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

Trioxane-based MS-cleavable cross-linking mass spectrometry for profiling multimeric interactions of cellular networks.

Yu C, Novitsky E, Balasubramani SG, Wang X, Shen X, Yang Q, Rychnovsky S, Echeverria I, Huang L

Cross-linking mass spectrometry (XL-MS) is a powerful technology for mapping protein-protein interactions (PPIs) at the systems level. While bivalent cross-links are effective for defining protein interactions and structures, multivalent cross-links offer enhanced spatial resolution to facilitate characterization of heterogeneous protein complexes. However, their identification remains challenging due to fragmentation complexity and the vast expansion of database search space. Here, we present ... [more]

Nat Commun Jul. 01, 2025; 16(1);5585 [Pubmed: 40593561]

Throughput

  • High Throughput

Ontology Terms

  • hek-293 cell (BTO:0000007) [kidney cell line (BTO:0000067)]

Additional Notes

  • In vivo TSTO cross-linking of HEK293 cells
  • Supplementary Data 3

Related interactions

InteractionExperimental Evidence CodeDatasetThroughputScoreCurated ByNotes
TUBA1B TUBB4B
Affinity Capture-MS
Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

High-BioGRID
3374210
TUBB4B TUBA1B
Affinity Capture-MS
Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

High-BioGRID
3374311
TUBA1B TUBB4B
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.

High0.974BioGRID
744867
TUBA1B TUBB4B
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
3447844

Curated By

  • BioGRID