Evidence-Tiered Venomics of the BotG50 Toxic Fraction from Buthus tunetanus Reveals a Diversified Neurotoxin Repertoire
4 affiliations
- 1Laboratory of Venoms and Therapeutic Biomolecules, Institut Pasteur de Tunis, Université de Tunis El Manar, 13 Place Pasteur, BP74, Tunis 1002, Tunisia.
- 2Laboratory of Biomolecules, Venoms and Theranostic Applications (LBVAT), Institut Pasteur de Tunis, Université de Tunis El Manar, 13 Place Pasteur, BP74, Tunis 1002, Tunisia.
- 3Laboratoire de PhysioMédecine Moléculaire (LP2M), UMR 7370, CNRS, Université Côte d'Azur, 06107 Nice, France.
- 4Laboratory Transporter in Imaging and Radiotherapy in Oncology (TIRO), Direction de la Recherche Fondamentale (DRF), Institut des Sciences du Vivant Frédéric Joliot, Commissariat à l'Énergie Atomique et aux Énergies Alterna-tives (CEA), Université Côte d'Azur, 06107 Nice, France.
Abstract
Scorpion venoms represent complex pharmacological arsenals whose complete molecular inventory remains incompletely defined, particularly for North African buthid species of high medical relevance. Buthus tunetanus is among the most clinically significant scorpion species across the Maghreb. We analysed the toxic fraction BotG50, isolated by Sephadex G-50 gel-filtration, using nanoLC-HRMS/MS coupled with an evidence-tiered, five-stage bioinformatics workflow. The workflow combines Proteome Discoverer database searching, BLAST-guided homology expansion, Casanovo de novo sequencing with variant construction, signal-peptide and propeptide removal, and post-translational modification (PTM) annotation. The analysis revealed 64 components (FDR < 1%), comprising 38 multi-peptide-supported identifications (≥2 peptides) and 26 single-peptide hits, with a median sequence coverage of 33%. The dominant neurotoxic repertoire includes toxins targeting K+, Na+, and Cl- channels, α/β toxins, insect- or mammal-specific neurotoxins, and putative depressant toxins, while a minor accessory compartment comprises hyaluronidase, phospholipase A2, lipolytic peptides, and hemocyanin-like proteins. BLAST analysis revealed strong sequence homologies with peptides from Buthus israelis (32.8%), B. tunetanus (15.6%), and Mesobuthus eupeus (10.9%). Approximately 52% of final identifications (33/64 proteins) required analytical stages beyond classical FASTA database searching. This proteomic atlas provides the most detailed molecular map of BotG50 to date, constituting a critical resource for antivenom epitope design, nanobody target selection, and bioactive peptide discovery.
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BibTeX
@article{elfessi2026evidencetiered,
title = {{Evidence-Tiered Venomics of the BotG50 Toxic Fraction from Buthus tunetanus Reveals a Diversified Neurotoxin Repertoire}},
author = {Rym ElFessi and Rahma Ben Abderrazek and Urvashi Khurana and Thierry Pourcher and Riadh Kharrat and Saïd Bendahhou and Riadh Marrouchi},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27198876},
url = {https://hal.science/hal-05828714v1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Evidence-Tiered Venomics of the BotG50 Toxic Fraction from Buthus tunetanus Reveals a Diversified Neurotoxin Repertoire
AU - Rym ElFessi
AU - Rahma Ben Abderrazek
AU - Urvashi Khurana
AU - Thierry Pourcher
AU - Riadh Kharrat
AU - Saïd Bendahhou
AU - Riadh Marrouchi
PY - 2026
DA - 2026/10/06
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27198876
UR - https://hal.science/hal-05828714v1
AB - Scorpion venoms represent complex pharmacological arsenals whose complete molecular inventory remains incompletely defined, particularly for North African buthid species of high medical relevance. Buthus tunetanus is among the most clinically significant scorpion species across the Maghreb. We analysed the toxic fraction BotG50, isolated by Sephadex G-50 gel-filtration, using nanoLC-HRMS/MS coupled with an evidence-tiered, five-stage bioinformatics workflow. The workflow combines Proteome Discoverer database searching, BLAST-guided homology expansion, Casanovo de novo sequencing with variant construction, signal-peptide and propeptide removal, and post-translational modification (PTM) annotation. The analysis revealed 64 components (FDR < 1%), comprising 38 multi-peptide-supported identifications (≥2 peptides) and 26 single-peptide hits, with a median sequence coverage of 33%. The dominant neurotoxic repertoire includes toxins targeting K+, Na+, and Cl- channels, α/β toxins, insect- or mammal-specific neurotoxins, and putative depressant toxins, while a minor accessory compartment comprises hyaluronidase, phospholipase A2, lipolytic peptides, and hemocyanin-like proteins. BLAST analysis revealed strong sequence homologies with peptides from Buthus israelis (32.8%), B. tunetanus (15.6%), and Mesobuthus eupeus (10.9%). Approximately 52% of final identifications (33/64 proteins) required analytical stages beyond classical FASTA database searching. This proteomic atlas provides the most detailed molecular map of BotG50 to date, constituting a critical resource for antivenom epitope design, nanobody target selection, and bioactive peptide discovery.
SN - 1661-6596
SN - 1422-0067
ER -