Chemical cross-linking and mass spectrometry for mapping three-dimensional structures of proteins and protein complexes

J Mass Spectrom. 2003 Dec;38(12):1225-37. doi: 10.1002/jms.559.


Chemical cross-linking of proteins, an established method in protein chemistry, has gained renewed interest in combination with mass spectrometric analysis of the reaction products for elucidating low-resolution three-dimensional protein structures and interacting sequences in protein complexes. The identification of the large number of cross-linking sites from the complex mixtures generated by chemical cross-linking, however, remains a challenging task. This review describes the most popular cross-linking reagents for protein structure analysis and gives an overview of the strategies employing intra- or intermolecular chemical cross-linking and mass spectrometry. The various approaches described in the literature to facilitate detection of cross-linking products and also computer software for data analysis are reviewed. Cross-linking techniques combined with mass spectrometry and bioinformatic methods have the potential to provide the basis for an efficient structural characterization of proteins and protein complexes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Cross-Linking Reagents / chemistry*
  • Isotope Labeling
  • Macromolecular Substances
  • Mass Spectrometry / methods*
  • Molecular Structure
  • Peptide Mapping
  • Protein Conformation
  • Proteins / chemistry*
  • Proteins / metabolism
  • Software


  • Cross-Linking Reagents
  • Macromolecular Substances
  • Proteins