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Exploring Alternative Cysteine-reactive Chemistry for MS-cleavable Cross-linking Mass Spectrometry

ABSTRACT: To enable global PPI mapping, we have explored an alternative cysteine labeling chemistry, and thus designed and synthesized a sulfoxide-containing MS-cleavable haloacetamide-based cross-linker, DiBromoAcetamide SulfOxide (DBrASO). In combination with a newly developed SEC-HpHt fractionation method, we have successfully applied DBrASO for cross-linking of HEK293 cell lysates and demonstrated its capability to complement lysine-reactive reagents in order to expand PPI coverage at systems-level.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: Fenglong Jiao

LAB HEAD: Lan Huang

PROVIDER: PXD037443 | Pride | 2023-07-20

REPOSITORIES: Pride

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Exploring an Alternative Cysteine-Reactive Chemistry to Enable Proteome-Wide PPI Analysis by Cross-Linking Mass Spectrometry.

Jiao Fenglong F Salituro Leah J LJ Yu Clinton C Gutierrez Craig B CB Rychnovsky Scott D SD Huang Lan L

Analytical chemistry 20230118 4

The development of MS-cleavable cross-linking mass spectrometry (XL-MS) has enabled the effective capture and identification of endogenous protein-protein interactions (PPIs) and their residue contacts at the global scale without cell engineering. So far, only lysine-reactive cross-linkers have been successfully applied for proteome-wide PPI profiling. However, lysine cross-linkers alone cannot uncover the complete PPI map in cells. Previously, we have developed a maleimide-based cysteine-reacti ...[more]

PMID: 36652389

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Integrative Structural Characterization of the Human Canonical and Non-canonical Cop9 Signalosome Complexes

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