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A chemoproteomic platform for selective deubiquitinase inhibitor discovery

ABSTRACT: Here we describe our work on the development of an ABPP screening platform for cysteine reactive covalent fragments against deubiquitinating enzymes (DUBs) on 96 well plate format. Within this targeted workflow, we profile cysteine reactive fragments by competition with biotinylated ubiquitin probe in cell lysates and employ label free data independent acquisition (DIA) MS method. The platform was used to screen a library of 138 Cys-reactive covalent fragments against DUBome in HEK293T cell lysate, identifying functionally relevant hit fragments for numerous DUBs and demonstrating the utility of the approach in expanding the liganded proteome.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Aini Vuorinen

LAB HEAD: Jacob Bush

PROVIDER: PXD037152 | Pride | 2024-03-18

REPOSITORIES: Pride

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A chemoproteomic platform for selective deubiquitinase inhibitor discovery

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