Investigation of skin secretory peptidome of Rana lessonae frog by mass spectrometry

Amphibian skin secretion represents a cerain scientific interest as a source of biologically active natural peptides. In the present research skin peptidome of wide-spread European frog Rana lessonae (Camerano, 1882) was studied for the first time ever. Peptide sequencing was accomplished with Fourier transform ion cyclotron resonance mass spectrometer in collision-induced and electron capture dissociation modes. A portion of amphibian peptides contains intramolecular C-terminal disulfide cycle which obstructs mass spectrometric sequencing. Two methods were utilized to overcome this difficulty: reduction with dithiotreithol followed by thiol group alkylation and oxidation into sulfonic acid groups with performic acid. Integrated approach employed in the present study allowed the identification of 49 peptides (of 6 to 37 amino acid residues), including 19 novel species.