A Comprehensive Proteomic Study of the Skin Secretions of the Frog Lithobates spectabilis

Title:A Comprehensive Proteomic Study of the Skin Secretions of the Frog Lithobates spectabilis

Volume: 23 Issue: 7

Author(s): Griselda Demesa-Balderrama, Erika P. Meneses, Lorena Hernández-Orihuela, Victoria Pando- Robles, Maria C. Rodriguez, Carolina Barrientos-Salcedo, Manuel B. Aguilar and Cesar V. F. Batista

Affiliation:

Keywords: Lithobates spectabilis, proteomics, mass spectrometry, antimicrobial, epidermal homeostasis.

Abstract: Disulfide C-terminal loop fragments derived from AMPs and the presence of peptidases have been previously reported in the skin secretions of different amphibians. However, there are only a few studies on the identification of enzymes in frog skin secretion based on the primary structure of these proteins. Similarly, little data exist regarding the identification of disulfide C-terminal loops at large scale. Therefore, a comprehensive study on this issue certainly could bring in much more information for understanding this molecular process and its biochemical consequences. Thus, the aim of this work was to characterize the presence of disulfide C-terminal loop fragments of AMPs and identify the proteins and probable enzymes present in the completely unknown secretion contents of the frog Lithobates spectabilis. For this purpose, high-resolution mass spectrometry was applied to analyze the skin secretions processed by two different protocols: (1) using a cocktail of enzymatic inhibitors and 2) without any protease inhibitors, maintaining the solution for 2 hours at 10°C. Results from procedure-1, revealed 122 molecular masses, whereas procedure-2 permitted 253 different molecular masses to be identified. Fifty-nine peptides including 22 disulfide C-terminal loop-containing peptides were obtained following procedure-2. Polyacrylamide gel electrophoresis separation, tryptic digestion and LCMS/ MS were used for “de novo” sequencing of 111 different peptides and the unequivocal identification of fifteen proteins including at least three different peptidases. Additionally, it was possible to fully sequence eight peptides, including a ranatuerin-related peptide identified here as Spectabilin, that was subsequently chemically synthesized and showed high antibacterial, antiparasitic and cytotoxic activities.

Cite this article as:

Demesa-Balderrama Griselda, P. Meneses Erika, Hernández-Orihuela Lorena, Pando- Robles Victoria, C. Rodriguez Maria, Barrientos-Salcedo Carolina, B. Aguilar Manuel and V. F. Batista Cesar, A Comprehensive Proteomic Study of the Skin Secretions of the Frog Lithobates spectabilis, Protein & Peptide Letters 2016; 23 (7) . https://dx.doi.org/10.2174/0929866523666160505120853

DOI https://dx.doi.org/10.2174/0929866523666160505120853	Print ISSN 0929-8665
Publisher Name Bentham Science Publisher	Online ISSN 1875-5305