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CellEKT: A chemical proteomics assay to profile cellular target engagement of kinase inhibitors

ABSTRACT: Activity-based protein profiling is a powerful tool to study target engagement in complex biological environments. XO44 is a chemical probe based on the scaffold of a promiscuous kinase binder, which covalently reacts with a conserved lysine in the ATP-binding pocket of kinases. Pretreatment of cells with a drug (candidate) results in competition of XO44 binding, which can be interpreted as in situ target engagement. Here, a workflow is presented which is optimized for kinome coverage, reproducibility and automated data processing. This dataset pertains to the use of high probe dose samples as peptide library in MaxQuant.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Macrophage

SUBMITTER: Joel Rüegger

LAB HEAD: Mario van

PROVIDER: PXD035540 | Pride | 2025-04-08

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	DMSO_01.raw	Raw
	DMSO_02.raw	Raw
	MV4-11_peptide_library.xlsx	Xlsx
	XO44_10uM_01.raw	Raw
	XO44_10uM_02.raw	Raw

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CellEKT: A chemical proteomics assay to profile cellular target engagement of kinase inhibitors

Project description:Activity-based protein profiling is a powerful tool to study target engagement in complex biological environments. XO44 is a chemical probe based on the scaffold of a promiscuous kinase binder, which covalently reacts with a conserved lysine in the ATP-binding pocket of kinases. Pretreatment of cells with a drug (candidate) results in competition of XO44 binding, which can be interpreted as in situ target engagement. Here, a workflow is presented which is optimized for kinome coverage, reproducibility and data processing. This dataset pertains to the kinome coverage of XO44 in 19 human cell lines as determined by chemical proteomics.

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CellEKT: A chemical proteomics assay to profile cellular target engagement of kinase inhibitors

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CellEKT: A chemical proteomics assay to profile cellular target engagement of kinase inhibitors

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