{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T04:20:48Z","timestamp":1760242848772,"version":"build-2065373602"},"reference-count":39,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2016,8,18]],"date-time":"2016-08-18T00:00:00Z","timestamp":1471478400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Edema Factor (EF), the toxic sub-unit of the Bacillus anthracis Edema Toxin (ET) is a calmodulin-dependent adenylate cyclase whose detrimental activity in the infected host results in severe edema. EF is therefore a major virulence factor of B. anthracis. We describe a simple, rapid and reliable functional adenylate-cyclase assay based on inhibition of a luciferase-mediated luminescence reaction. The assay exploits the efficient adenylate cyclase-mediated depletion of adenosine tri-phosphate (ATP), and the strict dependence on ATP of the light-emitting luciferase-catalyzed luciferin-conversion to oxyluciferin, which can be easily visualized. The assay exhibits a robust EF-dose response decrease in luminescence, which may be specifically reverted by anti-EF antibodies. The application of the assay is exemplified in: (a) determining the presence of EF in B. anthracis cultures, or its absence in cultures of EF-defective strains; (b) evaluating the anti-EF humoral response in experimental animals infected\/vaccinated with B. anthracis; and (c) rapid discrimination between EF producing and non-producing bacterial colonies. Furthermore, the assay may be amenable with high-throughput screening for EF inhibitory molecules.<\/jats:p>","DOI":"10.3390\/toxins8080243","type":"journal-article","created":{"date-parts":[[2016,8,18]],"date-time":"2016-08-18T09:58:54Z","timestamp":1471514334000},"page":"243","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A Simple Luminescent Adenylate-Cyclase Functional Assay for Evaluation of Bacillus anthracis Edema Factor Activity"],"prefix":"10.3390","volume":"8","author":[{"given":"Ma\u2019ayan","family":"Israeli","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shahar","family":"Rotem","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Uri","family":"Elia","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3435-8599","authenticated-orcid":false,"given":"Erez","family":"Bar-Haim","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ofer","family":"Cohen","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Theodor","family":"Chitlaru","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2016,8,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1016\/S1369-5274(00)00168-5","article-title":"Bacillus anthracis, a bug with attitude!","volume":"4","author":"Baillie","year":"2001","journal-title":"Curr. 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