Proteomic analysis of soluble proteins retrieved from Duttaphrynus melanostictus skin secretion by IEx-batch sample preparation.

Autor: Mariano DOC; Laboratório de Bioquímica e Biofísica, Instituto Butantan, São Paulo, SP 05503-900, Brazil., Prezotto-Neto JP; Centro de Biotecnologia, IPEN, São Paulo, SP 05508-000, Brazil., Spencer PJ; Centro de Biotecnologia, IPEN, São Paulo, SP 05508-000, Brazil., Sciani JM; Laboratório Multidisciplinar de Pesquisa, Universidade São Francisco, Bragança Paulista, SP 12916-900, Brazil., Pimenta DC; Laboratório de Bioquímica e Biofísica, Instituto Butantan, São Paulo, SP 05503-900, Brazil. Electronic address: dcpimenta@butantan.gov.br.
Jazyk: angličtina
Zdroj: Journal of proteomics [J Proteomics] 2019 Oct 30; Vol. 209, pp. 103525. Date of Electronic Publication: 2019 Sep 14.
DOI: 10.1016/j.jprot.2019.103525
Abstrakt: Amphibians display a toxic secretion that works as chemical defenses against predators and/or microorganisms that is stored in specialized glands located in the tegument. For some animals, such glands have accumulated in specific regions of the body and formed prominent structures known as macroglands. The Bufonidae family displays conspicuous macroglands in a post-orbital position, termed parotoids, which secretions are known to be extremely viscous and rich in alkaloids and steroids. Few proteins have been described in this material, though. Mainly, because of the difficulties to handle such biological matrix. In this context, we have performed a proteomic study on the parotoid macrogland secretion of the Asian bufonid Duttaphrynus melanostictus. By employing the Ion-Exchange (IEx)-batch chromatography (anionic, cationic and both) we obtained six fractions - bound and unbound - that were submitted to an in solution-trypsin digestion followed by LC-MS/MS. Proteins related to: antioxidant activity, binding processes (carbohydrate/lipid/protein), energy metabolism, hydrolases, lipid metabolism and membrane traffic were identified. Moreover, IEx was able to preserve the biological activity of the retrieved proteins (peptidasic). The current study increases the knowledge on the proteins present in the bufonids parotoid macrogland secretion, providing a better understanding of the physiological role played by such molecules. SIGNIFICANCE: The current approach allowed a detailed proteomic analysis of the several proteins synthesized in the D. melanostictus parotoid macrogland (Bufonidae) that are secreted into the skins, but embedded within a complex viscous biological matrix. Moreover, our results aim to increase the knowledge on the biological role played by such proteins at the skin.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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