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In Situ Cross-linking Based Co-Fractionation Mass Spectrometry (XL-CoFrac-MS)

ABSTRACT: We developed an in situ cross-linking coupled reversed-phase liquid chromatography-based co-fractionation mass spectrometry (XL-CoFrac-MS) platform to analyze drug-induced protein complex changes. A software tool, ChromaQuant, was built to automate chromatographic peak quantification. Using RS4;11 leukemia cells treated with the BCL-2 inhibitor ABT-199, we demonstrated the ability of the platform to detect BCL-2-associated protein complexes and characterize signaling pathway alterations. This approach enables efficient protein complex profiling and provides mechanistic insights into drug-induced apoptosis.

ORGANISM(S): Homo Sapiens

SUBMITTER: Mingxia Gao

PROVIDER: PXD067673 | iProX | Sat Aug 23 00:00:00 GMT+01:00 2025

REPOSITORIES: iProX

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A Chromatography-Guided Co-Fractionation Mass Spectrometry Strategy for Rapid Profiling of Drug-Perturbed Protein Complexes.

Liu Wei W Tang Jiayu J Yan Guoquan G Shen Shun S Gao Mingxia M Zhang Xiangmin X

Analytical chemistry 20250829 35

Protein complexes are central to cellular function and respond rapidly to pharmacological perturbations. Co-fractionation mass spectrometry (CoFrac-MS) is widely employed to analyze protein complexes by analyzing individual chromatographic fractions, but it is labor-intensive and slow. To address these challenges, we introduce a chromatography-guided strategy enabling rapid identification of drug-perturbed protein complexes. It combines cross-linking enhanced reversed phase liquid chromatography ...[more]

PMID: 40878671

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