Cross-linking mass spectrometry faces challenges due to low reaction efficiencies and complex matrices in protein–protein interaction (PPI) studies. Here, the authors enhance an Azide-A-DSBSO-based in vivo crosslinking workflow with affinity enrichment and size exclusion chromatography, yielding over 5000 crosslinks from K562 cells and generating a comprehensive PPI network.
- Philipp Bräuer
- Laszlo Tirian
- Manuel Matzinger