Deep sequencing analysis of toad Rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretion
Abstract Background Animal 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 kno...
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| Format: | Article |
| Language: | English |
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SciELO
2018-11-01
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| Series: | Journal of Venomous Animals and Toxins including Tropical Diseases |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s40409-018-0173-8 |
| _version_ | 1818321916119744512 |
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| author | Priscila Yumi Tanaka Shibao Camila Takeno Cologna Romualdo Morandi-Filho Gisele Adriano Wiezel Patricia Tiemi Fujimura Carlos Ueira-Vieira Eliane Candiani Arantes |
| author_facet | Priscila Yumi Tanaka Shibao Camila Takeno Cologna Romualdo Morandi-Filho Gisele Adriano Wiezel Patricia Tiemi Fujimura Carlos Ueira-Vieira Eliane Candiani Arantes |
| author_sort | Priscila Yumi Tanaka Shibao |
| collection | DOAJ |
| description | Abstract Background Animal 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. Methods Total 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. Results We were able to identify several classes of antimicrobial peptides, such as buforins, peroniins and brevinins, as well as PLA2, lectins and galectins, combining protein sequencing and RNA-seq analysis for the first time. In addition, we could isolate a PLA2 from the skin secretion and infer the presence of serine proteases in cutaneous secretion. Conclusions We 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. |
| first_indexed | 2024-12-13T10:48:30Z |
| format | Article |
| id | doaj.art-66d4c9e3485740318457ac03d2366a44 |
| institution | Directory Open Access Journal |
| issn | 1678-9199 |
| language | English |
| last_indexed | 2024-12-13T10:48:30Z |
| publishDate | 2018-11-01 |
| publisher | SciELO |
| record_format | Article |
| series | Journal of Venomous Animals and Toxins including Tropical Diseases |
| spelling | doaj.art-66d4c9e3485740318457ac03d2366a442022-12-21T23:50:00ZengSciELOJournal of Venomous Animals and Toxins including Tropical Diseases1678-91992018-11-0124111510.1186/s40409-018-0173-8Deep sequencing analysis of toad Rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretionPriscila Yumi Tanaka Shibao0Camila Takeno Cologna1Romualdo Morandi-Filho2Gisele Adriano Wiezel3Patricia Tiemi Fujimura4Carlos Ueira-Vieira5Eliane Candiani Arantes6Laboratory of Animal Toxins, School of Pharmaceutical Scienes of Ribeirão Preto, University of São PauloLaboratory of Animal Toxins, School of Pharmaceutical Scienes of Ribeirão Preto, University of São PauloLaboratory of genetics – LABGEN, Institute of Genetics and Biochemistry, Campus Umuarama, Federal University of UberlândiaLaboratory of Animal Toxins, School of Pharmaceutical Scienes of Ribeirão Preto, University of São PauloLaboratory of genetics – LABGEN, Institute of Genetics and Biochemistry, Campus Umuarama, Federal University of UberlândiaLaboratory of genetics – LABGEN, Institute of Genetics and Biochemistry, Campus Umuarama, Federal University of UberlândiaLaboratory of genetics – LABGEN, Institute of Genetics and Biochemistry, Campus Umuarama, Federal University of UberlândiaAbstract Background Animal 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. Methods Total 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. Results We were able to identify several classes of antimicrobial peptides, such as buforins, peroniins and brevinins, as well as PLA2, lectins and galectins, combining protein sequencing and RNA-seq analysis for the first time. In addition, we could isolate a PLA2 from the skin secretion and infer the presence of serine proteases in cutaneous secretion. Conclusions We 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.http://link.springer.com/article/10.1186/s40409-018-0173-8RNA-seqRhinella schneideriToad secretionTranscriptomeIlluminaCutaneous secretion |
| spellingShingle | Priscila Yumi Tanaka Shibao Camila Takeno Cologna Romualdo Morandi-Filho Gisele Adriano Wiezel Patricia Tiemi Fujimura Carlos Ueira-Vieira Eliane Candiani Arantes Deep sequencing analysis of toad Rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretion Journal of Venomous Animals and Toxins including Tropical Diseases RNA-seq Rhinella schneideri Toad secretion Transcriptome Illumina Cutaneous secretion |
| title | Deep sequencing analysis of toad Rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretion |
| title_full | Deep sequencing analysis of toad Rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretion |
| title_fullStr | Deep sequencing analysis of toad Rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretion |
| title_full_unstemmed | Deep sequencing analysis of toad Rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretion |
| title_short | Deep sequencing analysis of toad Rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretion |
| title_sort | deep sequencing analysis of toad rhinella schneideri skin glands and partial biochemical characterization of its cutaneous secretion |
| topic | RNA-seq Rhinella schneideri Toad secretion Transcriptome Illumina Cutaneous secretion |
| url | http://link.springer.com/article/10.1186/s40409-018-0173-8 |
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