Chances and pitfalls of chemical cross-linking with amine-reactive N-hydroxysuccinimide esters

Anal Bioanal Chem. 2008 Sep;392(1-2):305-12. doi: 10.1007/s00216-008-2231-5.


In this report we summarize our experiences with the reaction products of N-hydroxysuccinimide (NHS) esters, which are widely used for chemical cross-linking of lysine residues in proteins. We describe the products, which should be scrutinized during data analysis using customized software when NHS esters are employed for chemical cross-linking. Reaction products of NHS esters were observed not only with lysines, but also with serines, tyrosines, and threonines. This report is intended to be a practical guide for those working in the field of chemical cross-linking and mass spectrometry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / chemistry*
  • Cross-Linking Reagents / chemistry*
  • Esters / chemistry
  • Laminin / chemistry
  • Lysine / chemistry
  • Molecular Sequence Data
  • Peptides / analysis
  • Peptides / chemistry*
  • Serine / chemistry
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
  • Succinimides / chemistry*
  • Threonine / chemistry
  • Tyrosine / chemistry


  • Amino Acids
  • Cross-Linking Reagents
  • Esters
  • LAMB1 protein, human
  • Laminin
  • Peptides
  • Succinimides
  • laminin gamma 1
  • Threonine
  • Tyrosine
  • Serine
  • Lysine
  • N-hydroxysuccinimide