{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T14:09:15Z","timestamp":1789135755591,"version":"build-2803163510"},"reference-count":44,"publisher":"American Chemical Society (ACS)","issue":"9","license":[{"start":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T00:00:00Z","timestamp":1787011200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T00:00:00Z","timestamp":1787011200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T00:00:00Z","timestamp":1787011200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["1R15GM148956"],"award-info":[{"award-number":["1R15GM148956"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100011531","name":"University of Alabama","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100011531","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Antimicrobial resistance is an urgent issue in modern-day medicine. One way to address this problem is to repurpose and restore the function of older antibacterials through the specific targeting of antibiotic-modifying enzymes produced by bacteria. Fosfomycin is an antibiotic that is attractive for repurposing because it has been previously approved for use in uncomplicated urinary tract infections (UTIs) and is widely regarded as safe with no known off-target interactions. The primary limitation to fosfomycin usage has been the emergence of fosfomycin resistance enzymes. FosAPa is the fosfomycin resistance enzyme from Pseudomonas aeruginosa. We identified and evaluated several potential inhibitors of FosAPa using high-throughput virtual screening (HTVS) and 31P-NMR. In addition, we obtained X-ray crystal structures of FosAPa with the inhibitors bound in the active site to gain insights into important protein\u2013ligand interactions necessary for the potential structure-based drug design. Preliminary cell-based assays were performed to evaluate the effect that the inhibitors have on the minimum inhibitory concentration (MIC) of fosfomycin in P. aeruginosa. Our results provide an initial scaffold for structure\u2013activity relationships and future compound modifications for combating fosfomycin resistance in P. aeruginosa and other FosA-containing Gram-negative pathogens.<\/jats:p>","DOI":"10.1021\/acsinfecdis.6c00121","type":"journal-article","created":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T19:02:27Z","timestamp":1787079747000},"page":"3043-3052","source":"Crossref","is-referenced-by-count":0,"title":["Inhibition of\nFosfomycin Resistance Protein FosA from\n                    <i>Pseudomonas aeruginosa<\/i>"],"prefix":"10.1021","volume":"12","author":[{"given":"Vanessa","family":"Wiltsie","sequence":"first","affiliation":[{"name":"The University of Alabama , , 250 Hackberry Lane , , ,","place":["Tuscaloosa, Alabama, United States, 35487"]},{"name":"Huntingdon College , , 3104 Huntingdon College Drive , , ,","place":["Montgomery, Alabama, United States, 36106"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatum","family":"Smith","sequence":"additional","affiliation":[{"name":"The University of Alabama , , 250 Hackberry Lane , , ,","place":["Tuscaloosa, Alabama, United States, 35487"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3605-169X","authenticated-orcid":true,"given":"Nishad","family":"Thamban Chandrika","sequence":"additional","affiliation":[{"name":"University of Kentucky , , 789 South Limestone St. , , ,","place":["Lexington, Kentucky, United States, 40536"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6706-811X","authenticated-orcid":true,"given":"Nathaniel C.","family":"Gilbert","sequence":"additional","affiliation":[{"name":"Louisiana State University , , , ,","place":["Baton Rouge, Louisiana, United States, 70803"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7961-5555","authenticated-orcid":true,"given":"Sylvie","family":"Garneau-Tsodikova","sequence":"additional","affiliation":[{"name":"University of Kentucky , , 789 South Limestone St. , , ,","place":["Lexington, Kentucky, United States, 40536"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1964-3896","authenticated-orcid":true,"given":"Matthew K.","family":"Thompson","sequence":"additional","affiliation":[{"name":"The University of Alabama , , 250 Hackberry Lane , , ,","place":["Tuscaloosa, Alabama, United States, 35487"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,18]]},"reference":[{"key":"2026091109093606900_cit1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1128\/CMR.00181-19","article-title":"Antimicrobial\nResistance in ESKAPE Pathogens","volume":"33","author":"De Oliveira","year":"2020","journal-title":"Clin Microbiol\nRev."},{"key":"2026091109093606900_cit2","unstructured":"Centers for Disease Control and Prevention.\n          .  Multidrug Resistant\nPseudomonas aeruginosa; 2019; https:\/\/www.cdc.gov\/antimicrobial-resistance\/media\/pdfs\/pseudomonas-aeruginosa-508.pdf."},{"key":"2026091109093606900_cit3","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1016\/j.biotechadv.2018.11.013","article-title":"Antibiotic\nresistance in Pseudomonas aeruginosa: mechanisms\nand alternative therapeutic strategies","volume":"37","author":"Pang","year":"2019","journal-title":"Biotechnol\nAdv."},{"key":"2026091109093606900_cit4","doi-asserted-by":"publisher","first-page":"582","DOI":"10.1128\/CMR.00040-09","article-title":"Antibacterial-resistant Pseudomonas aeruginosa: clinical impact and complex regulation of chromosomally\nencoded resistance mechanisms","volume":"22","author":"Lister","year":"2009","journal-title":"Clin Microbiol\nRev."},{"key":"2026091109093606900_cit5","doi-asserted-by":"publisher","first-page":"547","DOI":"10.1097\/QCO.0000000000000217","article-title":"Managing Pseudomonas aeruginosa respiratory infections in cystic fibrosis","volume":"28","author":"Langan","year":"2015","journal-title":"Curr. Opin Infect Dis."},{"key":"2026091109093606900_cit6","doi-asserted-by":"publisher","first-page":"fnx124","DOI":"10.1093\/femsle\/fnx124","article-title":"The contribution\nof Pseudomonas aeruginosa virulence factors and host factors in the establishment\nof urinary tract infections","volume":"364","author":"Newman","year":"2017","journal-title":"FEMS Microbiol\nLett."},{"key":"2026091109093606900_cit7","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1128\/CMR.15.2.194-222.2002","article-title":"Lung infections\nassociated with cystic\nfibrosis","volume":"15","author":"Lyczak","year":"2002","journal-title":"Clin Microbiol Rev."},{"key":"2026091109093606900_cit8","doi-asserted-by":"publisher","first-page":"671","DOI":"10.1055\/s-2004-815663","article-title":"Pseudomonas aeruginosa: role in the pathogenesis of the\nCF lung lesion","volume":"24","author":"Currie","year":"2003","journal-title":"Semin Respir Crit Care Med."},{"key":"2026091109093606900_cit9","doi-asserted-by":"publisher","first-page":"300","DOI":"10.3390\/pathogens11030300","article-title":"Pathogenesis\nof the Pseudomonas aeruginosa Biofilm: A Review","volume":"11","author":"Tuon","year":"2022","journal-title":"Pathogens"},{"key":"2026091109093606900_cit10","doi-asserted-by":"publisher","first-page":"1413","DOI":"10.1378\/chest.14-2171","article-title":"Antimicrobial resistance in hospital-acquired\ngram-negative bacterial\ninfections","volume":"147","author":"Mehrad","year":"2015","journal-title":"Chest."},{"key":"2026091109093606900_cit11","doi-asserted-by":"publisher","first-page":"160","DOI":"10.1128\/CMR.00037-09","article-title":"Three decades of\nbeta-lactamase inhibitors","volume":"23","author":"Drawz","year":"2010","journal-title":"Clin Microbiol Rev."},{"key":"2026091109093606900_cit12","doi-asserted-by":"publisher","first-page":"122","DOI":"10.1126\/science.166.3901.122","article-title":"Phosphonomycin, a new\nantibiotic produced by strains\nof streptomyces","volume":"166","author":"Hendlin","year":"1969","journal-title":"Science."},{"key":"2026091109093606900_cit13","doi-asserted-by":"publisher","first-page":"1069","DOI":"10.1086\/527442","article-title":"Fosfomycin:\nuse beyond urinary tract and gastrointestinal infections","volume":"46","author":"Falagas","year":"2008","journal-title":"Clin Infect Dis."},{"key":"2026091109093606900_cit14","doi-asserted-by":"publisher","first-page":"22","DOI":"10.1016\/j.ijantimicag.2018.09.013","article-title":"Resistance to fosfomycin: Mechanisms, Frequency and\nClinical Consequences","volume":"53","author":"Falagas","year":"2019","journal-title":"Int. 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A review\nof its antibacterial activity, pharmacokinetic properties and therapeutic\nefficacy as a single-dose oral treatment for acute uncomplicated lower\nurinary tract infections","volume":"53","author":"Patel","year":"1997","journal-title":"Drugs."},{"key":"2026091109093606900_cit17","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1016\/j.jcf.2011.11.003","article-title":"Antimicrobial activity\nof fosfomycin and tobramycin in combination against cystic fibrosis\npathogens under aerobic and anaerobic conditions","volume":"11","author":"McCaughey","year":"2012","journal-title":"J. Cyst Fibros."},{"key":"2026091109093606900_cit18","doi-asserted-by":"publisher","first-page":"171","DOI":"10.1164\/rccm.201105-0924OC","article-title":"Fosfomycin\/tobramycin for inhalation\nin patients with cystic fibrosis with pseudomonas airway infection","volume":"185","author":"Trapnell","year":"2012","journal-title":"Am. J. Respir Crit Care Med."},{"key":"2026091109093606900_cit19","doi-asserted-by":"publisher","first-page":"1552","DOI":"10.1128\/AAC.32.10.1552","article-title":"Formation of an\nadduct between fosfomycin and glutathione:\na new mechanism of antibiotic resistance in bacteria","volume":"32","author":"Arca","year":"1988","journal-title":"Antimicrob. Agents Chemother."},{"key":"2026091109093606900_cit20","doi-asserted-by":"publisher","first-page":"3042","DOI":"10.1093\/jac\/dkv221","article-title":"In vitro pharmacodynamics of fosfomycin\nagainst clinical\nisolates of Pseudomonas aeruginosa","volume":"70","author":"Walsh","year":"2015","journal-title":"J. Antimicrob. Chemother."},{"key":"2026091109093606900_cit21","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1016\/S1569-1993(02)00143-1","article-title":"Fosfomycin\ntherapy for multiresistant Pseudomonas aeruginosa in cystic fibrosis","volume":"2","author":"Mirakhur","year":"2003","journal-title":"J. Cyst Fibros."},{"key":"2026091109093606900_cit22","doi-asserted-by":"publisher","first-page":"285","DOI":"10.1007\/s10156-007-0534-7","article-title":"Treatment\nof Pseudomonas aeruginosa biofilms with a combination of fluoroquinolones\nand fosfomycin in a rat urinary tract infection model","volume":"13","author":"Mikuniya","year":"2007","journal-title":"J. Infect Chemother."},{"key":"2026091109093606900_cit23","doi-asserted-by":"publisher","first-page":"947","DOI":"10.1039\/D3MD00113J","article-title":"Identification and\nanalysis of small molecule inhibitors of FosB from Staphylococcus aureus","volume":"14","author":"Travis","year":"2023","journal-title":"RSC Med. Chem."},{"key":"2026091109093606900_cit24","doi-asserted-by":"publisher","first-page":"130444","DOI":"10.1016\/j.bbagen.2023.130444","article-title":"Small molecule inhibitors\nof the fosfomycin resistance enzyme FosM from Mycobacterium abscessus","volume":"1867","author":"Chiasson","year":"2023","journal-title":"Biochim Biophys Acta Gen\nSubj."},{"key":"2026091109093606900_cit25","doi-asserted-by":"publisher","first-page":"1739","DOI":"10.1021\/jm0306430","article-title":"Glide:\na new approach for rapid, accurate docking and scoring. 1. Method\nand assessment of docking accuracy","volume":"47","author":"Friesner","year":"2004","journal-title":"J. Med.\nChem."},{"key":"2026091109093606900_cit26","doi-asserted-by":"publisher","first-page":"1750","DOI":"10.1021\/jm030644s","article-title":"Glide: a new approach for rapid,\naccurate docking and scoring. 2. Enrichment factors in database screening","volume":"47","author":"Halgren","year":"2004","journal-title":"J. Med. 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Soc."},{"key":"2026091109093606900_cit29","doi-asserted-by":"publisher","first-page":"1948","DOI":"10.1039\/C9MD00372J","article-title":"Characterization of the genomically encoded fosfomycin\nresistance enzyme from Mycobacterium abscessus","volume":"10","author":"Travis","year":"2019","journal-title":"Medchemcomm."},{"key":"2026091109093606900_cit30","doi-asserted-by":"publisher","first-page":"17786","DOI":"10.1074\/jbc.M501052200","article-title":"Functional analysis of active site residues of the\nfosfomycin resistance enzyme FosA from Pseudomonas aeruginosa","volume":"280","author":"Beharry","year":"2005","journal-title":"J. Biol. Chem."},{"key":"2026091109093606900_cit31","doi-asserted-by":"publisher","first-page":"13625","DOI":"10.1021\/bi001814v","article-title":"Mechanistic\ndiversity in a metalloenzyme superfamily","volume":"39","author":"Armstrong","year":"2000","journal-title":"Biochemistry."},{"key":"2026091109093606900_cit32","doi-asserted-by":"publisher","first-page":"755","DOI":"10.1021\/bi4015852","article-title":"Structure\nand function of the genomically encoded fosfomycin resistance enzyme,\nFosB, from Staphylococcus aureus","volume":"53","author":"Thompson","year":"2014","journal-title":"Biochemistry."},{"key":"2026091109093606900_cit33","doi-asserted-by":"publisher","first-page":"3778","DOI":"10.1021\/jo980014b","article-title":"Regiochemical and Stereochemical\nCourse of the Reaction Catalyzed by the Fosfomycin Resistance Protein,\nFosA","volume":"63","author":"Bernat","year":"1998","journal-title":"Journal of Organic Chemistry."},{"key":"2026091109093606900_cit34","doi-asserted-by":"publisher","first-page":"13666","DOI":"10.1021\/bi048767h","article-title":"Phosphonoformate:\na minimal transition state analogue inhibitor of the fosfomycin resistance\nprotein, FosA","volume":"43","author":"Rigsby","year":"2004","journal-title":"Biochemistry."},{"key":"2026091109093606900_cit35","doi-asserted-by":"publisher","first-page":"583","DOI":"10.1038\/s41586-021-03819-2","article-title":"Highly accurate protein\nstructure prediction with AlphaFold","volume":"596","author":"Jumper","year":"2021","journal-title":"Nature."},{"key":"2026091109093606900_cit36","doi-asserted-by":"publisher","first-page":"816","DOI":"10.1128\/jb.115.3.816-823.1973","article-title":"Three kinds of controls\naffecting the expression of\nthe glp regulon in Escherichia coli","volume":"115","author":"Freedberg","year":"1973","journal-title":"J. Bacteriol."},{"key":"2026091109093606900_cit37","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1128\/AAC.01424-17","article-title":"Inhibition of Fosfomycin Resistance Protein FosA by Phosphonoformate\n(Foscarnet) in Multidrug-Resistant Gram-Negative Pathogens","volume":"61","author":"Ito","year":"2017","journal-title":"Antimicrob. Agents Chemother."},{"key":"2026091109093606900_cit38","doi-asserted-by":"publisher","first-page":"1204","DOI":"10.1107\/S0907444913000061","article-title":"How good are my data and what is the resolution?","volume":"69","author":"Evans","year":"2013","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"2026091109093606900_cit39","doi-asserted-by":"publisher","first-page":"861","DOI":"10.1107\/S2059798319011471","article-title":"Macromolecular structure\ndetermination using X-rays,\nneutrons and electrons: recent developments in Phenix","volume":"75","author":"Liebschner","year":"2019","journal-title":"Acta Crystallogr. D Struct Biol."},{"key":"2026091109093606900_cit40","doi-asserted-by":"publisher","first-page":"2276","DOI":"10.1107\/S0907444913022750","article-title":"Phaser.MRage: automated\nmolecular replacement","volume":"69","author":"Bunkoczi","year":"2013","journal-title":"Acta Crystallogr. D Biol.\nCrystallogr."},{"key":"2026091109093606900_cit41","doi-asserted-by":"publisher","first-page":"486","DOI":"10.1107\/S0907444910007493","article-title":"Features and development\nof Coot","volume":"66","author":"Emsley","year":"2010","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"2026091109093606900_cit42","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1107\/S2059798318006551","article-title":"Real-space refinement in PHENIX for\ncryo-EM and crystallography","volume":"74","author":"Afonine","year":"2018","journal-title":"Acta Crystallogr.\nD Struct Biol."},{"key":"2026091109093606900_cit43","doi-asserted-by":"publisher","first-page":"1605","DOI":"10.1002\/jcc.20084","article-title":"UCSF Chimera-a visualization\nsystem\nfor exploratory research and analysis","volume":"25","author":"Pettersen","year":"2004","journal-title":"J. Comput.\nChem."},{"key":"2026091109093606900_cit44","unstructured":"Wiltsie, V. E.\n          \n          Identification\nand Inhibition of Fosfomycin Resistance Enzymes for Structure-Based\nDrug Design; Ph.D. Dissertation, University of Alabama,\nTuscaloosa, AL, 2025; https:\/\/www.proquest.com\/pqdtglobal\/docview\/3244193948."}],"container-title":["ACS Infectious Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsinfecdis.6c00121","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article-pdf\/12\/9\/3043\/66854198\/acsinfecdis.6c00121.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article-pdf\/12\/9\/3043\/66854198\/acsinfecdis.6c00121.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T13:32:44Z","timestamp":1789133564000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article\/12\/9\/3043\/5275738\/Inhibition-of-Fosfomycin-Resistance-Protein-FosA"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,18]]},"references-count":44,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,8,18]]},"published-print":{"date-parts":[[2026,9,11]]}},"URL":"https:\/\/doi.org\/10.1021\/acsinfecdis.6c00121","relation":{},"ISSN":["2373-8227"],"issn-type":[{"value":"2373-8227","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,18]]}}}