{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T11:17:45Z","timestamp":1788434265975,"version":"build-2803163510"},"reference-count":20,"publisher":"American Chemical Society (ACS)","issue":"17","license":[{"start":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T00:00:00Z","timestamp":1786406400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T00:00:00Z","timestamp":1786406400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T00:00:00Z","timestamp":1786406400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/100000165","name":"Division of Chemistry","doi-asserted-by":"publisher","award":["CHE-1808449"],"award-info":[{"award-number":["CHE-1808449"]}],"id":[{"id":"10.13039\/100000165","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000165","name":"Division of Chemistry","doi-asserted-by":"publisher","award":["CHE-2317422"],"award-info":[{"award-number":["CHE-2317422"]}],"id":[{"id":"10.13039\/100000165","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000740","name":"University of St Andrews","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/501100000740","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Fluorinated succinate analogues were evaluated as mechanistic probes of Mycobacterium tuberculosis isocitrate lyase (MtICL). However, 2,2-difluorosuccinate (1; Ki = 6.1 mM) and 2,2,3-trifluorosuccinate (2; Ki = 23.5 \u03bcM) act as reversible noncompetitive inhibitors and meso-2,3-difluorosuccinate (3) displayed slow-onset reversible inhibition (Ki = 30 \u03bcM), the 2-fluorosuccinate enantiomers ((R)-4 and (S)-4) produced time-dependent irreversible inactivation. Inactivation by 4 was observable under turnover conditions in the presence of glyoxylate and succinate, consistent with a two-step kinetic mechanism. The S enantiomer inactivated more efficiently than (R)-4, consistent with stereoelectronic alignment required for elimination of HF following abstraction of the pro-S proton. 1H NMR analysis detected maleate formation from (S)-4, and mass spectrometry revealed a +132 Da adduct consistent with covalent modification of Cys191. Notably, kinact\/KI values for 4 exceeded that measured for maleate, indicating that covalent capture occurs from an enzyme-bound intermediate prior to product release. These results support a mechanism in which fluorine substitution redirects the enolate-generating half-reaction of MtICL toward elimination and covalent modification. (S)-2-Fluorosuccinate therefore represents a succinate-analogue mechanism-based inactivator that exploits a catalytic step distinct from previously described isocitrate-analogue inhibitors.<\/jats:p>","DOI":"10.1021\/acs.biochem.6c00478","type":"journal-article","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T15:02:10Z","timestamp":1786460530000},"page":"2747-2755","source":"Crossref","is-referenced-by-count":0,"title":["Fluorine-Induced\nElimination Drives Mechanism-Based\nInactivation of Isocitrate Lyase"],"prefix":"10.1021","volume":"65","author":[{"given":"Kolambapatabandige Gayantha Shamin","family":"Fernando","sequence":"first","affiliation":[{"name":"The State University of New York , , , ,","place":["Buffalo, New York, United States, 14260-3000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yohann J. G.","family":"Renault","sequence":"additional","affiliation":[{"name":"University of St Andrews , , North Haugh, , Fife ,","place":["St Andrews, U.K., KY16 9ST"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel","family":"Molino","sequence":"additional","affiliation":[{"name":"The State University of New York , , , ,","place":["Buffalo, New York, United States, 14260-3000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0510-5552","authenticated-orcid":true,"given":"David","family":"O\u2019Hagan","sequence":"additional","affiliation":[{"name":"University of St Andrews , , North Haugh, , Fife ,","place":["St Andrews, U.K., KY16 9ST"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2559-4605","authenticated-orcid":true,"given":"Andrew S.","family":"Murkin","sequence":"additional","affiliation":[{"name":"The State University of New York , , , ,","place":["Buffalo, New York, United States, 14260-3000"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,11]]},"reference":[{"key":"2026090306415870500_cit1","doi-asserted-by":"publisher","first-page":"691","DOI":"10.1038\/nri.2017.69","article-title":"Heterogeneity in Tuberculosis","volume":"17","author":"Cadena","year":"2017","journal-title":"Nat. Rev. Immunol."},{"key":"2026090306415870500_cit2","doi-asserted-by":"publisher","first-page":"735","DOI":"10.1038\/35021074","article-title":"Persistence of Mycobacterium tuberculosis in Macrophages and Mice Requires\nthe Glyoxylate Shunt Enzyme Isocitrate Lyase","volume":"406","author":"McKinney","year":"2000","journal-title":"Nature"},{"key":"2026090306415870500_cit3","doi-asserted-by":"publisher","first-page":"638","DOI":"10.1038\/nm1252","article-title":"Mycobacterium tuberculosis Isocitrate\nLyases 1 and 2 Are Jointly Required for in Vivo Growth and Virulence","volume":"11","author":"Munoz-Elias","year":"2005","journal-title":"Nat. Med."},{"key":"2026090306415870500_cit4","doi-asserted-by":"publisher","first-page":"1042","DOI":"10.1128\/iai.37.3.1042-1049.1982","article-title":"Glyoxylate Metabolism\nand Adaptation of Mycobacterium tuberculosis to Survival under Anaerobic Conditions","volume":"37","author":"Wayne","year":"1982","journal-title":"Infect. Immun."},{"key":"2026090306415870500_cit5","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1038\/77964","article-title":"Structure\nof Isocitrate Lyase, a Persistence Factor of Mycobacterium tuberculosis","volume":"7","author":"Sharma","year":"2000","journal-title":"Nat. Struct. Biol."},{"key":"2026090306415870500_cit6","doi-asserted-by":"publisher","first-page":"178","DOI":"10.1021\/bi401432t","article-title":"Cysteine Is the\nGeneral Base That Serves in Catalysis\nby Isocitrate Lyase and in Mechanism-Based Inhibition by 3-Nitropropionate","volume":"53","author":"Moynihan","year":"2014","journal-title":"Biochemistry"},{"key":"2026090306415870500_cit7","doi-asserted-by":"publisher","first-page":"463","DOI":"10.1021\/acschembio.0c00740","article-title":"Covalent Inactivation\nof Mycobacterium tuberculosis Isocitrate Lyase by Cis-2, 3-Epoxy-Succinic\nAcid","volume":"16","author":"Pham","year":"2021","journal-title":"ACS Chem. Biol."},{"key":"2026090306415870500_cit8","doi-asserted-by":"publisher","first-page":"1470","DOI":"10.1021\/acschembio.8b00225","article-title":"The Nitro Group\nas a Masked Electrophile in Covalent Enzyme Inhibition","volume":"13","author":"Ray","year":"2018","journal-title":"ACS Chem. Biol."},{"key":"2026090306415870500_cit9","doi-asserted-by":"publisher","first-page":"7617","DOI":"10.1073\/pnas.1706134114","article-title":"Mechanism-Based\nInactivator of Isocitrate Lyases 1 and 2 from Mycobacterium tuberculosis","volume":"114","author":"Pham","year":"2017","journal-title":"Proc. Natl. Acad. Sci."},{"key":"2026090306415870500_cit10","doi-asserted-by":"publisher","first-page":"17666","DOI":"10.1021\/jacs.1c07970","article-title":"Mechanism-Based\nInactivation of Mycobacterium tuberculosis Isocitrate\nLyase 1 by (2r,3s)-2-Hydroxy-3-(Nitromethyl)Succinic Acid","volume":"143","author":"Mellott","year":"2021","journal-title":"J. Am. Chem. Soc."},{"key":"2026090306415870500_cit11","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1039\/D0MD00301H","article-title":"Itaconate Is a Covalent Inhibitor of the Mycobacterium\ntuberculosis Isocitrate Lyase","volume":"12","author":"Kwai","year":"2021","journal-title":"RSC Med. Chem."},{"key":"2026090306415870500_cit12","doi-asserted-by":"publisher","first-page":"308","DOI":"10.1039\/B711844A","article-title":"Understanding\nOrganofluorine Chemistry. An Introduction to the C-F Bond","volume":"37","author":"O\u2019Hagan","year":"2008","journal-title":"Chem. Soc. Rev."},{"key":"2026090306415870500_cit13","doi-asserted-by":"publisher","first-page":"6315","DOI":"10.1021\/acs.jmedchem.9b01877","article-title":"Metabolic and Pharmaceutical\nAspects of Fluorinated Compounds","volume":"63","author":"Johnson","year":"2020","journal-title":"J. Med. Chem."},{"key":"2026090306415870500_cit14","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1093\/dnares\/dsi012","article-title":"Complete Set\nof Orf Clones of Escherichia Coli Aska Library (a Complete\nS Et of E. Coli K-12 Orf a Rchive): Unique Resources for Biological\nResearch","volume":"12","author":"Kitagawa","year":"2006","journal-title":"DNA Res."},{"key":"2026090306415870500_cit15","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1186\/1860-5397-2-19","article-title":"The Vicinal Difluoro\nMotif: The Synthesis and Conformation of Erythro- and Threo- Diastereoisomers\nof 1,2-Difluorodiphenylethanes, 2,3-Difluorosuccinic Acids and Their\nDerivatives","volume":"2","author":"O\u2019Hagan","year":"2006","journal-title":"Beilstein J. Org. Chem."},{"key":"2026090306415870500_cit16","doi-asserted-by":"publisher","first-page":"8826","DOI":"10.1021\/ja051805f","article-title":"Enantioselective Organocatalytic Alpha-Fluorination\nof Aldehydes","volume":"127","author":"Beeson","year":"2005","journal-title":"J. Am. Chem. Soc."},{"key":"2026090306415870500_cit17","doi-asserted-by":"publisher","first-page":"1532","DOI":"10.1007\/s00044-023-03103-0","article-title":"Direct Syntheses\nof Stereoisomers of 3-Fluoro Gaba and \u03b2-Fluoroamine Analogues\nof the Calcium Receptor (Car) Agonists, Cinacalcet, Tecalcet, Fendiline\nand Nps R-467","volume":"32","author":"Renault","year":"2023","journal-title":"Med. Chem. Res."},{"key":"2026090306415870500_cit18","doi-asserted-by":"publisher","first-page":"3936","DOI":"10.1021\/jo00332a045","article-title":"Greatly\nImproved Procedure for Ruthenium Tetroxide Catalyzed Oxidations of\nOrganic Compounds","volume":"46","author":"Carlsen","year":"1981","journal-title":"J. Org. Chem."},{"key":"2026090306415870500_cit19","doi-asserted-by":"publisher","first-page":"7538","DOI":"10.1021\/ja00230a045","article-title":"Vicinal Diol Cyclic Sulfates. Like Epoxides Only More\nReactive","volume":"110","author":"Gao","year":"1988","journal-title":"J. Am. Chem. Soc."},{"key":"2026090306415870500_cit20","doi-asserted-by":"publisher","first-page":"1592","DOI":"10.1021\/acs.biochem.6c00181","article-title":"Covalent Inhibition of Mycobacterium tuberculosis Isocitrate Lyase by Maleate Reveals Enolate Reactivity during Catalysis","volume":"65","author":"Fernando","year":"2026","journal-title":"Biochemistry"}],"container-title":["Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.biochem.6c00478","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/65\/17\/2747\/66487825\/acs.biochem.6c00478.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/65\/17\/2747\/66487825\/acs.biochem.6c00478.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T10:42:02Z","timestamp":1788432122000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/bichaw\/article\/65\/17\/2747\/5252136\/Fluorine-Induced-Elimination-Drives-Mechanism"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,11]]},"references-count":20,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,8,11]]},"published-print":{"date-parts":[[2026,9,1]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.biochem.6c00478","relation":{},"ISSN":["0006-2960","1520-4995"],"issn-type":[{"value":"0006-2960","type":"print"},{"value":"1520-4995","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,11]]}}}