SELDI-TOF-MS as a rapid tool to study food related protein–peptide interactions

H.A. Kosters, P.A. Wierenga, H. Gruppen

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Abstract

The use of SELDI-TOF-MS was investigated as a rapid tool to detect peptides present in a crude protein hydrolysate, which are capable to bind to intact food proteins. A purified and well characterized ß-lactoglobulin preparation was extensively hydrolyzed by the Glu-specific enzyme V8 from Staphylococcus aureus. Characterization of this hydrolysate by SELDI-TOF-MS and MALDI-TOF-MS resulted in sixteen identified peptides, which covered 98% of the primary sequence of ß-lactoglobulin. To identify peptides capable to bind non-covalently to intact proteins, the complete hydrolysate was applied to covalently bound ovalbumin, glycinin, ß-lactoglobulin and ß-casein on a SELDI ProteinChip PS-20. Six peptides (AB [f29–45], AB [f90–108], AB [f138–158], B [f63–89], AB [f1–45], AB [f135–162] bound to these four different proteins with decreasing affinity to glycinin > ovalbumin > ß-lactoglobulin > ß-casein. Peptides, which bound to these proteins were AB [f1–45] and AB [f135–158]. Using different concentrations of Triton X-100 (up to 2%) as a washing step prior to MS detection, enabled a rapid distinction between the peptides bound with respect to protein binding capacity.
Original languageEnglish
Pages (from-to)667-673
JournalFood Hydrocolloids
Volume24
Issue number6-7
DOIs
Publication statusPublished - 2010

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Keywords

  • bacillus-licheniformis protease
  • flight-mass spectrometry
  • beta-lactoglobulin
  • aggregation behavior
  • hydrolysis
  • identification
  • fractions
  • isolate
  • stability
  • kinetics

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