{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T16:21:38Z","timestamp":1791044498892,"version":"4.1.0"},"reference-count":32,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2020,5,14]],"date-time":"2020-05-14T00:00:00Z","timestamp":1589414400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbiology and Immunology"],"published-print":{"date-parts":[[2020,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Botulinum neurotoxin (BoNT) is the causative agent of botulism in humans and animals. Only BoNT serotype A subtype 1 (BoNT\/A1) is used clinically because of its high potency and long duration of action. BoNT\/A1 and BoNT\/A subtype 2 (BoNT\/A2) have a high degree of amino acid sequence similarity in the light chain (LC) (96%), whereas their N\u2010and C\u2010terminal heavy chain (H<jats:sub>N<\/jats:sub> and H<jats:sub>C<\/jats:sub>) differ by 13%. The LC acts as a zinc\u2010dependent endopeptidase, H<jats:sub>N<\/jats:sub> as the translocation domain, and H<jats:sub>C<\/jats:sub> as the receptor\u2010binding domain. BoNT\/A2 and BoNT\/A1 had similar potency in the mouse bioassay, but BoNT\/A2 entered faster and more efficiently into neuronal cells. To identify the domains responsible for these characteristics, H<jats:sub>N<\/jats:sub> of BoNT\/A1 and BoNT\/A2 was exchanged to construct chimeric BoNT\/A121 and BoNT\/A212. After expression in <jats:italic>Escherichia coli<\/jats:italic>, chimeric and wild\u2010type BoNT\/As were purified as single\u2010chain proteins and activated by conversion to disulfide\u2010linked dichains. The toxicities of recombinant wild\u2010type and chimeric BoNT\/As were similar, but dropped to 60% compared with the values of native BoNT\/As. The relative orders of SNAP\u201025 cleavage activity in neuronal cells and toxicity differed. BoNT\/A121 and recombinant BoNT\/A2 have similar SNAP\u201025 cleavage activity. BoNT\/A2 H<jats:sub>N<\/jats:sub> is possibly responsible for the higher potency of BoNT\/A2 than BoNT\/A1.<\/jats:p>","DOI":"10.1111\/1348-0421.12796","type":"journal-article","created":{"date-parts":[[2020,4,17]],"date-time":"2020-04-17T11:25:13Z","timestamp":1587122713000},"page":"502-511","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Translocation domain of botulinum neurotoxin A subtype 2 potently induces entry into neuronal cells"],"prefix":"10.1111","volume":"64","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5605-5321","authenticated-orcid":false,"given":"Tomoko","family":"Kohda","sequence":"first","affiliation":[{"name":"Department of Veterinary Science, Graduate School of Life and Environmental Sciences Osaka Prefecture University  Izumisano Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kentaro","family":"Tsukamoto","sequence":"additional","affiliation":[{"name":"Department of Microbiology Fujita Health University School of Medicine  Toyoake Aichi Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasushi","family":"Torii","sequence":"additional","affiliation":[{"name":"Department of Animal Science Tokyo University of Agriculture  Atsugi Kanagawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shunji","family":"Kozaki","sequence":"additional","affiliation":[{"name":"Department of Veterinary Science, Graduate School of Life and Environmental Sciences Osaka Prefecture University  Izumisano Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masafumi","family":"Mukamoto","sequence":"additional","affiliation":[{"name":"Department of Veterinary Science, Graduate School of Life and Environmental Sciences Osaka Prefecture University  Izumisano Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,5,14]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro3295"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2014.08.009"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.micpath.2007.12.003"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00536-13"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2010.10.009"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M513032200"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi801686b"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00795-16"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000245"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0153401"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-07789-4"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.7883\/yoken1952.37.131"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)52451-6"},{"key":"e_1_2_8_15_1","first-page":"528","article-title":"Gene splicing by overlap extension: tailor\u2010made genes using the polymerase chain reaction","volume":"8","author":"Horton RM","year":"1990","journal-title":"Biotechniques."},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF01891992"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1020687430927"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1348-0421.2012.00490.x"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.0375-08.2008"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2010.11.045"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.3389\/fneur.2014.00098"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1128\/mSphere.00466-18"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2015.09.025"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01795-14"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00089-18"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1348-0421.1995.tb03269.x"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0049516"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M507596200"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2009.07.023"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep43588"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M710442200"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1111\/1348-0421.12540"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2013.10.010"}],"container-title":["Microbiology and Immunology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2F1348-0421.12796","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1348-0421.12796","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/1348-0421.12796","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1348-0421.12796","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,6]],"date-time":"2023-09-06T06:04:00Z","timestamp":1693980240000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/1348-0421.12796"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,5,14]]},"references-count":32,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2020,7]]}},"alternative-id":["10.1111\/1348-0421.12796"],"URL":"https:\/\/doi.org\/10.1111\/1348-0421.12796","archive":["Portico"],"relation":{},"ISSN":["0385-5600","1348-0421"],"issn-type":[{"value":"0385-5600","type":"print"},{"value":"1348-0421","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,5,14]]},"assertion":[{"value":"2019-11-19","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-04-14","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-05-14","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}