{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T20:06:53Z","timestamp":1771963613247,"version":"3.50.1"},"reference-count":66,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"35","license":[{"start":{"date-parts":[[2025,8,27]],"date-time":"2025-08-27T00:00:00Z","timestamp":1756252800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.science.org\/content\/page\/science-licenses-journal-article-reuse"}],"content-domain":{"domain":["www.science.org"],"crossmark-restriction":true},"short-container-title":["Sci. Adv."],"published-print":{"date-parts":[[2025,8,29]]},"abstract":"<jats:p>Botulinum neurotoxin serotype A (BoNT\/A) is naturally produced by bacteria along with four nontoxic neurotoxin-associated proteins (NTNH, HA70, HA33, and HA17), forming a bimodular large progenitor toxin complex (L-PTC). The BoNT\/A\u2013NTNH complex protects the toxin from adverse environment, while the complex consisting of HA proteins facilitates toxin absorption during oral intoxication. How these two independent modules assemble into the L-PTC remains unclear. Here, we report the crystal structure of the BoNT\/A\u2013NTNH\u2013HA70 complex at ~2.9-\u00c5 resolution. The structure reveals that the BoNT\/A\u2013NTNH complex is anchored into a concentric double \u03b2-barrel channel of trimeric HA70 through a short \u03b2-hairpin of NTNH (termed nLoop), resembling a nut-and-bolt attachment. We find that the nLoop of NTNH is strictly conserved across HA-containing BoNT complexes and that NTNH\u2013HA70 binding is interchangeable among them. Furthermore, we demonstrate that the nLoop functions as a minimal motif enabling attachment of a protein-of-interest to the HA complex, with potential applications in oral biologics delivery.<\/jats:p>","DOI":"10.1126\/sciadv.adx5831","type":"journal-article","created":{"date-parts":[[2025,8,27]],"date-time":"2025-08-27T17:59:06Z","timestamp":1756317546000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":2,"title":["A nut-and-bolt assembly of the bimodular large progenitor botulinum neurotoxin complex"],"prefix":"10.1126","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7173-6600","authenticated-orcid":true,"given":"Kwok-ho","family":"Lam","sequence":"first","affiliation":[{"name":"Department of Physiology and Biophysics, University of California, Irvine, CA 92697, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6194-5145","authenticated-orcid":true,"given":"Linfeng","family":"Gao","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, University of California, Irvine, CA 92697, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adina","family":"Przykopanski","sequence":"additional","affiliation":[{"name":"Institute for Toxicology, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8859-2762","authenticated-orcid":true,"given":"Baohua","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, University of California, Irvine, CA 92697, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ting","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, University of California, Irvine, CA 92697, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9775-3110","authenticated-orcid":true,"given":"Maren","family":"Kr\u00fcger","sequence":"additional","affiliation":[{"name":"Biological Toxins, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Seestrasse 10, 13353 Berlin, Germany."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna-Magdalena","family":"Bartels","sequence":"additional","affiliation":[{"name":"Institute for Toxicology, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8176-797X","authenticated-orcid":true,"given":"Martin Bernhard","family":"Dorner","sequence":"additional","affiliation":[{"name":"Biological Toxins, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Seestrasse 10, 13353 Berlin, Germany."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4046-1704","authenticated-orcid":true,"given":"Kay","family":"Perry","sequence":"additional","affiliation":[{"name":"NE-CAT and Department of Chemistry and Chemical Biology, Cornell University, Argonne National Laboratory, Argonne, IL 60439, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5100-2546","authenticated-orcid":true,"given":"Brigitte Gertrud","family":"Dorner","sequence":"additional","affiliation":[{"name":"Biological Toxins, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Seestrasse 10, 13353 Berlin, Germany."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0645-1641","authenticated-orcid":true,"given":"Andreas","family":"Rummel","sequence":"additional","affiliation":[{"name":"Institute for Toxicology, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0348-7363","authenticated-orcid":true,"given":"Rongsheng","family":"Jin","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, University of California, Irvine, CA 92697, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"221","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro3295"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.3390\/toxins9010038"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1186\/1741-7007-7-66"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1214270"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2015.03.013"},{"key":"e_1_3_2_7_2","first-page":"21","article-title":"Assembly and function of the botulinum neurotoxin progenitor complex","volume":"364","author":"Gu S.","year":"2013","unstructured":"S. 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