Characterisation of phosvitin phosphopeptides using MALDI-TOF mass spectrometry.

Putative phosphopeptides produced from enzyme hydrolysis of phosvitin were identified and characterised using MALDI-TOF/MS. Phosvitin was heat-pretreated and then hydrolysed using pepsin, thermolysin, and trypsin at their optimal pH and temperature conditions with or without partial dephosphorylation. Pepsin and thermolysin were not effective in producing phosphopeptides, but trypsin hydrolysis produced many peptides from phosvitin: 12 peptides, 10 of which were phosphopeptides, were identified from the trypsin hydrolysate. Twelve peptides were also identified from the trypsin hydrolysate of partially dephosphorylated phosvitin, but the phosphate groups remaining with the peptides were much smaller than those from the trypsin hydrolysate of intact phosvitin. This suggested that the phosphopeptides produced from the partially dephosphorylated phosvitin lost most of their phosphate groups during the dephosphorylation step. Therefore, partial dephosphorylation of phosvitin before trypsin hydrolysis may not be always recommendable in producing functional phosphopeptides if the phosphate groups play important roles for their functionalities.