An integrated workflow for cross-linking/mass spectrometry

We present a concise workflow to enhance the mass spectrometric detection of cross-linked peptides by introducing sequential digestion and the cross-link identification software Xi. Sequential digestion enhances peptide detection by shortening long tryptic peptides while avoiding over-digestion. We demonstrate our simple 12-fraction protocol for cross-linked multi-protein complexes and cell lysates, quantitative analysis, and high-density cross-linking, without requiring specific cross-linker features. This overall approach reveals dynamic protein-protein interaction sites, which are accessible, have fundamental functional relevance and are therefore ideally suited for the development of small molecule inhibitors.

Download data

• Downloaded 2,757 times
• Download rankings, all-time:
  • Site-wide: 4,864
  • In biochemistry: 80
• Year to date:
  • Site-wide: 24,121
• Since beginning of last month:
  • Site-wide: 29,447

Altmetric data

Downloads over time

Distribution of downloads per paper, site-wide