Dataset Information

Comparative analysis of high molecular mass subproteomes of Bothrops snake venoms

ABSTRACT: Snake venoms are extremely active biological secretions containing primarily various classes of enzymes. The genus Bothrops comprises various pit viper species that represent the most medically significant taxa in Central and South America, accounting for more human envenomations and fatalities than any other snake taxa. Venom proteomes of many Bothrops species have been characterized but, although proteins of molecular mass higher than 100 kDa have been documented in Bothrops venoms, these large proteins have not yet been closely investigated. This study sought to achieve detailed identification of major components in the high molecular mass subproteome of venoms from eight Bothrops species (B. brazili, B. cotiara, B. insularis, B. jararaca, B. jararacussu, B. leucurus, B. moojeni and B. neuwiedi). The identification of proteins eluting in the first fractions of a size-exclusion chromatography of these venoms revealed that they are comprised mainly of enzymes, including minor components such as 5'-nucleotidase, aminopeptidase, phosphodiesterase, and phospholipases A2 and B, but with metalloproteinases and L-amino acid oxidases representing the most abundant components. Most of these components disappeared in electrophoretic profiles under reducing conditions suggesting that they may be composed of more than one polypeptide chain. A significant shift in the molecular masses of these protein bands was observed after enzymatic N-deglycosylation, indicating that they may contain N-glycans. Furthermore, the finding that none of the high molecular mass proteins is shared by all eight species reveals a high level of interspecies venom variability among these components.

INSTRUMENT(S): Q-Tof Ultima, LTQ Orbitrap Velos

ORGANISM(S): Acyrthosiphon Pisum (pea Aphid) Bothrops Cotiara Bothrops Jararacussu Bothrops Leucurus Bothrops Brazili Bothrops Neuwiedi Bothrops Insularis Bothrops Jararaca (jararaca) (bothrops Jajaraca) Bothrops Moojeni

SUBMITTER: Solange Serrrano

LAB HEAD: Solange Maria de Toledo Serrano

PROVIDER: PXD012088 | Pride | 2019-03-05

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	OB_EG_INSOLUTION_mgf.zip	Other
	OB_EG_INSOLUTION_raw.zip	Other
	Q_EG_BJARARACA2_BANDS1_8_20160328_RAW.zip	Other
	Q_EG_BJARARACA2_BANDS_1_8.pkl_files.zip	Other
	Q_EG_BJARARACUSSU_BANDS_1_7.pkl_files.zip	Other

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Publications

Comparative analysis of the high molecular mass subproteomes of eight Bothrops snake venoms.

Gren Eric C K ECK Kitano Eduardo S ES Andrade-Silva Débora D Iwai Leo Kei LK Reis Marcelo S MS Menezes Milene C MC Serrano Solange M T SMT

Comparative biochemistry and physiology. Part D, Genomics & proteomics 20190130

Snake venoms are extremely active biological secretions composed primarily of various classes of enzymes. The genus Bothrops comprises various pit viper species that represent the most medically significant taxa in Central and South America, accounting for more human envenomations and fatalities than any other snakes in the region. Venom proteomes of many Bothrops species have been well-characterized but investigations have focused almost exclusively on proteins smaller than 100 kDa despite expr ...[more]

PMID: 30825662

Similar Datasets

Project description:Snake venoms are complex protein mixtures with different biological activities that can act in both their preys and human victims. Many of these proteins play a role in prey capture and in the digestive process of these animals. It is known that some snakes are resistant to the toxicity of their own venom by mechanisms not yet fully elucidated. However, it was observed in the Laboratory of Herpetology of Instituto Butantan that some Bothrops moojeni individuals injured by the same snake species showed mortalities caused by envenoming effects. This study analyzed the biochemical composition of 13 venom and plasma samples from Bothrops moojeni specimens to assess differences in their protein composition. Application of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed distinct venom protein profiles, but very homogeneous plasma profiles. Western Blotting (WB) was performed with plasma samples, which were submitted to incubation with the respective venom. Some individuals showed an immunorecognized band zone around 25 kDa, indicating interaction between the same individual plasma and venom proteins. Crossed-WB assay using non-self-plasma and venom showed that this variability is due to venom protein composition instead of plasma composition. These venoms presented higher caseinolytic, collagenolytic and coagulant activities than the venoms without these regions recognized by WB. Mass spectrometry analysis revealed that these individuals present, in addition to higher protein concentrations, other exclusive proteins in their composition. When these samples were tested in vivo, the results also showed higher lethality in these venoms, but lower hemorrhagic activity than in the venoms without these regions recognized by WB. In conclusion, some Bothrops moojeni specimens differ in venom composition, which may have implications in envenomation. Moreover, the high individual venom variability found in this species demonstrates the importance to work with individual analysis in studies involving intraspecific venom variability and venom evolution.

Project description:Spider venoms are a unique source of bioactive peptides displaying remarkable structural stability and potent neuroactivity. Phoneutria nigriventer, often referred to as Brazilian wandering spider, banana spider or “armed” spider, is endemic from South America and amongst the most dangerous venomous spiders in the world. Envenomation accidents with P. nigriventer often occur in Brazil with approximately 4,000 cases per year and which symptoms include priapism, hypertension, blurred vision, sweating, and vomiting, amongst others. Besides its clinical relevance, P. nigriventer venom comprises promising peptide drug leads providing therapeutic effects in a range of disease models. In this study, we further explored the neuroactivity and molecular diversity of the venom from P. nigriventer using fractionation-guided high-throughput cellular assays coupled to proteomics analysis and multi-pharmacology activity to broaden the knowledge and therapeutic potential of this venom, as well as a proof-of-concept for an investigative pipeline to study spider-venom derived neuroactive peptides. We applied ion channel assays in a neuroblastoma cell line to investigate modulation of voltage-gated sodium and calcium channels, and nicotinic acetylcholine α7 receptor. We then investigated the venom components using mass spectrometry to identify peptide masses and sequences associated to the observed neuromodulations. Our findings showed this venom is highly complex compared to other neurotoxin-rich venoms and comprises potent modulators of voltage-gated sodium and calcium channels which were classified into 4 families of neuroactive peptides based on their activities and structures. In addition to the reported P. nigriventer neuroactive peptides, we identified at least 27 novel cysteine-rich venom peptides in which neuroactivities are still to be elucidated in voltage-gated ion channels and other potential targets. Our findings provide a new basis for studying non-explored bioactivities of known and novel neuroactive components in P. nigriventer venom, and further supports the successful application of our discovery pipeline for identifying ion channel-targeting venom peptides with potential to become drug leads with diverse exploratory and therapeutic applications.