Molecular cloning of the trypsin inhibitor from the skin secretion of the Madagascan Tomato Frog, Dyscophus guineti (Microhylidae), and insights into its potential defensive role

König, Enrico; Wesse, Christina; Murphy, Anna; Wang, Lei; Shaw, Chris; Bininda‐Emonds, Olaf R. P.

doi:10.1007/s13127-013-0128-4

Cited by 4 publications

(5 citation statements)

References 34 publications

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“…Besides providing an overview of expressed genes, this library served as a database to analyse proteomic (mass spectrometry; MS) data obtained from glue samples (produced by dorsal skin) and compare them to those of mucus samples (produced by limb skin). Using this approach, we found transcriptional confirmation of the previously described D. guineti serine protease inhibitor 21 , 22 . However, proteomic analyses did not substantiate the presence of the serine protease inhibitor in either the glue or mucus, perhaps because of our choice of proteomic methods (see Methods) or due to the previously reported low levels of this protein in the secreted material 22 .…”

Section: Resultssupporting

confidence: 63%

“…Using this approach, we found transcriptional confirmation of the previously described D. guineti serine protease inhibitor 21 , 22 . However, proteomic analyses did not substantiate the presence of the serine protease inhibitor in either the glue or mucus, perhaps because of our choice of proteomic methods (see Methods) or due to the previously reported low levels of this protein in the secreted material 22 . Similarly, we found no evidence for the secretion of any bioactive peptides shared by a wide range of other frog families (including neuropeptides, hormone analogs or cytolytic peptides 11 ).…”

Section: Resultssupporting

confidence: 63%

“…A proteinaceous nature of the glues of the frog species Notaden bennetti (Myobatrachidae) 13 and Eupsophus vertebralis (Alsodidae) 15 has been demonstrated and while a protein called Nb1 has been reported for the former species, it was never characterised in detail. Studies on the adhesive secretion of another glue-secreting amphibian, the Madagascan tomato frog Dyscophus guineti (Microhylidae), identified one of its constituents as a serine protease inhibitor 21 , 22 . Its role in the glue, however, remains speculative 22 .…”

Section: Introductionmentioning

confidence: 99%

“…Studies on the adhesive secretion of another glue-secreting amphibian, the Madagascan tomato frog Dyscophus guineti (Microhylidae), identified one of its constituents as a serine protease inhibitor 21 , 22 . Its role in the glue, however, remains speculative 22 .…”

Section: Introductionmentioning

confidence: 99%

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Recurrent evolution of adhesive defence systems in amphibians by parallel shifts in gene expression

Zaman,

Lengerer,

Van Lindt

et al. 2024

Nat Commun

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Natural selection can drive organisms to strikingly similar adaptive solutions, but the underlying molecular mechanisms often remain unknown. Several amphibians have independently evolved highly adhesive skin secretions (glues) that support a highly effective antipredator defence mechanism. Here we demonstrate that the glue of the Madagascan tomato frog, Dyscophus guineti, relies on two interacting proteins: a highly derived member of a widespread glycoprotein family and a galectin. Identification of homologous proteins in other amphibians reveals that these proteins attained a function in skin long before glues evolved. Yet, major elevations in their expression, besides structural changes in the glycoprotein (increasing its structural disorder and glycosylation), caused the independent rise of glues in at least two frog lineages. Besides providing a model for the chemical functioning of animal adhesive secretions, our findings highlight how recruiting ancient molecular templates may facilitate the recurrent evolution of functional innovations.

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Section: Resultssupporting

confidence: 63%

Section: Resultssupporting

confidence: 63%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Recurrent evolution of adhesive defence systems in amphibians by parallel shifts in gene expression

Zaman,

Lengerer,

Van Lindt

et al. 2024

Nat Commun

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“…Due to vast amphibian biodiversity, diverse applications and an enormous array of bioactive molecules that have not been described yet, frog and toad (Anurans) skin secretions continue drawing attention as source of novel biologically active molecules [ 4 , 8 ]. Although peptides of 10 to 48 residues long comprise the majority of biomolecules described in literature [ 8 ], a considerable number of species from distinct families have been described to lack peptides in their skin secretions [ 9 , 10 , 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

Identification of Arenin, a Novel Kunitz-Like Polypeptide from the Skin Secretions of Dryophytes arenicolor

Hernández-Pérez

Serra

Sze

et al. 2018

IJMS

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Amphibian skin secretions are enriched with complex cocktails of bioactive molecules such as proteins, peptides, biogenic amines, alkaloids guanidine derivatives, steroids and other minor components spanning a wide spectrum of pharmacological actions exploited for centuries in folk medicine. This study presents evidence on the protein profile of the skin secretions of the canyon tree frog, Dryophytes arenicolor. At the same time, it presents the reverse-phase liquid chromatography isolation, mass spectrometry characterization and identification at mRNA level of a novel 58 amino acids Kunitz-like polypeptide from the skin secretions of Dryophytes arenicolor, arenin. Cell viability assays performed on HDFa, CaCo2 and MCF7 cells cultured with different concentrations of arenin showed a discrete effect at low concentrations (2, 4, 8 and 16 µg/mL) suggesting a multi-target interaction in a hormetic-like dose-response. Further work is required to investigate the mechanisms underlying the variable effect on cell viability produced by different concentrations of arenin.

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Biochemical Analyses of Proteins from Duttaphrynus melanostictus (Bufo melanostictus) Skin Secretion: Soluble Protein Retrieval from a Viscous Matrix by Ion-Exchange Batch Sample Preparation

et al. 2018

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A crucial step in scientific analysis can be sample preparation, and its importance increases in the same rate as the sensitivity of the following employed/desired analytical technique does. The need to analyze complex, viscous matrices is not new, and diverse approaches have been employed, with different success rates depending on the intended molecules. Solid-phase extraction, for example, has been successfully used in sample preparation for organic molecules and peptides. However, due to the usual methodological conditions, biologically active proteins are not successfully retrieved by this technique, resulting in a low rate of protein identification reported for the viscous amphibian skin secretion. Here we describe an ion-exchange batch processing sample preparation technique that allows viscous or adhesive materials (as some amphibian skin secretions) to be further processed by classical liquid chromatography approaches. According to our protocol, samples were allowed to equilibrate with a specific resin that was washed with appropriated buffers in order to yield the soluble protein fraction. In order to show the efficiency of our methodology, we have compared our results to classically prepared skin secretion, i.e., by means of filtration and centrifugation. After batch sample preparation, we were able to obtain reproductive resolved protein chromatographic profiles, as revealed by SDS-PAGE, and retrieve some biological activities, namely, hydrolases belonging to serine peptidase family. Not only that, but also the unbound fraction was rich in low molecular mass molecules, such as alkaloids and steroids, making this sample preparation technique also suitable for the enrichment of such molecules.

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Molecular cloning of the trypsin inhibitor from the skin secretion of the Madagascan Tomato Frog, Dyscophus guineti (Microhylidae), and insights into its potential defensive role

Cited by 4 publications

References 34 publications

Recurrent evolution of adhesive defence systems in amphibians by parallel shifts in gene expression

Recurrent evolution of adhesive defence systems in amphibians by parallel shifts in gene expression

Identification of Arenin, a Novel Kunitz-Like Polypeptide from the Skin Secretions of Dryophytes arenicolor

Biochemical Analyses of Proteins from Duttaphrynus melanostictus (Bufo melanostictus) Skin Secretion: Soluble Protein Retrieval from a Viscous Matrix by Ion-Exchange Batch Sample Preparation

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