Abstract
Antimicrobial peptides (AMPs) constitute part of a broad range of bioactive compounds present on diverse organisms, including frogs. Peptides, produced in the granular glands of amphibian skin, constitute a component of their innate immune response, providing protection against pathogenic microorganisms. In this work, two novel cruzioseptins peptides, cruzioseptin-16 and -17, extracted from the splendid leaf frog Cruziohyla calcarifer are presented. These peptides were identified using molecular cloning and tandem mass spectrometry. Later, peptides were synthetized using solid-phase peptide synthesis, and their minimal inhibitory concentration and haemolytic activity were tested. Furthermore, these two cruzioseptins plus three previously reported (CZS-1, CZS-2, CZS-3) were computationally characterized. Results show that cruzioseptins are 21–23 residues long alpha helical cationic peptides, with antimicrobial activity against E. coli, S. aureus, and C. albicans and low haemolytic effect. Docking results agree with the principal action mechanism of cationic AMPs that goes through cell membrane disruption due to electrostatic interactions between cationic residues in the cruzioseptins and negative phosphate groups in the pathogen cell membrane. An action mechanism through enzymes inhibition was also tried, but no conclusive results about this mechanism were obtained.
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Acknowledgements
This research was done under the framework contract for genetic resources access between MAE and IKIAM (MAE-DNB-CM-2016-0051) and is part of the project “Conservation of Ecuadorian amphibian diversity and sustainable use of its genetic resources,” which involves MAE, IKIAM-Universidad Regional Amazónica, Queen’s University Belfast, and Centro Jambatu, and the help of the Global Environmental Facility (GEF) and “Programa de las Naciones Unidas para el Desarrollo” (PNUD). We thank the kindly donation of bacterial and fungal strains by Dr. Jorge Reyes (INSPI).
Funding
This research was funded by research grants to L.M. and MR of the Dirección General Académica of the Pontificia Universidad Católica del Ecuador, projects: QINV0180-IINV529020200 and QINV0209-IINV529010100. This research was partially funded by the GEF Project ID 5534 “Conservation of Ecuadorian Amphibian Diversity and Sustainable Use of its Genetic Resources”, Ministry of Environment of Ecuador and Universidad Regional Amazonica IKIAM.
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Cuesta, S.A., Reinoso, C., Morales, F. et al. Novel antimicrobial cruzioseptin peptides extracted from the splendid leaf frog, Cruziohyla calcarifer. Amino Acids 53, 853–868 (2021). https://doi.org/10.1007/s00726-021-02986-w
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DOI: https://doi.org/10.1007/s00726-021-02986-w