{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T18:50:31Z","timestamp":1792003831434,"version":"4.4.0"},"reference-count":37,"publisher":"American Chemical Society (ACS)","issue":"40","license":[{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000054","name":"National Cancer Institute","doi-asserted-by":"publisher","award":["P30 CA008748"],"award-info":[{"award-number":["P30 CA008748"]}],"id":[{"id":"10.13039\/100000054","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01 AI196228"],"award-info":[{"award-number":["R01 AI196228"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Tuberculosis, the disease caused by Mycobacterium tuberculosis infection, is a leading cause of death worldwide. While first-line treatments are effective for many patients, multidrug-resistant tuberculosis poses a continued threat to global public health, and new approaches are needed to provide effective chemotherapy against drug-resistant disease. Diisonitrile lipopeptide chalkophores are a novel class of natural products that play critical roles in copper homeostasis and virulence in M. tuberculosis by supplying copper to the heme:Cu oxidase of the respiratory chain. However, the molecular mechanisms by which the bacteria use diisonitriles to mediate copper homeostasis are poorly understood. Herein, we investigate structure\u2013activity relationships of synthetic diisonitriles to identify structural features required for biological function in mycobacterial copper acquisition. We have developed a modular synthetic approach that allows efficient construction of diisonitrile natural products and diverse analogues. We confirmed that a short-chain analogue of the natural products binds tightly to Cu(I) and Cu(II), but not Zn(II), and estimated its Cu(I) binding affinity. We then used these analogues to probe the impacts of diisonitrile acyl chain length, amino acid backbone, and C-terminal motif on the rescue of M. tuberculosis and Mycobacterium marinum growth in copper-restricted conditions. The observed structural preferences align with current knowledge of the corresponding natural product structures in these bacteria, but both species also exhibited surprising flexibility in accepting diverse diisonitrile analogues for this function. This work provides new insights into the structural features of diisonitrile chalkophores required for function in mycobacterial copper homeostasis, and the distinct structural preferences between species, while suggesting some structural plasticity in their cellular copper acquisition systems.<\/jats:p>","DOI":"10.1021\/jacs.6c09506","type":"journal-article","created":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T13:08:13Z","timestamp":1790860093000},"page":"43041-43049","source":"Crossref","is-referenced-by-count":0,"title":["Diisonitrile Chalkophore\nStructure and Function in\nMycobacterial Copper Acquisition"],"prefix":"10.1021","volume":"148","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-5277-1789","authenticated-orcid":true,"given":"Pooja B.","family":"Pandya","sequence":"first","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center , , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]},{"name":"Memorial Sloan Kettering Cancer Center , , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chad E.","family":"Hatch","sequence":"additional","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center , , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John A.","family":"Buglino","sequence":"additional","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center , , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaprak","family":"Ozakman","sequence":"additional","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center , , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-7846-9770","authenticated-orcid":true,"given":"Kate","family":"Ryan","sequence":"additional","affiliation":[{"name":"Gerstner Sloan Kettering Graduate School, Memorial Sloan Kettering Cancer Center , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruofei Faye","family":"Li","sequence":"additional","affiliation":[{"name":"Mechanistic Biology Summer Program , , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0374-5668","authenticated-orcid":true,"given":"Farhan A.","family":"Chowdhury","sequence":"additional","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center , , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael S.","family":"Glickman","sequence":"additional","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center , , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]},{"name":"Gerstner Sloan Kettering Graduate School, Memorial Sloan Kettering Cancer Center , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7956-9659","authenticated-orcid":true,"given":"Derek S.","family":"Tan","sequence":"additional","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center , , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]},{"name":"Memorial Sloan Kettering Cancer Center , , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]},{"name":"Gerstner Sloan Kettering Graduate School, Memorial Sloan Kettering Cancer Center , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]},{"name":"Tri-Institutional Research Program, Memorial Sloan Kettering Cancer Center , 1275 York Avenue , , ,","place":["New York, New York, United States, 10065"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,10,1]]},"reference":[{"key":"2026101414085903600_cit1","unstructured":"World Health Organization.\n          \n          Global Tuberculosis Report 2025; World Health Organization, Geneva, 2025."},{"key":"2026101414085903600_cit2","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1126\/science.1106753","article-title":"A diarylquinoline drug\nactive on the ATP synthase of Mycobacterium tuberculosis","volume":"307","author":"Andries","year":"2005","journal-title":"Science"},{"key":"2026101414085903600_cit3","doi-asserted-by":"publisher","first-page":"893","DOI":"10.1056\/NEJMoa1901814","article-title":"Treatment of highly drug-resistant pulmonary tuberculosis","volume":"382","author":"Conradie","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"2026101414085903600_cit4","doi-asserted-by":"publisher","first-page":"685","DOI":"10.1038\/s41579-022-00731-y","article-title":"Anti-tuberculosis\ntreatment strategies and drug development:\nchallenges and priorities","volume":"20","author":"Dartois","year":"2022","journal-title":"Nat. Rev. Microbiol."},{"key":"2026101414085903600_cit5","doi-asserted-by":"publisher","first-page":"104","DOI":"10.1016\/j.toxlet.2020.01.018","article-title":"Metabolomics\ndescribes previously\nunknown toxicity mechanisms of isoniazid and rifampicin","volume":"322","author":"Combrink","year":"2020","journal-title":"Toxicol. Lett."},{"key":"2026101414085903600_cit6","doi-asserted-by":"publisher","first-page":"RP105794","DOI":"10.7554\/elife.105794","article-title":"Chalkophore-mediated\nrespiratory oxidase flexibility\ncontrols M. tuberculosis virulence","volume":"14","author":"Buglino","year":"2025","journal-title":"eLife"},{"key":"2026101414085903600_cit7","doi-asserted-by":"publisher","first-page":"657","DOI":"10.1038\/s41579-022-00745-6","article-title":"Nutritional immunity: the battle\nfor nutrient metals\nat the host\u2013pathogen interface","volume":"20","author":"Murdoch","year":"2022","journal-title":"Nat.\nRev. Microbiol."},{"key":"2026101414085903600_cit8","doi-asserted-by":"publisher","first-page":"e2022136118","DOI":"10.1073\/pnas.2022136118","article-title":"Mycobacterium tuberculosis VapC4 toxin engages small ORFs to initiate an\nintegrated oxidative\nand copper stress response","volume":"118","author":"Barth","year":"2021","journal-title":"Proc. Natl. Acad.\nSci. U.S.A."},{"key":"2026101414085903600_cit9","doi-asserted-by":"publisher","first-page":"6","DOI":"10.1186\/s40659-020-00274-7","article-title":"CtpB is\na plasma\nmembrane copper (I) transporting P-type ATPase of Mycobacterium tuberculosis","volume":"53","author":"Leo\u0301n-Torres","year":"2020","journal-title":"Biol. Res."},{"key":"2026101414085903600_cit10","doi-asserted-by":"publisher","first-page":"e02513-22","DOI":"10.1128\/mbio.02513-22","article-title":"Diisonitrile lipopeptides mediate\nresistance to copper starvation in pathogenic mycobacteria","volume":"13","author":"Buglino","year":"2022","journal-title":"mBio"},{"key":"2026101414085903600_cit11","doi-asserted-by":"publisher","first-page":"929","DOI":"10.1039\/C4MT00305E","article-title":"Copper homeostasis in Mycobacterium tuberculosis","volume":"7","author":"Shi","year":"2015","journal-title":"Metallomics"},{"key":"2026101414085903600_cit12","doi-asserted-by":"publisher","first-page":"e2110293119","DOI":"10.1073\/pnas.2110293119","article-title":"Kupyaphores are zinc homeostatic\nmetallophores required for colonization of Mycobacterium tuberculosis","volume":"119","author":"Mehdiratta","year":"2022","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026101414085903600_cit13","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1007\/s00775-025-02099-9","article-title":"Copper-chelating natural products","volume":"30","author":"Manley","year":"2025","journal-title":"J. Biol. Inorg. Chem."},{"key":"2026101414085903600_cit14","doi-asserted-by":"publisher","first-page":"e65351","DOI":"10.7554\/elife.65351","article-title":"Integrated\nsensing of host stresses by inhibition of a cytoplasmic two-component\nsystem controls M. tuberculosis acute lung infection","volume":"10","author":"Buglino","year":"2021","journal-title":"eLife"},{"key":"2026101414085903600_cit15","doi-asserted-by":"publisher","first-page":"4606","DOI":"10.1074\/jbc.TM117.000185","article-title":"Methanobactins: Maintaining copper\nhomeostasis in methanotrophs\nand beyond","volume":"293","author":"Kenney","year":"2018","journal-title":"J. Biol. Chem."},{"key":"2026101414085903600_cit16","doi-asserted-by":"publisher","first-page":"7025","DOI":"10.1073\/pnas.1705016114","article-title":"Biosynthesis of isonitrile\nlipopeptides\nby conserved nonribosomal peptide synthetase gene clusters in Actinobacteria","volume":"114","author":"Harris","year":"2017","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026101414085903600_cit17","doi-asserted-by":"publisher","first-page":"5681","DOI":"10.1073\/pnas.1118680109","article-title":"Nonprocessive [2 + 2]e- off-loading reductase\ndomains from mycobacterial nonribosomal peptide synthetases","volume":"109","author":"Chhabra","year":"2012","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026101414085903600_cit18","doi-asserted-by":"publisher","first-page":"1492","DOI":"10.1021\/acschembio.5c00078","article-title":"Enzymatic pathway for kupyaphore\ndegradation in Mycobacterium tuberculosis: Mechanism\nof metal homeostasis and turnover","volume":"20","author":"Bhosale","year":"2025","journal-title":"ACS Chem.\nBiol."},{"key":"2026101414085903600_cit19","doi-asserted-by":"publisher","first-page":"40652","DOI":"10.1021\/jacs.5c13262","article-title":"Kupyaphores\u2013self-assembling\ndiisocyanolipopeptide ZnII ionophores in Mycobacterium tuberculosis ZnII\/CuI\/II homeostasis\nand antibacterial effects","volume":"147","author":"Wong","year":"2025","journal-title":"J. Am. Chem. Soc."},{"key":"2026101414085903600_cit20","doi-asserted-by":"publisher","first-page":"3067","DOI":"10.1021\/acschembio.7b00897","article-title":"Diisonitrile\nnatural product SF2768 functions as a\nchalkophore that mediates copper acquisition in Streptomyces thioluteus","volume":"12","author":"Wang","year":"2017","journal-title":"ACS Chem. Biol."},{"key":"2026101414085903600_cit21","doi-asserted-by":"publisher","first-page":"8731","DOI":"10.1021\/acs.orglett.9b03348","article-title":"Total synthesis\nof the bacterial diisonitrile chalkophore\nSF2768","volume":"21","author":"Xu","year":"2019","journal-title":"Org. Lett."},{"key":"2026101414085903600_cit22","doi-asserted-by":"publisher","first-page":"4975","DOI":"10.1021\/acscatal.4c00645","article-title":"Variation in biosynthesis and metal-binding\nproperties of isonitrile-containing peptides produced by Mycobacteria\nversus Streptomyces","volume":"14","author":"Chen","year":"2024","journal-title":"ACS Catal."},{"key":"2026101414085903600_cit23","doi-asserted-by":"publisher","first-page":"2490","DOI":"10.1021\/acs.jnatprod.5c00904","article-title":"Discovery of isonitrile lipopeptide\nchalkophores from pathogenic\nMycobacteria","volume":"88","author":"Jia","year":"2025","journal-title":"J. Nat. Prod."},{"key":"2026101414085903600_cit24","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1039\/c6mt00208k","article-title":"Methanobactins:\nfrom genome to function","volume":"9","author":"Dassama","year":"2017","journal-title":"Metallomics"},{"key":"2026101414085903600_cit25","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1038\/s41579-024-01090-6","article-title":"Bacterial\nsiderophores: diversity, uptake pathways and applications","volume":"23","author":"Schalk","year":"2025","journal-title":"Nat. Rev. Microbiol."},{"key":"2026101414085903600_cit26","first-page":"10","article-title":"A new antibiotic SF2369 produced\nby Actinomadura","volume":"26","author":"Sasaki","year":"1987","journal-title":"Sci. Rep. Meiji Seika Kaisha"},{"key":"2026101414085903600_cit27","first-page":"1","article-title":"SF2768, a\nnew isonitrile antibiotic obtained from Streptomyces","volume":"34","author":"Tabata","year":"1995","journal-title":"Sci. Rep. Meiji Seika Kaisha"},{"key":"2026101414085903600_cit28","doi-asserted-by":"publisher","first-page":"1554","DOI":"10.1016\/j.tetasy.2007.06.008","article-title":"Parallel synthesis\nof homochiral \u03b2-amino acids","volume":"18","author":"Davies","year":"2007","journal-title":"Tetrahedron:Asymmetry"},{"key":"2026101414085903600_cit29","doi-asserted-by":"publisher","first-page":"7819","DOI":"10.1021\/jo025957u","article-title":"Asymmetric synthesis\nof \u03b2-amino acid derivatives\nincorporating a broad range of substitution patterns by enolate additions\nto tert-butanesulfinyl imines","volume":"67","author":"Tang","year":"2002","journal-title":"J. Org. Chem."},{"key":"2026101414085903600_cit30","doi-asserted-by":"publisher","first-page":"L53","DOI":"10.1016\/S0020-1693(00)84740-4","article-title":"Bidentate isocyanides\n1. Synthesis of disocyanocyclohexanes and some of their complexes","volume":"53","author":"Ohtani","year":"1981","journal-title":"Inorg. Chim. Acta"},{"key":"2026101414085903600_cit31","doi-asserted-by":"publisher","first-page":"2069","DOI":"10.1073\/pnas.0507924103","article-title":"A genetic locus\nrequired for iron acquisition in Mycobacterium tuberculosis","volume":"103","author":"Krithika","year":"2006","journal-title":"Proc.\nNatl. Acad. Sci. U.S.A."},{"key":"2026101414085903600_cit32","doi-asserted-by":"publisher","first-page":"3081","DOI":"10.1021\/ja0390350","article-title":"C-terminal\ndomain of the membrane copper transporter Ctr1 from Saccharomyces cerevisiae binds four Cu(I) Ions as a cuprous-thiolate polynuclear\ncluster: Sub-femtomolar Cu(I) affinity of three proteins involved\nin copper trafficking","volume":"126","author":"Xiao","year":"2004","journal-title":"J. Am. Chem. Soc."},{"key":"2026101414085903600_cit33","doi-asserted-by":"publisher","first-page":"407","DOI":"10.1021\/ic50204a025","article-title":"Electron-transfer reactions of copper(I)\nand copper(III) complexes","volume":"19","author":"Lappin","year":"1980","journal-title":"Inorg. Chem."},{"key":"2026101414085903600_cit34","doi-asserted-by":"publisher","first-page":"1491","DOI":"10.4049\/jimmunol.174.3.1491","article-title":"Elemental analysis of Mycobacterium avium-, Mycobacterium tuberculosis-, and Mycobacterium smegmatis-containing phagosomes indicates pathogen-induced\nmicroenvironments within the host cell\u2019s endosomal system","volume":"174","author":"Wagner","year":"2005","journal-title":"J. Immunol."},{"key":"2026101414085903600_cit35","doi-asserted-by":"publisher","first-page":"3562","DOI":"10.1021\/acs.orglett.8b01341","article-title":"Tandem hydration\nof diisonitriles triggered by isonitrile hydratase in Streptomyces thioluteus","volume":"20","author":"Zhu","year":"2018","journal-title":"Org. Lett."},{"key":"2026101414085903600_cit36","doi-asserted-by":"publisher","first-page":"18473","DOI":"10.1074\/jbc.M113.466912","article-title":"Structures\nof Mycobacterium tuberculosis FadD10 protein reveal\na new type of adenylate-forming enzyme","volume":"288","author":"Liu","year":"2013","journal-title":"J. Biol.\nChem."},{"key":"2026101414085903600_cit37","doi-asserted-by":"publisher","first-page":"213727","DOI":"10.1016\/j.ccr.2020.213727","article-title":"Extracellular Cu2+ pools and their detection:\nFrom current knowledge to next-generation probes","volume":"433","author":"Falcone","year":"2021","journal-title":"Coord. Chem. Rev."}],"container-title":["Journal of the American\nChemical Society"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/jacs.6c09506","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jacsat\/article-pdf\/148\/40\/43041\/68724987\/jacs.6c09506.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jacsat\/article-pdf\/148\/40\/43041\/68724987\/jacs.6c09506.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T18:30:21Z","timestamp":1792002621000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jacsat\/article\/148\/40\/43041\/5443271\/Diisonitrile-Chalkophore-Structure-and-Function-in"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,1]]},"references-count":37,"journal-issue":{"issue":"40","published-online":{"date-parts":[[2026,10,1]]},"published-print":{"date-parts":[[2026,10,14]]}},"URL":"https:\/\/doi.org\/10.1021\/jacs.6c09506","relation":{},"ISSN":["0002-7863","1520-5126"],"issn-type":[{"value":"0002-7863","type":"print"},{"value":"1520-5126","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,1]]}}}