Peptidomic analysis of the skin secretions of the pickerel frog Rana palustris identifies six novel families of structurally-related peptides

Abstract

Peptidomics methodology has been used to identify and characterize structurally 26 previously undescribed peptides in the electrically stimulated skin secretions of the North American pickerel frog Rana palustris. Peptides in the secretions were analyzed by electrospray mass spectrometry and components in mass range 700–2500 Da, present in major abundance, were purified by reverse-phase HPLC. Cysteine-containing components were identified by treatment with dithiothreitol and 4-vinylpyridine and re-analysis of the derivatizated peptide by mass spectrometry. Application of these techniques led to the identification of six families of structurally-related peptides comprising (a) six peptides containing the consensus sequence Cys-Trp-Xaa-Thr-Lys-Ser-Ile-Pro-Pro-Lys/Arg-Xaa-Cys, (b) three peptides containing the consensus sequence Pro-Pro-Gly-Val-Cys-(Xaa) 3-Lys/Arg-Arg-Cys, (c) two peptides containing the consensus sequence Ser-Phe-His-Val-Phe-Pro-Pro-Trp-Met-Cys-Lys-Xaa-Leu-Lys-Lys-Cys, (d) two peptides containing the consensus sequence Arg-Xaa-Cys-Trp-Lys-(Xaa) 2-Asn-(Xaa) 3-Val-Cys-Ser, (e) nine peptides containing the consensus sequence Ser-Leu-Pro-Ala-Gly-Leu-Ser-Pro, and (f) four peptides containing the consensus sequence Asp-Xaa-Gln-Asp-Arg-Trp-Xaa-Pro. The peptides did not inhibit the growth of Escherichia coli or Staphylococcus aureus and were inactive on hamster vascular or gastric smooth muscle preparations so that their biological functions, if any, remain to be established.