Identification of a covalent binder to the oncoprotein gankyrin using a NIR-Based OBOC screening method.

Autor:	Maresh ME; Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University West Lafayette Indiana 47907 USA dtrader@purdue.edu., Zerfas BL; Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University West Lafayette Indiana 47907 USA dtrader@purdue.edu., Wuthrich BS; Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University West Lafayette Indiana 47907 USA dtrader@purdue.edu., Trader DJ; Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University West Lafayette Indiana 47907 USA dtrader@purdue.edu.
Jazyk:	angličtina
Zdroj:	RSC advances [RSC Adv] 2021 Apr 01; Vol. 11 (21), pp. 12794-12801. Date of Electronic Publication: 2021 Apr 01 (Print Publication: 2021).
DOI:	10.1039/d0ra10976b
Abstrakt:	Despite huge advancements in the process of synthesizing small molecules as part of one-bead-one-compound (OBOC) libraries, progress lags in the ability to screen these libraries against proteins of interest. Recently, we developed a method to screen OBOC libraries in which a target protein is labeled with a near-infrared (NIR) range fluorophore. The labeled protein incubates with beads of a library in a 96-well plate, then the plate is imaged for fluorescence. Fluorescence intensities produced by the labeled protein binding the bead can be quantitated and provide a basis to rank hits. Here, we present an application of this technique by screening the oncoprotein gankyrin against a 343-member peptoid library. The library was composed of four positions occupied by one of seven amines. In the third position, an amine that facilitates covalent binding via a sulfonyl fluoride moiety was incorporated. After screening for gankyrin binders twice, ten structures showed overlap in the types of amines present at each position. These initial hits were validated with an in-gel fluorescence assay in which the labeled ligands covalently interacted with purified gankyrin. Excitingly, one peptoid was validated from this analysis. This hit was also shown to bind gankyrin in the presence of HEK 293T lysate. Results from this study demonstrate successful use of our screening method to quickly identify quality binders to a target protein of interest. Competing Interests: Prof. Trader is a scientific advisor and shareholder of Booster Therapeutics GmbH. Other authors declare no conflicts. (This journal is © The Royal Society of Chemistry.)
Databáze:	MEDLINE
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