{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,10]],"date-time":"2026-06-10T16:01:06Z","timestamp":1781107266837,"version":"3.54.1"},"reference-count":8,"publisher":"National Academy of Sciences","issue":"10","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2002,5,14]]},"abstract":"<jats:p>\n                    A fluorescence resonance energy transfer assay has been developed for monitoring\n                    <jats:italic>Bacillus anthracis<\/jats:italic>\n                    lethal factor (LF) protease activity. A fluorogenic 16-mer peptide based on the known LF protease substrate MEK1 was synthesized and found to be cleaved by the enzyme at the anticipated site. Extension of this work to a fluorogenic 19-mer peptide, derived, in part, from a consensus sequence of known LF protease targets, produced a much better substrate, cleaving approximately 100 times more efficiently. This peptide sequence was modified further on resin to incorporate donor\/quencher pairs to generate substrates for use in fluorescence resonance energy transfer-based appearance assays. All peptides cleaved at similar rates with signal\/background ranging from 9\u201316 at 100% turnover. One of these substrates, denoted (Cou)Consensus(K(QSY-35)GG)-NH\n                    <jats:sub>2<\/jats:sub>\n                    , was selected for additional assay optimization. A plate-based assay requiring only low nanomolar levels of enzyme was developed for screening and inhibitor characterization.\n                  <\/jats:p>","DOI":"10.1073\/pnas.062171599","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:46:49Z","timestamp":1027680409000},"page":"6603-6606","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":84,"title":["A peptide-based fluorescence resonance energy transfer assay for\n                    <i>Bacillus anthracis<\/i>\n                    lethal factor protease"],"prefix":"10.1073","volume":"99","author":[{"given":"Richard T.","family":"Cummings","sequence":"first","affiliation":[{"name":"Department of High Throughput Screening and Automation, Department of Human and Animal Infectious Disease Research, and President, Merck Research Laboratories, Rahway, NJ 07065-0900"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott P.","family":"Salowe","sequence":"additional","affiliation":[{"name":"Department of High Throughput Screening and Automation, Department of Human and Animal Infectious Disease Research, and President, Merck Research Laboratories, Rahway, NJ 07065-0900"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barry R.","family":"Cunningham","sequence":"additional","affiliation":[{"name":"Department of High Throughput Screening and Automation, Department of Human and Animal Infectious Disease Research, and President, Merck Research Laboratories, Rahway, NJ 07065-0900"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Judyann","family":"Wiltsie","sequence":"additional","affiliation":[{"name":"Department of High Throughput Screening and Automation, Department of Human and Animal Infectious Disease Research, and President, Merck Research Laboratories, Rahway, NJ 07065-0900"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Young Whan","family":"Park","sequence":"additional","affiliation":[{"name":"Department of High Throughput Screening and Automation, Department of Human and Animal Infectious Disease Research, and President, Merck Research Laboratories, Rahway, NJ 07065-0900"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa M.","family":"Sonatore","sequence":"additional","affiliation":[{"name":"Department of High Throughput Screening and Automation, Department of Human and Animal Infectious Disease Research, and President, Merck Research Laboratories, Rahway, NJ 07065-0900"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Douglas","family":"Wisniewski","sequence":"additional","affiliation":[{"name":"Department of High Throughput Screening and Automation, Department of Human and Animal Infectious Disease Research, and President, Merck Research Laboratories, Rahway, NJ 07065-0900"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cameron M.","family":"Douglas","sequence":"additional","affiliation":[{"name":"Department of High Throughput Screening and Automation, Department of Human and Animal Infectious Disease Research, and President, Merck Research Laboratories, Rahway, NJ 07065-0900"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey D.","family":"Hermes","sequence":"additional","affiliation":[{"name":"Department of High Throughput Screening and Automation, Department of Human and Animal Infectious Disease Research, and President, Merck Research Laboratories, Rahway, NJ 07065-0900"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edward M.","family":"Scolnick","sequence":"additional","affiliation":[{"name":"Department of High Throughput Screening and Automation, Department of Human and Animal Infectious Disease Research, and President, Merck Research Laboratories, Rahway, NJ 07065-0900"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2002,5,7]]},"reference":[{"key":"e_1_3_2_1_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199909093411107"},{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.micro.55.1.647"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.5.2374-2378.1998"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj3520739"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.280.5364.734"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1998.9040"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/n35101998"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1177\/108705719900400206"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.062171599","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,12]],"date-time":"2022-04-12T15:17:32Z","timestamp":1649776652000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.062171599"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2002,5,7]]},"references-count":8,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2002,5,14]]}},"alternative-id":["10.1073\/pnas.062171599"],"URL":"https:\/\/doi.org\/10.1073\/pnas.062171599","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2002,5,7]]},"assertion":[{"value":"2002-03-25","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication 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