirsova, Gregory J. Gores, Jonathan N. Sachs, Benjamin J. Hackel

Research output: Contribution to journalArticlepeer-review

Abstract

Fatty acid-induced upregulation of death receptor 5 (DR5) and its cognate ligand, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), promotes hepatocyte lipoapoptosis, which is a key mechanism in the progression of fatty liver disease. Accordingly, inhibition of DR5 signaling represents an attractive strategy for treating fatty liver disease. Ligand competition strategies are prevalent in tumor necrosis factor receptor antagonism, but recent studies have suggested that noncompetitive inhibition through perturbation of the receptor conformation may be a compelling alternative. To this end, we used yeast display and a designed combinatorial library to identify a synthetic 58-amino acid affibody ligand that specifically binds DR5. Biophysical and biochemical studies show that the affibody neither blocks TRAIL binding nor prevents the receptor-receptor interaction. Live-cell fluorescence lifetime measurements indicate that the affibody induces a conformational change in transmembrane dimers of DR5 and favors an inactive state of the receptor. The affibody inhibits apoptosis in TRAIL-treated Huh-7 cells, an in vitro model of fatty liver disease. Thus, this lead affibody serves as a potential drug candidate, with a unique mechanism of action, for fatty liver disease.

Original languageEnglish (US)
Pages (from-to)3856-3868
Number of pages13
JournalBiochemistry
Volume59
Issue number40
DOIs
StatePublished - Oct 13 2020

ASJC Scopus subject areas

  • Biochemistry

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