Identification, affinity characterisation and biological interactions of lectin-like peptide-carbohydrate complexes derived from human TNF-alpha using high-resolution mass spectrometry

J Pept Sci. 2007 Dec;13(12):803-10. doi: 10.1002/psc.902.


A cyclic disulfide heptadecapeptide (TIP17ox; 2) derived from the lectin-like 17-amino acid domain of human tumor necrosis factor-alpha [TNF-alpha (100-116)] was synthesised and demonstrated to bind specifically to N,N-diacetylchitobiose, a disaccharide present in many glycan structures of glycoproteins. Although the TIP domain forms a loop structure in the native TNF-alpha protein, we show in this study by high-resolution ESI-FTICR mass spectrometry that a homologous linear heptadecapeptide (TIP17rd; 1) binds with comparable affinity to chitobiose, suggesting that cyclisation is not essential for carbohydrate binding. ESI-FTICR-MS was used as an efficient tool for the direct molecular characterisation of TIP peptide-carbohydrate complexes. The specific binding of the TNF-TIP domain to chitobiose and other carbohydrate motifs in glycoproteins may explain the high proteolytic stability of these peptides in biological fluids. A considerably higher proteolytic stability in human plasma was found by mass spectrometric analysis for the cyclic TIP peptide 2, compared to the linear peptide 1. Furthermore, affinity-proteomics studies using immobilised cyclic TIP peptide 2 provided the identification of specific interacting glycoproteins in plasma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Binding, Competitive
  • Carbohydrate Conformation
  • Carbohydrates / chemical synthesis
  • Carbohydrates / chemistry*
  • Carbohydrates / pharmacology
  • Disaccharides / chemistry*
  • Disulfides / chemistry
  • Humans
  • Lectins / chemical synthesis
  • Lectins / chemistry*
  • Lectins / pharmacology
  • Molecular Sequence Data
  • Peptides, Cyclic / chemical synthesis
  • Peptides, Cyclic / chemistry*
  • Peptides, Cyclic / pharmacology
  • Protein Conformation
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Spectroscopy, Fourier Transform Infrared / methods
  • Tumor Necrosis Factor-alpha / chemistry*


  • Carbohydrates
  • Disaccharides
  • Disulfides
  • Lectins
  • Peptides, Cyclic
  • Tumor Necrosis Factor-alpha
  • N,N-diacetylchitobiose