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Arginine metabolism plays a pivotal role in antibiotic susceptibility and biofilm development, making this pathway an attractive therapeutic target. Here, we demonstrate that blestriarene B exerts potent antibacterial activity against S. aureus both in vitro and in vivo. Data-independent acquisition (DIA) proteomics analysis suggests that the antibacterial mechanism of blestriarene B involves an interaction with the HTH-type transcriptional regulator ArcR, which is a key regulator of arginine catabolism. Comparative analyses between wild-type and arcR-deficient S. aureus strains confirmed that ArcR is the molecular target of blestriarene B. Further mechanistic studies revealed that blestriarene B inhibits biofilm formation, impairs cellular ATP production, and ultimately induces bacterial death by binding to ArcR. Collectively, these findings identify blestriarene B as a promising antibacterial drug and validate ArcR as a novel target for anti-S. aureus therapeutics.<\/jats:p>","DOI":"10.1021\/acs.jproteome.6c00236","type":"journal-article","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T20:01:40Z","timestamp":1786478500000},"page":"4757-4771","source":"Crossref","is-referenced-by-count":0,"title":["Targeting\nArginine Metabolism via ArcR: Blestriarene\nB Suppresses\n                    <i>Staphylococcus aureus<\/i>\n                    Biofilm\nFormation and Cellular ATP Production"],"prefix":"10.1021","volume":"25","author":[{"given":"Huan-Huan","family":"Wang","sequence":"first","affiliation":[{"name":"Zhuhai Campus of Zunyi Medical University , , , ,","place":["Zhuhai, Guangdong, China, 519041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Ju","sequence":"additional","affiliation":[{"name":"Zhuhai Campus of Zunyi Medical University , , , ,","place":["Zhuhai, Guangdong, China, 519041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Liu","sequence":"additional","affiliation":[{"name":"Zhuhai Campus of Zunyi Medical University , , , ,","place":["Zhuhai, Guangdong, China, 519041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Jiang","sequence":"additional","affiliation":[{"name":"Zhuhai Campus of Zunyi Medical University , , , ,","place":["Zhuhai, Guangdong, China, 519041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huan","family":"Yao","sequence":"additional","affiliation":[{"name":"Zhuhai Campus of Zunyi Medical University , , , ,","place":["Zhuhai, Guangdong, China, 519041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shulong","family":"Zhao","sequence":"additional","affiliation":[{"name":"Affiliated Hospital of Xuzhou Medical University , , , ,","place":["Xuzhou, Jiangsu, China, 221002"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guozhen","family":"Cui","sequence":"additional","affiliation":[{"name":"Zhuhai Campus of Zunyi Medical University , , , ,","place":["Zhuhai, Guangdong, China, 519041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tian-Peng","family":"Yin","sequence":"additional","affiliation":[{"name":"Zhuhai Campus of Zunyi Medical University , , , ,","place":["Zhuhai, Guangdong, China, 519041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ze","family":"Wang","sequence":"additional","affiliation":[{"name":"Zhuhai Campus of Zunyi Medical University , , , ,","place":["Zhuhai, Guangdong, China, 519041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gao-Fei","family":"Du","sequence":"additional","affiliation":[{"name":"Xuzhou Medical University , , , ,","place":["Xuzhou, Jiangsu, China, 221002"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0957-759X","authenticated-orcid":true,"given":"Xiao-Yan","family":"Yang","sequence":"additional","affiliation":[{"name":"Zhuhai Campus of Zunyi Medical University , , , ,","place":["Zhuhai, Guangdong, China, 519041"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,11]]},"reference":[{"issue":"3","key":"2026090409140319700_cit1","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1128\/CMR.00134-14","article-title":"Staphylococcus aureus infections:\nepidemiology, pathophysiology, clinical manifestations, and management","volume":"28","author":"Tong","year":"2015","journal-title":"Clin. Microbiol. Rev."},{"issue":"12","key":"2026090409140319700_cit2","doi-asserted-by":"publisher","DOI":"10.1093\/ofid\/ofac653","article-title":"All-Cause\nand Infection-Related Mortality in Staphylococcus aureus Bacteremia, a Multicenter Prospective Cohort Study","volume":"9","author":"van der Vaart","year":"2022","journal-title":"Open Forum Infect. 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Neurosci."},{"key":"2026090409140319700_cit18","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2023.125407","article-title":"Extraction,\nstructural-activity relationships, bioactivities,\nand application prospects of Bletilla striata polysaccharides\nas ingredients for functional products: A review","volume":"245","author":"Zhu","year":"2023","journal-title":"Int. J. Biol. Macromol."},{"key":"2026090409140319700_cit19","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2021.659215","article-title":"Bletilla striata Polysaccharide Promotes Diabetic\nWound Healing Through Inhibition of the NLRP3 Inflammasome","volume":"12","author":"Zhao","year":"2021","journal-title":"Front. Pharmacol."},{"key":"2026090409140319700_cit20","doi-asserted-by":"publisher","DOI":"10.1155\/2020\/5391379","article-title":"A Review of Isolation,\nChemical Properties,\nand Bioactivities of Polysaccharides from Bletilla striata","volume":"2020","author":"Ji","year":"2020","journal-title":"Biomed. Res. 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Ethnopharmacol."},{"issue":"12","key":"2026090409140319700_cit23","doi-asserted-by":"publisher","first-page":"3503","DOI":"10.1016\/0031-9422(89)80373-5","article-title":"Biphenanthrenes\nfrom Bletilla striata","volume":"28","author":"Yamaki","year":"1989","journal-title":"Phytochemistry"},{"key":"2026090409140319700_cit24","volume-title":"M100 Performance Standards\nfor Antimicrobial Susceptibility Testing","author":"Weinstein","year":"2021","edition":"31"},{"issue":"10","key":"2026090409140319700_cit25","doi-asserted-by":"publisher","DOI":"10.1002\/adhm.202000265","article-title":"Multifunctional\nGlyco-Nanosheets to Eradicate Drug-Resistant\nBacteria on Wounds","volume":"9","author":"Guo","year":"2020","journal-title":"Adv. Healthcare Mater."},{"issue":"12","key":"2026090409140319700_cit26","doi-asserted-by":"publisher","first-page":"3680","DOI":"10.1073\/pnas.1424924112","article-title":"Vaccine\ncomposition formulated with a novel\nTLR7-dependent adjuvant induces high and broad protection against Staphylococcus aureus","volume":"112","author":"Bagnoli","year":"2015","journal-title":"Proc. Natl.\nAcad. Sci. U.S.A."},{"key":"2026090409140319700_cit27","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2020.103666","article-title":"Proteomic\ninvestigation into the action mechanism of berberine against Streptococcus pyogenes","volume":"215","author":"Du","year":"2020","journal-title":"J. Proteomics"},{"issue":"4","key":"2026090409140319700_cit28","doi-asserted-by":"publisher","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis\nof relative gene expression data using real-time\nquantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"issue":"10","key":"2026090409140319700_cit29","doi-asserted-by":"publisher","first-page":"6076","DOI":"10.1128\/AEM.70.10.6076-6085.2004","article-title":"Novel cassette-based shuttle vector system for gram-positive\nbacteria","volume":"70","author":"Charpentier","year":"2004","journal-title":"Appl. Environ. 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Theory Comput."},{"issue":"18","key":"2026090409140319700_cit38","doi-asserted-by":"publisher","first-page":"5882","DOI":"10.1021\/acs.jpcb.5b00689","article-title":"The general\nAMBER force field (GAFF)\ncan accurately predict thermodynamic and transport properties of many\nionic liquids","volume":"119","author":"Sprenger","year":"2015","journal-title":"J. Phys. Chem. B"},{"issue":"41","key":"2026090409140319700_cit39","doi-asserted-by":"publisher","first-page":"8070","DOI":"10.1021\/jp9717655","article-title":"Application\nof the RESP Methodology in the Parametrization\nof Organic Solvents","volume":"102","author":"Fox","year":"1998","journal-title":"J. Phys. Chem. B"},{"issue":"10","key":"2026090409140319700_cit40","doi-asserted-by":"publisher","DOI":"10.1063\/1.2464095","article-title":"Langevin stabilization\nof molecular-dynamics\nsimulations of polymers by means of quasisymplectic algorithms","volume":"126","author":"Larini","year":"2007","journal-title":"J. Chem. 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Microbiol."},{"issue":"10325","key":"2026090409140319700_cit46","doi-asserted-by":"publisher","first-page":"629","DOI":"10.1016\/S0140-6736(21)02724-0","article-title":"Global burden of bacterial\nantimicrobial resistance in 2019: a systematic analysis","volume":"399","author":"Ikuta","year":"2022","journal-title":"Lancet"},{"issue":"11","key":"2026090409140319700_cit47","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pmed.1002184","article-title":"Will 10 Million People\nDie a Year due to Antimicrobial Resistance by 2050?","volume":"13","author":"de Kraker","year":"2016","journal-title":"PLoS Med."},{"issue":"2","key":"2026090409140319700_cit48","doi-asserted-by":"publisher","first-page":"125","DOI":"10.2174\/1389557518666181017143141","article-title":"Antimicrobial Effects\nof Chemical Compounds Isolated from Traditional Chinese Herbal Medicine\n(TCHM) Against Drug-Resistant Bacteria: A Review Paper","volume":"19","author":"Zhao","year":"2018","journal-title":"Mini-Rev. Med. Chem."},{"issue":"25","key":"2026090409140319700_cit49","doi-asserted-by":"publisher","DOI":"10.1186\/s12866-022-02515-z","article-title":"Celastrol Combats Methicillin-Resistant Staphylococcus aureus by Targeting Delta(1) -Pyrroline-5-Carboxylate Dehydrogenase","volume":"10","author":"Yuan","year":"2023","journal-title":"Adv. Sci."},{"issue":"32","key":"2026090409140319700_cit50","doi-asserted-by":"publisher","first-page":"6375","DOI":"10.1021\/acs.jafc.6b02147","article-title":"iTRAQ-Based\nProteomics Revealed the Bactericidal Mechanism\nof Sodium New Houttuyfonate against Streptococcus pneumoniae","volume":"64","author":"Yang","year":"2016","journal-title":"J. Agric. Food Chem."},{"issue":"6","key":"2026090409140319700_cit51","doi-asserted-by":"publisher","first-page":"2513","DOI":"10.1042\/BST20240710","article-title":"Regulatory dynamics\nof arginine metabolism in Staphylococcus aureus","volume":"52","author":"Reslane","year":"2024","journal-title":"Biochem. Soc. 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