Unravelling the skin secretion peptides of the gliding leaf frog, Agalychnis spurrelli (hylidae)

Carolina Proaño-Bolaños; Ailín Blasco-Zúñiga; José Rafael Almeida; Lei Wang; Miguel Angel Llumiquinga; Miryan Rivera; Mei Zhou; Tianbao Chen; Chris Shaw

doi:10.3390/biom9110667

Unravelling the skin secretion peptides of the gliding leaf frog, Agalychnis spurrelli (hylidae)

Carolina Proaño-Bolaños^*, Ailín Blasco-Zúñiga, José Rafael Almeida, Lei Wang, Miguel Angel Llumiquinga, Miryan Rivera, Mei Zhou, Tianbao Chen, Chris Shaw

^*Corresponding author for this work

Faculty of Exact and Natural Sciences

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

Frog skin secretions contain medically-valuable molecules, which are useful for the discovery of new biopharmaceuticals. The peptide profile of the skin secretion of Agalychnis spurrelli has not been investigated; therefore, the structural and biological characterization of its compounds signify an inestimable opportunity to acquire new biologically-active chemical scaffolds. In this work, skin secretion from this amphibian was analysed by molecular cloning and tandem mass spectrometry. Although the extent of this work was not exhaustive, eleven skin secretion peptides belonging to five peptide families were identified. Among these, we report the occurrence of two phyllokinins, and one medusin-SP which were previously reported in other related species. In addition, eight novel peptides were identified, including four dermaseptins, DRS-SP2 to DRS-SP5, one phylloseptin-SP1, and three orphan peptides. Phylloseptin-SP1 and dermaseptins-SP2 were identified in HPLC fractions based on their molecular masses determined by MALDI-TOF MS. Among the antimicrobial peptides, dermaseptin-SP2 was the most potent, inhibiting Escherichia coli, Staphylococcus aureus, and ORSA with a minimum inhibitory concentration (MIC) of 2.68 µM, and Candida albicans with an MIC of 10.71 µM, without haemolytic effects. The peptides described in this study represent but a superficial glance at the considerable structural diversity of bioactive peptides produced in the skin secretion of A. spurrelli.

Original language	English
Article number	667
Journal	Biomolecules
Volume	9
Issue number	11
DOIs	https://doi.org/10.3390/biom9110667
State	Published - Nov 2019

Bibliographical note

Publisher Copyright:
© 2019 by the authors. Licensee MDPI, Basel, Switzerland.

Funding

Acknowledgments: Carolina Proaño-Bolaños received a scholarship of the Ecuadorian Secretariat of Science and Technology (SENESCYT). This research was funded by the Natural Drug Discovery Group, School of Pharmacy, Queen’s University Belfast (QUB), the Pontificia Universidad Católica del Ecuador project code QINV0046-IINV529010100, Centro de Investigación para la Salud en América Latina-CISeAL, and SENESCYT. The latter also supported field work to CPB. The collection and rearing of frogs in Ecuador were done under permits of the Ecuadorian Ministerio de Ambiente (MAE): 001-13 IC-FAU-DNB/MA, 003-11 IC-FAU-DNB/MA, and 005-15 IC-FAU-DNB/MA (issued to the Centro Jambatu of Amphibian Research and Conservation, Quito, Ecuador). Exportation of skin secretion samples were done under exportation permits 003-13-EXP-CI-FAU-DNB/MA and 2015-003-FO-DPAP-MA. 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 Sonia Zapata, Iliana Alcocer and Jorge Reyes. Elicio E. Tapia (CJ) aided in collecting specimens in Esmeraldas province, Luis A. Coloma (CJ) reviewed a pre-submitted version of this MS, Berenice Benavides helped with the experiments at CISeAL, Verónica Sánchez was of great help with MIC plots, and Catriona Arlow helped with the language reviews of the first draft of this manuscript. This research was funded by (1) SENESCYT through a scholarship to C.P.B, (2) THE NATURAL DRUG DISCOVERY GROUP of QUB, (3) The PONTIFICIA UNIVERSIDAD CAT?LICA DEL ECUADOR, project code QINV0046-IINV529010100, and (4) The ?CENTRO DE INVESTIGACI?N PARA LA SALUD EN AM?RICA LATINA (CISeAL) which funded APC charges.Carolina Proa?o-Bola?os received a scholarship of the Ecuadorian Secretariat of Science and Technology (SENESCYT). This research was funded by the Natural Drug Discovery Group, School of Pharmacy, Queen?s University Belfast (QUB), the Pontificia Universidad Cat?lica del Ecuador project code QINV0046-IINV529010100, Centro de Investigaci?n para la Salud en Am?rica Latina-CISeAL, and SENESCYT. The latter also supported field work to CPB. The collection and rearing of frogs in Ecuador were done under permits of the Ecuadorian Ministerio de Ambiente (MAE): 001-13 IC-FAU-DNB/MA, 003-11 IC-FAU-DNB/MA, and 005-15 IC-FAU-DNB/MA (issued to the Centro Jambatu of Amphibian Research and Conservation, Quito, Ecuador). Exportation of skin secretion samples were done under exportation permits 003-13-EXP-CI-FAU-DNB/MA and 2015-003-FO-DPAP-MA. 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 Amaz?nica, 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 Sonia Zapata, Iliana Alcocer and Jorge Reyes. Elicio E. Tapia (CJ) aided in collecting specimens in Esmeraldas province, Luis A. Coloma (CJ) reviewed a pre-submitted version of this MS, Berenice Benavides helped with the experiments at CISeAL, Ver?nica S?nchez was of great help with MIC plots, and Catriona Arlow helped with the language reviews of the first draft of this manuscript.

Funders	Funder number
Centro de Investigaci?n para la Salud en Am?rica Latina-CISeAL
Centro de Investigación para la Salud en América Latina-CISeAL
Ecuadorian Ministerio de Ambiente	003-13-EXP-CI-FAU-DNB/MA, 005-15 IC-FAU-DNB/MA, 2015-003-FO-DPAP-MA, 003-11 IC-FAU-DNB/MA, MAE-DNB-CM-2016-0051, 001-13 IC-FAU-DNB/MA
Ecuadorian Secretariat of Science and Technology
Global Environmental Facility
Natural Drug Discovery Group
Pontificia Universidad Católica del Ecuador
Programa de las Naciones Unidas para el Desarrollo?
Queen?s University Belfast
Secretaría de Educación Superior, Ciencia, Tecnología e Innovación

Keywords

Agalychnis spurrelli
Dermaseptins
Frog skin secretion
Peptidomics
Phylloseptins
Tandem mass spectrometry

Access to Document

10.3390/biom9110667

Cite this

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title = "Unravelling the skin secretion peptides of the gliding leaf frog, Agalychnis spurrelli (hylidae)",

abstract = "Frog skin secretions contain medically-valuable molecules, which are useful for the discovery of new biopharmaceuticals. The peptide profile of the skin secretion of Agalychnis spurrelli has not been investigated; therefore, the structural and biological characterization of its compounds signify an inestimable opportunity to acquire new biologically-active chemical scaffolds. In this work, skin secretion from this amphibian was analysed by molecular cloning and tandem mass spectrometry. Although the extent of this work was not exhaustive, eleven skin secretion peptides belonging to five peptide families were identified. Among these, we report the occurrence of two phyllokinins, and one medusin-SP which were previously reported in other related species. In addition, eight novel peptides were identified, including four dermaseptins, DRS-SP2 to DRS-SP5, one phylloseptin-SP1, and three orphan peptides. Phylloseptin-SP1 and dermaseptins-SP2 were identified in HPLC fractions based on their molecular masses determined by MALDI-TOF MS. Among the antimicrobial peptides, dermaseptin-SP2 was the most potent, inhibiting Escherichia coli, Staphylococcus aureus, and ORSA with a minimum inhibitory concentration (MIC) of 2.68 µM, and Candida albicans with an MIC of 10.71 µM, without haemolytic effects. The peptides described in this study represent but a superficial glance at the considerable structural diversity of bioactive peptides produced in the skin secretion of A. spurrelli.",

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author = "Carolina Proa{\~n}o-Bola{\~n}os and Ail{\'i}n Blasco-Z{\'u}{\~n}iga and Almeida, {Jos{\'e} Rafael} and Lei Wang and Llumiquinga, {Miguel Angel} and Miryan Rivera and Mei Zhou and Tianbao Chen and Chris Shaw",

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AU - Proaño-Bolaños, Carolina

AU - Blasco-Zúñiga, Ailín

AU - Almeida, José Rafael

AU - Wang, Lei

AU - Llumiquinga, Miguel Angel

AU - Rivera, Miryan

AU - Zhou, Mei

AU - Chen, Tianbao

AU - Shaw, Chris

PY - 2019/11

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N2 - Frog skin secretions contain medically-valuable molecules, which are useful for the discovery of new biopharmaceuticals. The peptide profile of the skin secretion of Agalychnis spurrelli has not been investigated; therefore, the structural and biological characterization of its compounds signify an inestimable opportunity to acquire new biologically-active chemical scaffolds. In this work, skin secretion from this amphibian was analysed by molecular cloning and tandem mass spectrometry. Although the extent of this work was not exhaustive, eleven skin secretion peptides belonging to five peptide families were identified. Among these, we report the occurrence of two phyllokinins, and one medusin-SP which were previously reported in other related species. In addition, eight novel peptides were identified, including four dermaseptins, DRS-SP2 to DRS-SP5, one phylloseptin-SP1, and three orphan peptides. Phylloseptin-SP1 and dermaseptins-SP2 were identified in HPLC fractions based on their molecular masses determined by MALDI-TOF MS. Among the antimicrobial peptides, dermaseptin-SP2 was the most potent, inhibiting Escherichia coli, Staphylococcus aureus, and ORSA with a minimum inhibitory concentration (MIC) of 2.68 µM, and Candida albicans with an MIC of 10.71 µM, without haemolytic effects. The peptides described in this study represent but a superficial glance at the considerable structural diversity of bioactive peptides produced in the skin secretion of A. spurrelli.

AB - Frog skin secretions contain medically-valuable molecules, which are useful for the discovery of new biopharmaceuticals. The peptide profile of the skin secretion of Agalychnis spurrelli has not been investigated; therefore, the structural and biological characterization of its compounds signify an inestimable opportunity to acquire new biologically-active chemical scaffolds. In this work, skin secretion from this amphibian was analysed by molecular cloning and tandem mass spectrometry. Although the extent of this work was not exhaustive, eleven skin secretion peptides belonging to five peptide families were identified. Among these, we report the occurrence of two phyllokinins, and one medusin-SP which were previously reported in other related species. In addition, eight novel peptides were identified, including four dermaseptins, DRS-SP2 to DRS-SP5, one phylloseptin-SP1, and three orphan peptides. Phylloseptin-SP1 and dermaseptins-SP2 were identified in HPLC fractions based on their molecular masses determined by MALDI-TOF MS. Among the antimicrobial peptides, dermaseptin-SP2 was the most potent, inhibiting Escherichia coli, Staphylococcus aureus, and ORSA with a minimum inhibitory concentration (MIC) of 2.68 µM, and Candida albicans with an MIC of 10.71 µM, without haemolytic effects. The peptides described in this study represent but a superficial glance at the considerable structural diversity of bioactive peptides produced in the skin secretion of A. spurrelli.

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ER -

Unravelling the skin secretion peptides of the gliding leaf frog, Agalychnis spurrelli (hylidae)

Abstract

Bibliographical note

Funding

Keywords

Access to Document

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