Discovery of Protein-Protein Interaction Inhibitors by Integrating Protein Engineering and Chemical Screening Platforms.
Biological Products
/ chemistry
Cell Line
Cell Survival
/ drug effects
Dose-Response Relationship, Drug
Drug Discovery
Drug Evaluation, Preclinical
Humans
Molecular Structure
NF-kappa B
/ antagonists & inhibitors
Protein Binding
/ drug effects
Protein Engineering
Signal Transduction
/ drug effects
Structure-Activity Relationship
Ubiquitin
/ antagonists & inhibitors
LUBAC
NF-κB
cellular screening
high throughput
hit identification
inhibitor
protein engineering
protein-protein interaction
small molecule
ubiquitin
Journal
Cell chemical biology
ISSN: 2451-9448
Titre abrégé: Cell Chem Biol
Pays: United States
ID NLM: 101676030
Informations de publication
Date de publication:
19 11 2020
19 11 2020
Historique:
received:
21
05
2020
revised:
24
06
2020
accepted:
09
07
2020
pubmed:
30
7
2020
medline:
9
7
2021
entrez:
30
7
2020
Statut:
ppublish
Résumé
Protein-protein interactions (PPIs) govern intracellular life, and identification of PPI inhibitors is challenging. Roadblocks in assay development stemming from weak binding affinities of natural PPIs impede progress in this field. We postulated that enhancing binding affinity of natural PPIs via protein engineering will aid assay development and hit discovery. This proof-of-principle study targets PPI between linear ubiquitin chains and NEMO UBAN domain, which activates NF-κB signaling. Using phage display, we generated ubiquitin variants that bind to the functional UBAN epitope with high affinity, act as competitive inhibitors, and structurally maintain the existing PPI interface. When utilized in assay development, variants enable generation of robust cell-based assays for chemical screening. Top compounds identified using this approach directly bind to UBAN and dampen NF-κB signaling. This study illustrates advantages of integrating protein engineering and chemical screening in hit identification, a development that we anticipate will have wide application in drug discovery.
Identifiants
pubmed: 32726587
pii: S2451-9456(20)30283-X
doi: 10.1016/j.chembiol.2020.07.010
pii:
doi:
Substances chimiques
Biological Products
0
NF-kappa B
0
Ubiquitin
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1441-1451.e7Informations de copyright
Copyright © 2020 Elsevier Ltd. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Interests T.M., A.E., M.P., M.K., and M.J.P. co-author a patent application for a part of this work (EP18191813.7 application number). T.M. is a current employee of Genentech. J.G,-P., A.V., M.K., A.Z., M.J.P., and I.D. are current employees of Fraunhofer Institutes. A.S. is a current employee of Pliva Croatia. M.P. is a current employee of Bio-Rad.