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1.
Protein J ; 36(2): 77-97, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-28258523

RESUMO

Pelagia noctiluca is the most venomous jellyfish in the Mediterranean Sea where it forms dense blooms. Although there is several published research on this species, until now none of the works has been focused on a complete protein profile of the all body constituents of this organism. Here, we have performed a detailed proteomics characterization of the major protein components expressed by P. noctiluca. With that aim, we have considered the study of jellyfish proteins involved in defense, body constituents and metabolism, and furthered explore the significance and potential application of such bioactive molecules. P. noctiluca body proteins were separated by1D SDS-PAGE and 2DE followed by characterization by nanoLC-MS/MS and MALDI-TOF/TOF techniques. Altogether, both methods revealed 68 different proteins, including a Zinc Metalloproteinase, a Red Fluorescent Protein (RFP) and a Peroxiredoxin. These three proteins were identified for the first time in P. noctiluca. Zinc Metalloproteinase was previously reported in the venom of other jellyfish species. Besides the proteins described above, the other 65 proteins found in P. noctiluca body content were identified and associated with its clinical significance. Among all the proteins identified in this work we highlight: Zinc metalloproteinase, which has a ShK toxin domain and therefore should be implicated in the sting toxicity of P. noctiluca.; the RFP which are a very important family of proteins due to its possible application as molecular markers; and last but not least the discovery of a Peroxiredoxin in this organism makes it a new natural resource of antioxidant and anti-UV radiation agents.


Assuntos
Proteínas Luminescentes/análise , Metaloproteases/análise , Peroxirredoxinas/análise , Proteoma/análise , Cifozoários/metabolismo , Animais , Venenos de Cnidários/análise , Venenos de Cnidários/química , Eletroforese , Proteínas Luminescentes/química , Proteínas Luminescentes/metabolismo , Mar Mediterrâneo , Metaloproteases/química , Metaloproteases/metabolismo , Peroxirredoxinas/química , Peroxirredoxinas/metabolismo , Domínios Proteicos , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem , Zinco , Proteína Vermelha Fluorescente
2.
Mar Drugs ; 14(4)2016 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-27077869

RESUMO

The study of bioactive compounds from marine animals has provided, over time, an endless source of interesting molecules. Jellyfish are commonly targets of study due to their toxic proteins. However, there is a gap in reviewing successful wet-lab methods employed in these animals, which compromises the fast progress in the detection of related biomolecules. Here, we provide a compilation of the most effective wet-lab methodologies for jellyfish venom extraction prior to proteomic analysis-separation, identification and toxicity assays. This includes SDS-PAGE, 2DE, gel chromatography, HPLC, DEAE, LC-MS, MALDI, Western blot, hemolytic assay, antimicrobial assay and protease activity assay. For a more comprehensive approach, jellyfish toxicity studies should further consider transcriptome sequencing. We reviewed such methodologies and other genomic techniques used prior to the deep sequencing of transcripts, including RNA extraction, construction of cDNA libraries and RACE. Overall, we provide an overview of the most promising methods and their successful implementation for optimizing time and effort when studying jellyfish.


Assuntos
Fatores Biológicos/genética , Fatores Biológicos/metabolismo , Venenos de Cnidários/genética , Genômica/métodos , Proteômica/métodos , Cifozoários/genética , Animais , Biblioteca Gênica , Humanos
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