Purification, conformational analysis and cytotoxic activities of host-defense peptides from the Tungara frog Engystomops pustulosus (Leptodactylidae; Leiuperinae)

J Michael Conlon, Laure Guilhaudis, Samir Attoub, Laurent Coquet, Jérôme Leprince, Thierry Jouenne, Milena Mechkarska

Research output: Contribution to journalArticlepeer-review


The amphibian family Leptodactylidae is divided into three sub-families: Leiuperinae, Leptodactylinae, and Paratelmatobiinae. Host-defense peptides (HDPs) present in the skins of frogs belonging to the Leptodactylinae have been studied extensively, but information is limited regarding peptides from Leiuperinae species. Peptidomic analysis of norepinephrine-stimulated skin secretions from the Tungara frog Engystomops pustulosus (Leiuperinae) collected in Trinidad led to the isolation and structural characterization of previously undescribed pustulosin-1 (FWKADVKEIG KKLAAKLAEELAKKLGEQ), [Q28E] pustulosin-1 (pustulosin-2), and pustulosin-3 (DWKETAKELLKKIGAKVAQVISDKLNPAPQ). The primary structures of these peptides do not resemble those of previously described frog skin HDPs. In addition, the secretions contained tigerinin-1EP (GCKTYLIEPPVCT) with structural similarity to the tigerinins previously identified in skin secretions from frogs from the family Dicroglossidae. Pustulosin-1 and -3 adopted extended α-helical conformations in 25% trifluoroethanol-water and in the presence of cell membrane models (sodium dodecylsulfate and dodecylphosphocholine micelles). Pustulosin-1 and -3 displayed cytotoxic activity against a range of human tumor-derived cell lines (A549, MDA-MB-231, and HT29), but their therapeutic potential for development into anti-cancer agents is limited by their comparable cytotoxic activity against non-neoplastic human umbilical vein endothelial cells. The peptides also displayed weak antimicrobial activity against Escherichia coli (MIC = 125 µM) but were inactive against Staphylococcus aureus. Tigerinin-1EP was inactive against both the tumor-derived cells and bacteria. [Abstract copyright: © 2023. The Author(s).]
Original languageEnglish
Pages (from-to)1349-1359
Number of pages11
JournalAmino Acids
Issue number10
Early online date7 Aug 2023
Publication statusPublished (in print/issue) - Oct 2023

Bibliographical note

Funding Information:
This work was partially supported by the Université de Rouen Normandie (URN), INSA Rouen Normandie, the Centre National de la Recherche Scientifique (CNRS), European Regional Development Fund (ERDF), Labex SynOrg (ANR-11-LABX-0029), Carnot Institute (I2C), and the graduate School for research XL-Chem (ANR-18-EURE-0020 XL CHEM), and by the UWI Campus Research and Publication Fund (CRP.3.NOV16.8(1)). MM was awarded a Study and Travel grant and Institutional Visit Allowance for a visit to Ulster University (N. Ireland).

Publisher Copyright:
© 2023, The Author(s).


  • Host-defense peptide
  • Frog skin
  • Pustulosin
  • Tigerinin
  • Cytotoxic


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