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Proteomic analysis of Bothrops pirajai snake venom and characterization of BpirMP, a new P-I metalloproteinase
- Source :
- Journal of Proteomics; Volumen 80. 2013, Kérwá, Universidad de Costa Rica, instacron:UCR, Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual), Universidade de São Paulo (USP), instacron:USP
- Publication Year :
- 2013
-
Abstract
- Bothrops pirajai snake venom was analyzed by a proteomic strategy. Proteins were separated by RP-HPLC, followed by SDS-PAGE, in-gel tryptic digestion, identification by MALDI-TOF/TOF mass spectrometry, and assignment to known protein families by similarity. Proteins belonging to six families were found in B. pirajai venom, including abundant PLA 2 s and metalloproteinases, with the remaining proteins distributed among l -amino acid oxidase, serine proteinase, disintegrin and lectin-like families. A P-I class metalloproteinase, named BpirMP, was isolated from this venom by three chromatographic steps. The enzyme has a molecular mass of 23.1 kDa, as determined by mass spectrometry. Its proteolytic activity on azocasein was inhibited by chelating and reducing agents, with optimum activity at higher pH values and 37 °C. BpirMP presented weak hemorrhagic activity, with an MHD of 50 μg, and was able to hydrolyze basement membrane components in vivo and in vitro. The toxin cleaved both Aα and Bβ chains of fibrinogen and was also able to degrade fibrin and blood clots in vitro. The primary sequence analysis indicates that BpirMP contains a zinc ligand motif and a CVM motif that is associated with a Met-turn structure. These results demonstrate that BpirMP is a zinc-dependent hemorrhagic metalloproteinase with fibrin(ogen)olytic and thrombolytic activities. Biological significance This manuscript describes the diversity of protein components present in the venom of Bothops pirajai , a threatened snake species from northeastern Brazil, as well as the isolation and biochemical properties of a PI-SVMP. The results showed distinct mechanisms of action that should contribute in the elucidation of the differences in the hemorrhagic potential of SVMPs, allowing a better understanding of this class of enzymes and of the biology of Bothrops pirajai species.
- Subjects :
- Proteomics
Snake venom
Basement membrane
Proteome
Molecular Sequence Data
Biophysics
Bothrops pirajai
Hemorrhage
Venom
Viper Venoms
Biochemistry
Serine
FOSFOLIPASES A (ISOLAMENTO E PURIFICAÇÃO)
Crotalid Venoms
Disintegrin
Animals
Humans
Bothrops
Amino Acid Sequence
Metalloproteinase
chemistry.chemical_classification
biology
Molecular mass
Fibrinolysis
Fibrinogen
biology.organism_classification
Molecular biology
Amino acid
Molecular Weight
Phospholipases A2
chemistry
Metalloproteases
biology.protein
Sequence Alignment
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Journal of Proteomics; Volumen 80. 2013, Kérwá, Universidad de Costa Rica, instacron:UCR, Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual), Universidade de São Paulo (USP), instacron:USP
- Accession number :
- edsair.doi.dedup.....58a624103569e3a45d6cb1e4fbbfbcb2