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Application of folding stability-based proteomics to profile cancer drug

ABSTRACT: Covalent inhibitors offer powerful therapeutic advantages but require comprehensive profiling to define their on- and off-target activities. Here, we apply two stability-based proteomic methods, SPROX and TPP, to characterize protein targets of the KRASG12C inhibitor ARS-1620 in H358 cell lysate. Both methods identified ligand-induced protein stability shifts, collectively recovering the known on-target KRAS as well as multiple off-targets. Comparative analyses with ABPP and pull-down datasets highlighted ALDH1A3 as a reproducible off-target. Bottom-up proteomics mapped Cys314 as the major ARS-1620 modification site on ALDH1A3, and enzymatic assays confirmed dose-dependent inhibition. Covalent docking supported a favorable binding pose within the retinal-binding pocket. Together, these findings demonstrate that stability-based proteomics provides an effective, derivatization-free strategy for covalent drug target identification.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Non-small Cell Lung Carcinoma

SUBMITTER: You Zou

LAB HEAD: Michael C.

PROVIDER: PXD070862 | Pride | 2026-05-08

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	ALDH1A3.fasta	Fasta
	ALDH1A3_ARS1620.raw	Raw
	ALDH1A3_ARS_1620_psm.tsv	Tabular
	ARS1620_2026_04_26.pdStudy	Other
	ARS_1620_2.raw	Raw

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