Native Mass Spectrometry for the study of PROTAC GNE-987-containing ternary complexes

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Sternicki, Louise M
Nonomiya, Jim
Liu, Miaomiao
Mulvihill, Melinda M
Quinn, Ronald J
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2021
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Abstract

PRO teolysis TA rgeting C himeras (PROTACs) promote the degradation, rather than inhibition, of a drug target as a mechanism for therapeutic treatment. Bifunctional PROTAC molecules allow simultaneous binding of both the target protein and an E3-Ubiquitin ligase, bringing the two proteins into close spatial proximity to allow ubiquitinylation and degradation of the target protein via the cell's endogenous protein degradation pathway. We utilize native MS to study the ternary complexes promoted by the previously reported PROTAC GNE-987 between Brd4 bromodomains 1 and 2, and Von Hippel Lindeau E3-Ubiquitin Ligase. Native MS at high resolution allowed us to measure ternary complex formation as a function of PROTAC concentration to provide a measure of complex affinity and stability, whilst simultaneously measuring other intermediate protein species. Native MS provides a high-throughput, low sample consumption, direct screening method to measure ternary complexes for PROTAC development.

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ChemMedChem

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© 2021 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. This is the peer reviewed version of the following article: Native Mass Spectrometry for the study of PROTAC GNE-987-containing ternary complexes, ChemMedChem, 2021, which has been published in final form at DOI. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving (http://olabout.wiley.com/WileyCDA/Section/id-828039.html)

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Medicinal and biomolecular chemistry

Organic chemistry

Pharmacology and pharmaceutical sciences

GNE-987

PROTAC

Proteolysis Targeting Chimeras

high-throughput screening

native MS

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Sternicki, LM; Nonomiya, J; Liu, M; Mulvihill, MM; Quinn, RJ, Native Mass Spectrometry for the study of PROTAC GNE-987-containing ternary complexes., ChemMedChem, 2021

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