Application of mass spectrometry (MS) in the structural analysis of selected polypeptides of boar seminal plasma obtained after two-dimensional electrophoresis (2-D PAGE)*

Two-dimensional electrophoresis (2-D PAGE) led to identification in the polypeptide maps of boar seminal plasma of four conserved polypeptides with identical molecular weight of 24 kDA, and different ranges of isoelectric point (pI): (1) 7.4-7.7, (2) 8.1-8.4, (3) 8.5-8.8 and (4) 9.2-9.4. In the current study the molecular structures of these polypeptides were analysed, for the first time, by mass spectrometry (LC – MS/MS). Computerized mass spectrometry analysis of the peptides obtained after trypsin-digestion of the polypeptides demonstrated their similarity to the family of spermadhesins (crystal structure of two members of the spermadhesin family), especially to epididymal spermadhesin AWN-1. In addition, homology was found of peptides 3 and 4 with a lysozyme C precursor (1,4-beta-N-acetylmuramidase C). The results presented might indicate the participation of the analysed polypeptides in the processes accompanying fertilization.