Cross-Linking-MS (XL-MS)

An interaction is detected between two proteins using chemically reactive or photo-activatable cross-linking reagents that covalently link amino acids in close proximity, followed by mass spectrometry analysis to identify the linked peptides (reviewed in PMID 37406423, 37104977). Experiments may be carried with live cells or cell lysates in which all proteins are expressed at endogenous levels (e.g. PMID 34349018, 35235311) or with recombinant proteins (e.g., PMID 28537071).

Publication

Cross-Linking/Mass Spectrometry Combined with Ion Mobility on a timsTOF Pro Instrument for Structural Proteomics.

Ihling CH, Piersimoni L, Kipping M, Sinz A

The combination of cross-linking/mass spectrometry (XL-MS) and ion mobility is still underexplored for conducting protein conformational and protein-protein interaction studies. We present a method for analyzing cross-linking mixtures on a timsTOF Pro mass spectrometer that allows separating ions based on their gas-phase mobilities. Cross-linking was performed with three urea-based MS-cleavable cross-linkers that deliver distinct fragmentation patterns for cross-linked species upon ... [more]

Anal Chem Aug. 24, 2021; 93(33);11442-11450 [Pubmed: 34375526]

Throughput

  • High Throughput

Ontology Terms

  • hek-293 cell (BTO:0000007) [embryonic kidney cell line (BTO:0002733)]
  • kidney epithelial cell (CL:0002518)
  • kidney (BTO:0000671)

Additional Notes

  • HEK-293 nuclear extract

Related interactions

InteractionExperimental Evidence CodeDatasetThroughputScoreCurated ByNotes
NCL HNRNPA2B1
Co-fractionation
Co-fractionation

Interaction inferred from the presence of two or more protein subunits in a partially purified protein preparation. If co-fractionation is demonstrated between 3 or more proteins, then add them as a complex.

High-BioGRID
3439399

Curated By

  • BioGRID