Identification of Protein Targets of Bioactive Small Molecules Using Randomly Photomodified Probes

0499970 - ÚOCHB 2019 RIV US eng J - Journal Article
Šimon, Petr - Knedlík, Tomáš - Blažková, Kristýna - Dvořáková, Petra - Březinová, Anna - Kostka, Libor - Šubr, Vladimír - Konvalinka, Jan - Šácha, Pavel
Identification of Protein Targets of Bioactive Small Molecules Using Randomly Photomodified Probes.
ACS Chemical Biology. Roč. 13, č. 12 (2018), s. 3333-3342. ISSN 1554-8929. E-ISSN 1554-8937
R&D Projects: GA ČR(CZ) GA16-02938S; GA MŠMT LO1302; GA MZd(CZ) NV15-31379A
Institutional support: RVO:61388963 ; RVO:61389013
Keywords : glutamate carboxypeptidase II * inhibitor * design
OECD category: Biochemistry and molecular biology; Polymer science (UMCH-V)
Impact factor: 4.374, year: 2018

Identifying protein targets of bioactive small molecules often requires complex, lengthy development of affinity probes. We present a method for stochastic modification of small molecules of interest with a photo-activatable phenyldiazirine linker. The resulting isomeric mixture is conjugated to a hydrophilic copolymer decorated with biotin and a fluorophore. We validated this approach using known inhibitors of several medicinally relevant enzymes. At least a portion of the stochastic derivatives retained their binding to the target, enabling target visualization, isolation, and identification. Moreover, the mix of stochastic probes could be separated into fractions and tested for binding affinity. The structure of the active probe could be determined and the probe resynthesized to improve binding efficiency. Our approach an thus enable rapid target isolation, identification, and visualization, while providing information required for subsequent synthesis of an optimized probe.
Permanent Link: http://hdl.handle.net/11104/0292149