{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T05:08:04Z","timestamp":1768280884272,"version":"3.49.0"},"reference-count":66,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2022,11,22]],"date-time":"2022-11-22T00:00:00Z","timestamp":1669075200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["AI139306"],"award-info":[{"award-number":["AI139306"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Botulinum neurotoxin serotype A (BoNT\/A) is the most potent protein toxin for humans and is utilized as a therapy for numerous neurologic diseases. BoNT\/A comprises a catalytic Light Chain (LC\/A) and a Heavy Chain (HC\/A) and includes eight subtypes (BoNT\/A1-\/A8). Previously we showed BoNT\/A potency positively correlated with stable localization on the intracellular plasma membrane and identified a low homology domain (amino acids 268\u2013357) responsible for LC\/A1 stable co-localization with SNAP-25 on the plasma membrane, while LC\/A3 was present in the cytosol of Neuro2A cells. In the present study, steady-state- and live-imaging of a cytosolic LC\/A3 derivative (LC\/A3V) engineered to contain individual structural elements of the A1 LDH showed that a 59 amino acid region (275\u2013334) termed the MLD was sufficient to direct LC\/A3V from the cytosol to the plasma membrane co-localized with SNAP-25. Informatics and experimental validation of the MLD-predicted R1 region (an \u03b1-helix, residues 275\u2013300) and R2 region (a loop, \u03b1-helix, loop, residues 302\u2013334) both contribute independent steps to the stable co-localization of LC\/A1 with SNAP-25 on the plasma membrane of Neuro-2A cells. Understanding how these structural elements contribute to the overall association of LC\/A1 on the plasma membrane may identify the molecular basis for the LC contribution of BoNT\/A1 to high potency.<\/jats:p>","DOI":"10.3390\/toxins14120814","type":"journal-article","created":{"date-parts":[[2022,11,22]],"date-time":"2022-11-22T05:18:57Z","timestamp":1669094337000},"page":"814","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["How Botulinum Neurotoxin Light Chain A1 Maintains Stable Association with the Intracellular Neuronal Plasma Membrane"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4503-1145","authenticated-orcid":false,"given":"Alexander P.","family":"Gardner","sequence":"first","affiliation":[{"name":"Microbiology and Immunology, Medical College, Wisconsin 8701 Watertown Plank Road, Milwaukee, WI 53226, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph T.","family":"Barbieri","sequence":"additional","affiliation":[{"name":"Microbiology and Immunology, Medical College, Wisconsin 8701 Watertown Plank Road, Milwaukee, WI 53226, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8186-8810","authenticated-orcid":false,"given":"Sabine","family":"Pellett","sequence":"additional","affiliation":[{"name":"Department of Bacteriology, Microbial Sciences Building, University of Wisconsin-Madison, 1550 Linden Dr., Madison, WI 53706, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"469","DOI":"10.3390\/toxins3050469","article-title":"Botulinum neurotoxins and botulism: A novel therapeutic approach","volume":"3","author":"Thanongsaksrikul","year":"2011","journal-title":"Toxins"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1152\/physrev.2000.80.2.717","article-title":"Neurotoxins affecting neuroexocytosis","volume":"80","author":"Schiavo","year":"2000","journal-title":"Physiol. 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