@article{3d5db2d6efb1460eac2663921c6fe73c,
title = "Discovery of Protein-Protein Interaction Inhibitors by Integrating Protein Engineering and Chemical Screening Platforms",
abstract = "Maculins et al. describe an approach for identifying small molecule inhibitors of protein-protein interactions that are considered “hard to target.” The team engineered high-affinity variants of one interacting partner that enabled robust chemical screening. Top hits identified using this approach inhibited cellular signaling driven by the endogenous interaction.",
keywords = "LUBAC, NF-κB, cellular screening, high throughput, hit identification, inhibitor, protein engineering, protein-protein interaction, small molecule, ubiquitin",
author = "Timurs Maculins and Javier Garcia-Pardo and Anamarija Skenderovic and Jakob Gebel and Mateusz Putyrski and Andrew Vorobyov and Philipp Busse and Gabor Varga and Maria Kuzikov and Andrea Zaliani and Simin Rahighi and Veronique Schaeffer and Parnham, {Michael J.} and Sidhu, {Sachdev S.} and Andreas Ernst and Volker D{\"o}tsch and Masato Akutsu and Ivan Dikic",
note = "Funding Information: We thank Hadir Marei for comments on the manuscript and useful suggestions on experimental design. We thank Masuda Sader, Meike Hoffmeister, and Elena Veshkova for technical assistance, and Florian Bonn for assistance with IP-MS data analysis. We also acknowledge Katrin Rittinger and Daniel Durocher for reagents. We also thank Aditya Murthy and the department of Cancer Immunology at Genentech for support during final stages of manuscript preparation. This work was supported by the State of Hessen Landes-Offensive zur Entwicklung Wissenschaftlich-?konomischer Exzellenz (LOEWE) Center for Translational Medicine and Pharmacology, TMP, and the Fraunhofer Cluster of Excellence for Immune Mediated Diseases (CIMD), and by grants from Else Kroener Fresenius Stiftung and Dr. Rolf M. Schwiete Stiftung (to I.D.). T.M. and I.D. designed the conceptual framework of the study and experiments. T.M. designed, performed, and analyzed experiments to characterize Ubv-A binding to UBAN and its mechanism of action, validate UBAN: Ubv-A crystal structure, HTS assays, the triage cascade, and hit validation. J.G.-P. studied the in vitro toxicity of hits, and contributed to structural characterization, STD-NMR experiments, analysis, and design of the experiments. A.S. contributed to the characterization of Ubv-A binding to UBAN and its mechanism of action. J.G. performed STD-NMR measurements. M.P. contributed to generation and characterization of UBAN binders. A.V. purified constructs for BLI assays. P.B. contributed to the purification of the NEMO UBAN domain used for structural studies and STD-NMR. G.V. contributed to Ubv-A IP-MS experiments. M.K. contributed to HTS assay development and performed HTS. A.Z. performed chemical analysis and selection of hits. S.R. demonstrated selectivity binding of Ubv-A to UBAN using SPR. V.S. investigated Ubv-A effect on EGFR signaling. M.J.P. contributed to HTS triage design. S.S.S. contributed to phage display design. A.E. contributed to phage display and conceptual study design. V.D. contributed to STD-NMR design. M.A. solved UBAN: Ubv-A crystal structure. T.M. wrote the manuscript with contributions from J.G.-P. A.E. M.A. and I.D. and comments from all authors. 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. Funding Information: We thank Hadir Marei for comments on the manuscript and useful suggestions on experimental design. We thank Masuda Sader, Meike Hoffmeister, and Elena Veshkova for technical assistance, and Florian Bonn for assistance with IP-MS data analysis. We also acknowledge Katrin Rittinger and Daniel Durocher for reagents. We also thank Aditya Murthy and the department of Cancer Immunology at Genentech for support during final stages of manuscript preparation. This work was supported by the State of Hessen Landes-Offensive zur Entwicklung Wissenschaftlich-{\"o}konomischer Exzellenz ( LOEWE ) Center for Translational Medicine and Pharmacology, TMP, and the Fraunhofer Cluster of Excellence for Immune Mediated Diseases (CIMD), and by grants from Else Kroener Fresenius Stiftung and Dr. Rolf M. Schwiete Stiftung (to I.D.). Publisher Copyright: {\textcopyright} 2020 Elsevier Ltd",
year = "2020",
month = nov,
day = "19",
doi = "10.1016/j.chembiol.2020.07.010",
language = "English",
volume = "27",
pages = "1441--1451.e7",
journal = "Cell Chemical Biology",
issn = "2451-9448",
publisher = "Elsevier Inc.",
number = "11",
}