Analysis of protein structure by cross-linking combined with mass spectrometry

Methods Mol Biol. 2014:1156:447-63. doi: 10.1007/978-1-4939-0685-7_30.


Cross-linking combined with mass spectrometry is a powerful technique to study protein structure. Here, we present an optimized protocol for the preparation, processing, and analysis of a protein sample cross-linked with isotopically coded, affinity-enrichable, and CID-cleavable cross-linker CyanurBiotinDimercaptoPropionylSuccinimide using LC/ESI-MS/MS on a Thermo Scientific Orbitrap mass spectrometer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid
  • Cross-Linking Reagents / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Proteins / chemistry*
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Tandem Mass Spectrometry / methods*


  • Cross-Linking Reagents
  • Proteins