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

In-Depth In Vivo Crosslinking in Minutes by a Compact, Membrane-Permeable, and Alkynyl-Enrichable Crosslinker.

Gao H, Zhao L, Zhong B, Zhang B, Gong Z, Zhao B, Liu Y, Zhao Q, Zhang L, Zhang Y

Chemical crosslinking coupled with mass spectrometry (CXMS) has emerged as a powerful technique to obtain the dynamic conformations and interaction interfaces of protein complexes. Limited by the poor cell membrane permeability, chemical reactivity, and biocompatibility of crosslinkers, in vivo crosslinking to capture the dynamics of protein complexes with finer temporal resolution and higher coverage is attractive but challenging. In this ... [more]

Anal Chem May. 31, 2022; 94(21);7551-7558 [Pubmed: 35575683]

Throughput

  • High Throughput

Additional Notes

  • High confidence crosslinked peptides were identified with the FDR set to 0.01 at the peptide-spectrum matches (PSM) level and PSMs >= 2.
  • MS non-cleavable crosslinker of bis(succinimidyl) with propargyl tag (BSP)

Related interactions

InteractionExperimental Evidence CodeDatasetThroughputScoreCurated ByNotes
KRT18 DSP
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
3359262
KRT18 DSP
Proximity Label-MS
Proximity Label-MS

An interaction is inferred when a bait-enzyme fusion protein selectively modifies a vicinal protein with a diffusible reactive product, followed by affinity capture of the modified protein and identification by mass spectrometric methods.

High13.4BioGRID
2991857

Curated By

  • BioGRID