4.4 Selections of DNA-Encoded Libraries to Protein Targets within Living Cells
Book
Editors: Scheuermann, J. ; Li, Y.
Title: DNA-Encoded Libraries
Print ISBN: 9783132455221; Online ISBN: 9783132437357; Book DOI: 10.1055/b000000342
1st edition © 2024. Thieme. All rights reserved.
Georg Thieme Verlag KG, Stuttgart
Subjects: Organic Chemistry
Science of Synthesis Reference Libraries
Parent publication
Title: Science of Synthesis
DOI: 10.1055/b-00000101
Series Editors: Fürstner, A. (Editor-in-Chief); Carreira, E. M.; Faul, M.; Kobayashi, S.; Koch, G.; Molander, G. A.; Nevado, C.; Trost, B. M.; You, S.-L.
Type: Multivolume Edition
Abstract
Selections of DNA-encoded libraries (DELs) have been largely limited to biochemically pure proteins that are immobilized on solid supports. This excludes many targets that cannot be purified or reconstituted biochemically in a fully active form. This chapter describes a method for the selection of DEL ligands against proteins within live cells. The approach relies on appending a cyclic cell-penetrating peptide to the DNA construct to deliver DELs across the cell membrane into the cytoplasm of live cells. Once inside the cell, affinity crosslinking is used to covalently trap protein–ligand interactions. The DEL ligands are isolated by subsequent purification of the target protein. This approach has demonstrated potential in selecting focused libraries against cytosolic targets that are challenging to express and purify.
Key words
DNA-encoded libraries - crosslinking - cyclic cell-penetrating peptides - drug discovery - selections- 2 Cai B, Kim D, Akhand S, Sun Y, Cassell RJ, Alpsoy A, Dykhuizen EC, Van Rijn RM, Wendt MK, Krusemark CJ. J. Am. Chem. Soc. 2019; 141: 17057
- 6 Qian Z, Martyna A, Hard RL, Wang J, Appiah-Kubi G, Coss C, Phelps MA, Rossman JS, Pei D. Biochemistry 2016; 55: 2601
- 8 Petersen LK, Christensen AB, Andersen J, Folkesson CG, Kristensen O, Andersen C, Alzu A, Sløk FA, Blakskjær P, Madsen D, Azevedo C, Micco I, Hansen NJV. J. Am. Chem. Soc. 2021; 143: 2751
- 12 Denton KE, Wang S, Gignac MC, Milosevich N, Hof F, Dykhuizen EC, Krusemark CJ. SLAS Discovery 2018; 23: 417
- 13 Simhadri C, Daze KD, Douglas SF, Quon TTH, Dev A, Gignac MC, Peng F, Heller M, Boulanger MJ, Wulff JE, Hof F. J. Med. Chem. 2014; 57: 2874