A modular cross-linking approach for exploring protein interactions

J Am Chem Soc. 2003 Mar 5;125(9):2416-25. doi: 10.1021/ja026917a.

Abstract

A method is described for the elucidation of protein-protein interactions using novel cross-linking reagents and mass spectrometry. The method incorporates (1) a modular solid-phase synthetic strategy for generating the cross-linking reagents, (2) enrichment and digestion of cross-linked proteins using microconcentrators, (3) mass spectrometric analysis of cross-linked peptides, and (4) comprehensive computational analysis of the cross-linking data. This integrated approach has been applied to the study of cross-linking between the components of the heterodimeric protein complex negative cofactor 2.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acrylic Resins / chemistry
  • Amino Acid Sequence
  • Biotin / chemistry
  • Cross-Linking Reagents / chemical synthesis
  • Cross-Linking Reagents / chemistry*
  • Glycine / chemistry
  • Models, Molecular
  • Molecular Sequence Data
  • Peptide Fragments / chemistry
  • Phosphoproteins / chemistry*
  • Polyethylene Glycols / chemistry
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
  • Sulfhydryl Compounds / chemistry
  • Transcription Factors / chemistry*

Substances

  • Acrylic Resins
  • Cross-Linking Reagents
  • Peptide Fragments
  • Phosphoproteins
  • Sulfhydryl Compounds
  • Transcription Factors
  • down-regulator of transcription 1
  • Polyethylene Glycols
  • Biotin
  • polyacrylamide
  • Glycine