Revealing the functional structure of a new PLA2 K49 from Bothriopsis taeniata snake venom employing automatic “de novo” sequencing using CID/HCD/ETD MS/MS analyses
peer-reviewed · Journal of Proteomics · 2015
| Date | 2015-10-22 |
| Type | peer-reviewed |
| Venue | Journal of Proteomics |
| Publisher | Elsevier BV |
| Contribution | downstream-application |
| DOI | 10.1016/j.jprot.2015.10.020 |
| Citations (OpenAlex) | 11 |
| Venue 2-year citedness | 3.02 |
Abstract
Snake venoms are composed of approximately 90% of proteins with several pharmacological activities having high potential in research as biological tools. One of the most abundant compounds is phospholipases A2 (PLA2), which are the most studied venom protein due to their wide pharmacological activity. Using a combination of chromatographic steps, a new PLA2 K49 was isolated and purified from the whole venom of the Bothriopsis taeniata and submitted to analyses mass spectrometry. An automatic “de novo” sequencing of this new PLA2 K49 denominated Btt-TX was performed using Peaks Studio 6 for analysis of the spectra. Additionally, a triplex approach CID/HCD/ETD has been performed, to generate higher coverage of the sequence of the protein. Structural studies correlating biological activities were made associating specific Btt-TX regions and myotoxic activity. Lysine acetylation was performed to better understand the mechanism of membrane interaction, identifying the extreme importance of the highly hydrophobic amino acids L, P and F for disruption of the membrane. Our myotoxical studies show a possible membrane disruption mechanism by Creatine Kinase release without a noticeable muscle damage, that probably occurred without phospholipid hydrolyses, but with a probable penetration of the hydrophobic amino acids present in the C-terminal region of the protein.
Methods and tools
- Bothriopsis taeniata PLA2 K49: Automatic de novo sequencing (PEAKS Studio) with CID/HCD/ETD triplex MS/MS to elucidate the full-length sequence of a new PLA2 K49 (Btt-TX) from Bothriopsis taeniata snake venom.
Methods it uses
- PEAKS: Commercial DP-based de novo
Cites (6)
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- Constrained De Novo Sequencing of Conotoxins (2012) crossref
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- Shotgun Protein Sequencing: Assembly of Peptide Tandem Mass Spectra from Mixtures of Modified Proteins (2007) crossref
- De Novo Peptide Sequencing and Identification with Precision Mass Spectrometry (2007) crossref
- Shotgun Protein Sequencing by Tandem Mass Spectra Assembly (2004) crossref