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. 2017 Jul 11:4:170090.
doi: 10.1038/sdata.2017.90.

A multi-protease, multi-dissociation, bottom-up-to-top-down proteomic view of the Loxosceles intermedia venom

Dilza Trevisan-Silva  1 , Aline V Bednaski  1 , Juliana S G Fischer  2 , Silvio S Veiga  1 , Nuno Bandeira  3 , Adrian Guthals  3 , Fabricio K Marchini  4   5 , Felipe V Leprevost  2 , Valmir C Barbosa  6 , Andrea Senff-Ribeiro  1 , Paulo C Carvalho  2   7
Affiliations

A multi-protease, multi-dissociation, bottom-up-to-top-down proteomic view of the Loxosceles intermedia venom

Dilza Trevisan-Silva et al. Sci Data. .

Abstract

Venoms are a rich source for the discovery of molecules with biotechnological applications, but their analysis is challenging even for state-of-the-art proteomics. Here we report on a large-scale proteomic assessment of the venom of Loxosceles intermedia, the so-called brown spider. Venom was extracted from 200 spiders and fractioned into two aliquots relative to a 10 kDa cutoff mass. Each of these was further fractioned and digested with trypsin (4 h), trypsin (18 h), pepsin (18 h), and chymotrypsin (18 h), then analyzed by MudPIT on an LTQ-Orbitrap XL ETD mass spectrometer fragmenting precursors by CID, HCD, and ETD. Aliquots of undigested samples were also analyzed. Our experimental design allowed us to apply spectral networks, thus enabling us to obtain meta-contig assemblies, and consequently de novo sequencing of practically complete proteins, culminating in a deep proteome assessment of the venom. Data are available via ProteomeXchange, with identifier PXD005523.

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Conflict of interest statement

N.B. has an equity interest in Digital Proteomics, LLC, a company that may potentially benefit from the research results; Digital Proteomics, LLC was not involved in any aspects of this research. The terms of this arrangement have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. The remaining authors declares no competing financial interests.

Figures

Figure 1
Figure 1. Methodology workflow.
Summary of the sequence of procedures that constitute the methodology employed, from venom extraction to the meta-contig assembling that enabled the identifications of venom proteins.
See this image and copyright information in PMC

References

Data Citations

    1. Trevisan-Silva D. 2016. Dummy. PXD005523
    1. Trevisan-Silva D. 2017. Figshare. https://doi.org/10.6084/m9.figshare.c.3709168 - DOI

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