Deep sequencing analysis of toad Rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretion

Autor(es)

Tanaka Shibao, Priscila Yumi ^[1] ; Cologna, Camila Takeno ^[1] ; Morandi-Filho, Romualdo ^[2] ; Wiezel, Gisele Adriano ^[1] ; Fujimura, Patricia Tiemi ^[2] ; Ueira-Vieira, Carlos ^[2] ; Arantes, Eliane Candiani ^{[3, 2]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Lab Anim Toxins, Sch Pharmaceut Sci Ribeirao Preto, Ave Cafe S-N, Ribeirao Preto, SP - Brazil

^[2] Univ Fed Uberlandia, Lab Genet LABGEN, Inst Genet & Biochem, Campus Umuarama, Ave Para, BR-1720 Uberlandia, MG - Brazil

^[3] Univ Sao Paulo, Dept Phys & Chem, Sch Pharmaceut Sci Ribeirao Preto, Av Cafe S-N, BR-14040903 Ribeirao Preto, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of Venomous Animals and Toxins including Tropical Diseases; v. 24, NOV 29 2018.
• Citações Web of Science: 1

Resumo

BackgroundAnimal poisons and venoms are sources of biomolecules naturally selected. Rhinella schneideri toads are widespread in the whole Brazilian territory and they have poison glands and mucous gland. Recently, protein from toads' secretion has gaining attention. Frog skin is widely known to present great number of host defense peptides and we hypothesize toads present them as well. In this study, we used a RNA-seq analysis from R. schneideri skin and biochemical tests with the gland secretion to unravel its protein molecules.MethodsTotal RNA from the toad skin was extracted using TRizol reagent, sequenced in duplicate using Illumina Hiseq2500 in paired end analysis. The raw reads were trimmed and de novo assembled using Trinity. The resulting sequences were submitted to functional annotation against non-redundant NCBI database and Database of Anuran Defense Peptide. Furthermore, we performed caseinolytic activity test to assess the presence of serine and metalloproteases in skin secretion and it was fractionated by fast liquid protein chromatography using a reverse-phase column. The fractions were partially sequenced by Edman's degradation.ResultsWe were able to identify several classes of antimicrobial peptides, such as buforins, peroniins and brevinins, as well as PLA(2), lectins and galectins, combining protein sequencing and RNA-seq analysis for the first time. In addition, we could isolate a PLA(2) from the skin secretion and infer the presence of serine proteases in cutaneous secretion.ConclusionsWe identified novel toxins and proteins from R. schneideri mucous glands. Besides, this is a pioneer study that presented the in depth characterization of protein molecules richness from this toad secretion. The results obtained herein showed evidence of novel AMP and enzymes that need to be further explored. (AU)

• Processo FAPESP: 13/26200-6 - Plataforma venômica: proteoma, transcriptoma, clonagem e expressão de toxinas de interesse biotecnológico presentes na peçonha de Pachycondyla villosa
• Beneficiário: Camila Takeno Cologna
• Linha de fomento: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 15/18432-0 - Bioprospecção de toxinas animais com interesse biotecnológico a partir de ferramentas ômicas
• Beneficiário: Eliane Candiani Arantes Braga
• Linha de fomento: Auxílio à Pesquisa - Regular
• Processo FAPESP: 14/15644-3 - Transcriptoma da glândula mucosa de Rhinella schneideri
• Beneficiário: Priscila Yumi Tanaka Shibao
• Linha de fomento: Bolsas no Brasil - Mestrado