Protein identification from the parotoid macrogland secretion of Duttaphrynus melanostictus

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08/20/2019

Douglas Oscar Ceolin Mariano, Marcela Di Giacomo Messias, Patrick Jack Spencer, Daniel Carvalho Pimenta

Journal of Venomous Animals and Toxins including Tropical Diseases 2019;25:2019-0029
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992019000100318&lng=en&nrm=iso&tlng=en | © The Author(s). 2019
Received: May 06, 2019 | Accepted: July 11, 2019 | Published: August 19, 2019

ABSTRACT

Background

Bufonid parotoid macrogland secretion contains several low molecular mass molecules, such as alkaloids and steroids. Nevertheless, its protein content is poorly understood. Herein, we applied a sample preparation methodology that allows the analysis of viscous matrices in order to examine its proteins.

Methods

Duttaphrynus melanostictus parotoid macrogland secretion was submitted to ion-exchange batch sample preparation, yielding two fractions: salt-displaced fraction and acid-displaced fraction. Each sample was then fractionated by anionic-exchange chromatography, followed by in-solution proteomic analysis.

Results

Forty-two proteins could be identified, such as acyl-CoA-binding protein, alcohol dehydrogenase, calmodulin, galectin and histone. Moreover, de novo analyses yielded 153 peptides, whereas BLAST analyses corroborated some of the proteomic-identified proteins. Furthermore, the de novo peptide analyses indicate the presence of proteins related to apoptosis, cellular structure, catalysis and transport processes.

Conclusions

Proper sample preparation allowed the proteomic and de novo identification of different proteins in the D. melanostictus parotoid macrogland secretion. These results may increase the knowledge about the universe of molecules that compose amphibian skin secretion, as well as to understand their biological/physiological role in the granular gland.

PubMed Central https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6707386/

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KEYWORDS

Amphibian skin secretion
Bufonidae
Duttaphrynus melanostictus
Proteomics
Batch chromatography
Asian common toad
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