{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:06:52Z","timestamp":1792789612223,"version":"4.5.0"},"reference-count":55,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Pasteur Institute of Tunis"},{"name":"Ministry of Health"},{"name":"Ministry of Higher Education"},{"name":"Scientific Research of Tunisia"},{"name":"Universit\u00e9 C\u00f4te d\u2019Azur","id":[{"id":"https:\/\/ror.org\/019tgvf94","id-type":"ROR","asserted-by":"crossref"}]},{"id":[{"id":"https:\/\/ror.org\/019tgvf94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>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 &lt; 1%), comprising 38 multi-peptide-supported identifications (\u22652 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, \u03b1\/\u03b2 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.<\/jats:p>","DOI":"10.3390\/ijms27198876","type":"journal-article","created":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T09:34:18Z","timestamp":1791279258000},"page":"8876","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Evidence-Tiered Venomics of the BotG50 Toxic Fraction from Buthus tunetanus Reveals a Diversified Neurotoxin Repertoire"],"prefix":"10.3390","volume":"27","author":[{"given":"Rym","family":"ElFessi","sequence":"first","affiliation":[{"name":"Laboratory of Venoms and Therapeutic Biomolecules, Institut Pasteur de Tunis, Universit\u00e9 de Tunis El Manar, 13 Place Pasteur, BP74, Tunis 1002, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rahma Ben","family":"Abderrazek","sequence":"additional","affiliation":[{"name":"Laboratory of Biomolecules, Venoms and Theranostic Applications (LBVAT), Institut Pasteur de Tunis, Universit\u00e9 de Tunis El Manar, 13 Place Pasteur, BP74, Tunis 1002, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Urvashi","family":"Khurana","sequence":"additional","affiliation":[{"name":"Laboratoire de PhysioM\u00e9decine Mol\u00e9culaire (LP2M), UMR 7370, CNRS, Universit\u00e9 C\u00f4te d\u2019Azur, 06107 Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2839-4622","authenticated-orcid":false,"given":"Thierry","family":"Pourcher","sequence":"additional","affiliation":[{"name":"Laboratory Transporter in Imaging and Radiotherapy in Oncology (TIRO), Direction de la Recherche Fondamentale (DRF), Institut des Sciences du Vivant Fr\u00e9d\u00e9ric Joliot, Commissariat \u00e0 l\u2019\u00c9nergie Atomique et aux \u00c9nergies Alterna-tives (CEA), Universit\u00e9 C\u00f4te d\u2019Azur, 06107 Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Riadh","family":"Kharrat","sequence":"additional","affiliation":[{"name":"Laboratory of Venoms and Therapeutic Biomolecules, Institut Pasteur de Tunis, Universit\u00e9 de Tunis El Manar, 13 Place Pasteur, BP74, Tunis 1002, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sa\u00efd","family":"Bendahhou","sequence":"additional","affiliation":[{"name":"Laboratoire de PhysioM\u00e9decine Mol\u00e9culaire (LP2M), UMR 7370, CNRS, Universit\u00e9 C\u00f4te d\u2019Azur, 06107 Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2311-576X","authenticated-orcid":false,"given":"Riadh","family":"Marrouchi","sequence":"additional","affiliation":[{"name":"Laboratory of Venoms and Therapeutic Biomolecules, Institut Pasteur de Tunis, Universit\u00e9 de Tunis El Manar, 13 Place Pasteur, BP74, Tunis 1002, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,10,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Huang, Y., Kamau, P.M., Wang, J., Gao, M., and Li, B. 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