{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T04:25:50Z","timestamp":1775708750799,"version":"3.50.1"},"reference-count":37,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2022,12,1]],"date-time":"2022-12-01T00:00:00Z","timestamp":1669852800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003130","name":"Research Fund Flanders","doi-asserted-by":"publisher","award":["G0A4616N"],"award-info":[{"award-number":["G0A4616N"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Research Fund Flanders","doi-asserted-by":"publisher","award":["G066619N"],"award-info":[{"award-number":["G066619N"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Research Fund Flanders","doi-asserted-by":"publisher","award":["1S68722N"],"award-info":[{"award-number":["1S68722N"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Research Fund Flanders","doi-asserted-by":"publisher","award":["3M14022"],"award-info":[{"award-number":["3M14022"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Research Fund Flanders","doi-asserted-by":"publisher","award":["32213231"],"award-info":[{"award-number":["32213231"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003130","name":"Research Fund Flanders","doi-asserted-by":"publisher","award":["20100225-7"],"award-info":[{"award-number":["20100225-7"]}],"id":[{"id":"10.13039\/501100003130","id-type":"DOI","asserted-by":"publisher"}]},{"name":"KU Leuven Research Fund","award":["G0A4616N"],"award-info":[{"award-number":["G0A4616N"]}]},{"name":"KU Leuven Research Fund","award":["G066619N"],"award-info":[{"award-number":["G066619N"]}]},{"name":"KU Leuven Research Fund","award":["1S68722N"],"award-info":[{"award-number":["1S68722N"]}]},{"name":"KU Leuven Research Fund","award":["3M14022"],"award-info":[{"award-number":["3M14022"]}]},{"name":"KU Leuven Research Fund","award":["32213231"],"award-info":[{"award-number":["32213231"]}]},{"name":"KU Leuven Research Fund","award":["20100225-7"],"award-info":[{"award-number":["20100225-7"]}]},{"name":"Zhengzhou University Research Fund","award":["G0A4616N"],"award-info":[{"award-number":["G0A4616N"]}]},{"name":"Zhengzhou University Research Fund","award":["G066619N"],"award-info":[{"award-number":["G066619N"]}]},{"name":"Zhengzhou University Research Fund","award":["1S68722N"],"award-info":[{"award-number":["1S68722N"]}]},{"name":"Zhengzhou University Research Fund","award":["3M14022"],"award-info":[{"award-number":["3M14022"]}]},{"name":"Zhengzhou University Research Fund","award":["32213231"],"award-info":[{"award-number":["32213231"]}]},{"name":"Zhengzhou University Research Fund","award":["20100225-7"],"award-info":[{"award-number":["20100225-7"]}]},{"name":"Hercules Foundation of the Flemish Government","award":["G0A4616N"],"award-info":[{"award-number":["G0A4616N"]}]},{"name":"Hercules Foundation of the Flemish Government","award":["G066619N"],"award-info":[{"award-number":["G066619N"]}]},{"name":"Hercules Foundation of the Flemish Government","award":["1S68722N"],"award-info":[{"award-number":["1S68722N"]}]},{"name":"Hercules Foundation of the Flemish Government","award":["3M14022"],"award-info":[{"award-number":["3M14022"]}]},{"name":"Hercules Foundation of the Flemish Government","award":["32213231"],"award-info":[{"award-number":["32213231"]}]},{"name":"Hercules Foundation of the Flemish Government","award":["20100225-7"],"award-info":[{"award-number":["20100225-7"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Mycothiol (MSH), the major cellular thiol in Mycobacterium tuberculosis (Mtb), plays an essential role in the resistance of Mtb to various antibiotics and oxidative stresses. MshC catalyzes the ATP-dependent ligation of 1-O-(2-amino-2-deoxy-\u03b1-d-glucopyranosyl)-d-myo-inositol (GlcN-Ins) with l-cysteine (l-Cys) to form l-Cys-GlcN-Ins, the penultimate step in MSH biosynthesis. The inhibition of MshC is lethal to Mtb. In the present study, five new cysteinyl-sulfonamides were synthesized, and their binding affinity with MshC was evaluated using a thermal shift assay. Two of them bind the target with EC50 values of 219 and 231 \u00b5M. Crystal structures of full-length MshC in complex with these two compounds showed that they were bound in the catalytic site of MshC, inducing dramatic conformational changes of the catalytic site compared to the apo form. In particular, the observed closure of the KMSKS loop was not detected in the published cysteinyl-sulfamoyl adenosine-bound structure, the latter likely due to trypsin treatment. Despite the confirmed binding to MshC, the compounds did not suppress Mtb culture growth, which might be explained by the lack of adequate cellular uptake. Taken together, these novel cysteinyl-sulfonamide MshC inhibitors and newly reported full-length apo and ligand-bound MshC structures provide a promising starting point for the further development of novel anti-tubercular drugs targeting MshC.<\/jats:p>","DOI":"10.3390\/ijms232315095","type":"journal-article","created":{"date-parts":[[2022,12,1]],"date-time":"2022-12-01T05:03:18Z","timestamp":1669870998000},"page":"15095","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Structural Basis of Cysteine Ligase MshC Inhibition by Cysteinyl-Sulfonamides"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8224-9250","authenticated-orcid":false,"given":"Luping","family":"Pang","sequence":"first","affiliation":[{"name":"Research Center of Basic Medicine, Academy of Medical Sciences, College of Medicine, Zhengzhou University, Zhengzhou 450001, China"},{"name":"Biocrystallography, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Herestraat 49, P.O. Box 822, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8833-9353","authenticated-orcid":false,"given":"Stijn","family":"Lenders","sequence":"additional","affiliation":[{"name":"Biocrystallography, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Herestraat 49, P.O. Box 822, 3000 Leuven, Belgium"},{"name":"Medicinal Chemistry, Rega Institute for Medical Research, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Herestraat 49, P.O. Box 1030, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Evgenii M.","family":"Osipov","sequence":"additional","affiliation":[{"name":"Biocrystallography, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Herestraat 49, P.O. Box 822, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1360-0852","authenticated-orcid":false,"given":"Stephen D.","family":"Weeks","sequence":"additional","affiliation":[{"name":"Biocrystallography, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Herestraat 49, P.O. Box 822, 3000 Leuven, Belgium"},{"name":"Pledge Therapeutics, Gaston Geenslaan 1, 3001 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9624-5536","authenticated-orcid":false,"given":"Jef","family":"Rozenski","sequence":"additional","affiliation":[{"name":"Medicinal Chemistry, Rega Institute for Medical Research, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Herestraat 49, P.O. Box 1030, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1266-9513","authenticated-orcid":false,"given":"Tatiana","family":"Piller","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Davie","family":"Cappoen","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Parasitology and Hygiene, Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7189-4652","authenticated-orcid":false,"given":"Sergei V.","family":"Strelkov","sequence":"additional","affiliation":[{"name":"Biocrystallography, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Herestraat 49, P.O. Box 822, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3528-8588","authenticated-orcid":false,"given":"Arthur","family":"Van Aerschot","sequence":"additional","affiliation":[{"name":"Medicinal Chemistry, Rega Institute for Medical Research, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Herestraat 49, P.O. Box 1030, 3000 Leuven, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,1]]},"reference":[{"key":"ref_1","unstructured":"(2020, February 25). WHO|Global Tuberculosis Report 2019. Available online: http:\/\/www.who.int\/tb\/publications\/global_report\/en\/."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"190035","DOI":"10.1183\/16000617.0035-2019","article-title":"Regimens to Treat Multidrug-Resistant Tuberculosis: Past, Present and Future Perspectives","volume":"28","author":"Pontali","year":"2019","journal-title":"Eur. Respir. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"953","DOI":"10.1016\/S0140-6736(19)31882-3","article-title":"Management of Drug-Resistant Tuberculosis","volume":"394","author":"Lange","year":"2019","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1016\/j.chembiol.2017.01.005","article-title":"Fumarase Deficiency Causes Protein and Metabolite Succination and Intoxicates Mycobacterium Tuberculosis","volume":"24","author":"Ruecker","year":"2017","journal-title":"Cell Chem. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.1038\/s41598-017-01179-4","article-title":"Monitoring Global Protein Thiol-Oxidation and Protein S-Mycothiolation in Mycobacterium Smegmatis under Hypochlorite Stress","volume":"7","author":"Hillion","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1723","DOI":"10.1046\/j.1365-2958.2003.03416.x","article-title":"Association of Mycothiol with Protection of Mycobacterium Tuberculosis from Toxic Oxidants and Antibiotics","volume":"47","author":"Buchmeier","year":"2003","journal-title":"Mol. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1016\/j.chom.2015.05.008","article-title":"The Oxidative Stress Network of Mycobacterium Tuberculosis Reveals Coordination between Radical Detoxification Systems","volume":"17","author":"Nambi","year":"2015","journal-title":"Cell Host Microbe"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.freeradbiomed.2013.07.036","article-title":"The Thioredoxin Antioxidant System","volume":"66","author":"Lu","year":"2014","journal-title":"Free. Radic. Biol. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"6736","DOI":"10.1128\/JB.185.22.6736-6740.2003","article-title":"Mycothiol Is Essential for Growth of Mycobacterium Tuberculosis Erdman","volume":"185","author":"Sareen","year":"2003","journal-title":"J. Bacteriol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1046\/j.1365-2958.2003.03425.x","article-title":"Genes Required for Mycobacterial Growth Defined by High Density Mutagenesis","volume":"48","author":"Sassetti","year":"2003","journal-title":"Mol. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"6885","DOI":"10.1021\/bi012212u","article-title":"ATP-Dependent L-Cysteine:1D-Myo-Inosityl 2-Amino-2-Deoxy-Alpha-D-Glucopyranoside Ligase, Mycothiol Biosynthesis Enzyme MshC, Is Related to Class I Cysteinyl-tRNA Synthetases","volume":"41","author":"Sareen","year":"2002","journal-title":"Biochemistry"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2778","DOI":"10.1093\/emboj\/21.11.2778","article-title":"Structural Origins of Amino Acid Selection without Editing by Cysteinyl-tRNA Synthetase","volume":"21","author":"Newberry","year":"2002","journal-title":"EMBO J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3956","DOI":"10.1016\/j.bmc.2011.05.028","article-title":"Evaluation of NTF1836 as an Inhibitor of the Mycothiol Biosynthetic Enzyme MshC in Growing and Non-Replicating Mycobacterium Tuberculosis","volume":"19","author":"Newton","year":"2011","journal-title":"Bioorg. Med. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1177\/1087057109335743","article-title":"Dequalinium, a New Inhibitor of Mycobacterium Tuberculosis Mycothiol Ligase Identified by High-Throughput Screening","volume":"14","author":"Baker","year":"2009","journal-title":"J. Biomol. Screen."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1146\/annurev.biochem.69.1.617","article-title":"Aminoacyl-tRNA Synthesis","volume":"69","author":"Ibba","year":"2000","journal-title":"Annu. Rev. Biochem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"8705","DOI":"10.1021\/bi301180x","article-title":"Structural Diversity and Protein Engineering of the Aminoacyl-tRNA Synthetases","volume":"51","author":"Perona","year":"2012","journal-title":"Biochemistry"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5227","DOI":"10.1016\/j.ejmech.2011.08.049","article-title":"Aminoacyl-tRNA Synthetase Inhibitors as Potential Antibiotics","volume":"46","author":"Vondenhoff","year":"2011","journal-title":"Eur. J. Med. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5365","DOI":"10.1074\/jbc.REV118.002956","article-title":"Progress and Challenges in Aminoacyl-tRNA Synthetase-Based Therapeutics","volume":"294","author":"Francklyn","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1535","DOI":"10.1039\/C6MD00274A","article-title":"Natural Products and Their Derivatives as tRNA Synthetase Inhibitors and Antimicrobial Agents","volume":"7","author":"Cochrane","year":"2016","journal-title":"Med. Chem. Commun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1039\/C9MD00139E","article-title":"Recent Development of Leucyl-tRNA Synthetase Inhibitors as Antimicrobial Agents","volume":"10","author":"Zhang","year":"2019","journal-title":"Medchemcomm"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1016\/j.bcp.2018.06.009","article-title":"Aminoacyl-tRNA Synthetases, Therapeutic Targets for Infectious Diseases","volume":"154","author":"Lee","year":"2018","journal-title":"Biochem. Pharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Pang, L., Weeks, S.D., and Van Aerschot, A. (2021). Aminoacyl-tRNA Synthetases as Valuable Targets for Antimicrobial Drug Discovery. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22041750"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"104806","DOI":"10.1016\/j.bioorg.2021.104806","article-title":"Inhibitors of Aminoacyl-tRNA Synthetases as Antimycobacterial Compounds: An up-to-Date Review","volume":"110","author":"Bouz","year":"2021","journal-title":"Bioorg. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1748","DOI":"10.1021\/jm301756m","article-title":"Identification of Bacteria-Selective Threonyl-tRNA Synthetase Substrate Inhibitors by Structure-Based Design","volume":"56","author":"Teng","year":"2013","journal-title":"J. Med. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1021\/acsmedchemlett.7b00374","article-title":"N-Leucinyl Benzenesulfonamides as Structurally Simplified Leucyl-tRNA Synthetase Inhibitors","volume":"9","author":"Charlton","year":"2018","journal-title":"ACS Med. Chem. Lett."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.jpba.2013.01.006","article-title":"Differential Scanning Fluorimetry: Rapid Screening of Formulations That Promote the Stability of Reference Preparations","volume":"77","author":"Malik","year":"2013","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"13326","DOI":"10.1021\/bi801708f","article-title":"The 1.6 \u00c5 Crystal Structure of Mycobacterium Smegmatis MshC: The Penultimate Enzyme in the Mycothiol Biosynthetic Pathway","volume":"47","author":"Tremblay","year":"2008","journal-title":"Biochemistry"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1107\/S0907444909052925","article-title":"PHENIX: A Comprehensive Python-Based System for Macromolecular Structure Solution","volume":"66","author":"Adams","year":"2010","journal-title":"Acta Crystallogr. D"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"113021","DOI":"10.1016\/j.ejmech.2020.113021","article-title":"Synthesis and Structure-Activity Studies of Novel Anhydrohexitol-Based Leucyl-tRNA Synthetase Inhibitors","volume":"211","author":"Pang","year":"2021","journal-title":"Eur. J. Med. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.pep.2006.12.006","article-title":"Ligation Independent Cloning Vectors for Expression of SUMO Fusions","volume":"53","author":"Weeks","year":"2007","journal-title":"Protein Expr. Purif."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1016\/j.ejmech.2018.02.013","article-title":"Family-Wide Analysis of Aminoacyl-Sulfamoyl-3-Deazaadenosine Analogues as Inhibitors of Aminoacyl-tRNA Synthetases","volume":"148","author":"Zhang","year":"2018","journal-title":"Eur. J. Med. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1038\/s42003-022-03825-8","article-title":"Partitioning of the Initial Catalytic Steps of Leucyl-tRNA Synthetase Is Driven by an Active Site Peptide-Plane Flip","volume":"5","author":"Pang","year":"2022","journal-title":"Commun. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1007\/978-1-62703-691-7_2","article-title":"Stable Expression Clones and Auto-Induction for Protein Production in E. Coli","volume":"1091","author":"Studier","year":"2014","journal-title":"Methods Mol. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1107\/S0907444911007773","article-title":"Data Processing and Analysis with the AutoPROC Toolbox","volume":"67","author":"Vonrhein","year":"2011","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1107\/S0021889807021206","article-title":"Phaser Crystallographic Software","volume":"40","author":"McCoy","year":"2007","journal-title":"J. Appl. Crystallogr."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1107\/S0907444910007493","article-title":"Features and Development of Coot","volume":"66","author":"Emsley","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1916","DOI":"10.1016\/j.str.2017.10.009","article-title":"Validation of Structures in the Protein Data Bank","volume":"25","author":"Gore","year":"2017","journal-title":"Structure"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/23\/15095\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:31:59Z","timestamp":1760146319000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/23\/15095"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,1]]},"references-count":37,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["ijms232315095"],"URL":"https:\/\/doi.org\/10.3390\/ijms232315095","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,1]]}}}