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Noncompetitive allosteric antagonism of death receptor 5 by an affibody discovered via yeast display
Author(s) -
Vunnam Nagamani,
Szymonski Sophia,
Petra Hirsova,
Gores Gregory James,
Hackel Benjamin,
Sachs Jonathan
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.05692
Subject(s) - allosteric regulation , antagonism , receptor , tumor necrosis factor alpha , mechanism of action , chemistry , microbiology and biotechnology , downregulation and upregulation , apoptosis , biochemistry , cancer research , biology , pharmacology , in vitro , immunology , gene
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 is prevalent in tumor necrosis factor receptor antagonism, but recent studies suggest a compelling alternative: non‐competitive inhibition through perturbation of receptor conformation. We used a yeast‐displayed combinatorial library to identify a synthetic affibody 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‐ or palmitate‐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. Support or Funding Information National Institutes of Health Grants R01EB023339, R01DK41876, R35GM131814 and R01EB028274.