{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,29]],"date-time":"2025-08-29T10:14:27Z","timestamp":1756462467106},"reference-count":50,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2019,4,17]],"date-time":"2019-04-17T00:00:00Z","timestamp":1555459200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["ZIA AI000929\u201015"],"award-info":[{"award-number":["ZIA AI000929\u201015"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000072","name":"National Institute of Dental and Craniofacial Research","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000072","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2019,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:label \/><jats:p>In recent years, anthrax toxin has been reengineered to act as a highly specific antiangiogenic cancer therapeutic, shown to kill tumors in animal models. This has been achieved by modifying protective antigen (PA) so that its activation and toxicity require the presence of two proteases, matrix metalloproteinase (MMP) and urokinase plasminogen activator (uPA), which are upregulated in tumor microenvironments. These therapeutics consist of intercomplementing PA variants, which are individually nontoxic, but form functional toxins upon complementary oligomerization. Here, we have created a dual\u2010protease requiring PA targeting system which utilizes bismaleimide cross\u2010linked PA (CLPA) rather than the intercomplementing PA variants. Three different CLPA agents were tested and, as expected, found to exclusively form octamers. Two of the CLPA agents have <jats:italic>in vitro<\/jats:italic> toxicities equal to those of previous intercomplementing agents, while the third CLPA agent had compromised <jats:italic>in vitro<\/jats:italic> cleavage and was significantly less cytotoxic. We hypothesize this difference was due to steric hindrance caused by cross\u2010linking two PA monomers in close proximity to the PA cleavage site. Overall, this work advances the development and use of the PA and LF tumor\u2010targeting system as a practical cancer therapeutic, as it provides a way to reduce the drug components of the anthrax toxin drug delivery system from three to two, which may lower the cost and simplify testing in clinical trials.<\/jats:p><\/jats:sec><jats:sec><jats:title>HIGHLIGHT<\/jats:title><jats:p>Previously, anthrax toxin has been reengineered to act as a highly specific antiangiogenic cancer therapeutic. Here, we present a version, which utilizes bismaleimide cross\u2010linked protective antigen (PA) rather than intercomplementing PA variants. This advances the development of anthrax toxin as a practical cancer therapeutic as it reduces the components of the drug delivery system to two, which may lower the cost and simplify testing in clinical trials.<\/jats:p><\/jats:sec>","DOI":"10.1002\/pro.3613","type":"journal-article","created":{"date-parts":[[2019,4,3]],"date-time":"2019-04-03T15:28:12Z","timestamp":1554305292000},"page":"1059-1070","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Bismaleimide cross\u2010linked anthrax toxin forms functional octamers with high specificity in tumor targeting"],"prefix":"10.1002","volume":"28","author":[{"given":"Elyse S.","family":"Fischer","sequence":"first","affiliation":[{"name":"Laboratory of Parasitic Diseases National Institute of Allergy and Infectious Diseases  Bethesda Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"suffix":"IV","given":"Warren 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Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas H.","family":"Bugge","sequence":"additional","affiliation":[{"name":"Oral and Pharyngeal Cancer Branch National Institute of Dental and Craniofacial Research  Bethesda Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen H.","family":"Leppla","sequence":"additional","affiliation":[{"name":"Laboratory of Parasitic Diseases National Institute of Allergy and Infectious Diseases  Bethesda 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