{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T06:27:21Z","timestamp":1790749641051,"version":"4.1.0"},"reference-count":31,"publisher":"American Chemical Society (ACS)","issue":"11","license":[{"start":{"date-parts":[[2020,11,9]],"date-time":"2020-11-09T00:00:00Z","timestamp":1604880000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,11,20]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The alarming growth of antibiotic resistance that is currently ongoing is a serious threat to human health. One of the most promising novel antibiotic targets is MraY (phospho-MurNAc-pentapeptide-transferase), an essential enzyme in bacterial cell wall synthesis. Through recent advances in biochemical research, there is now structural information available for MraY, and for its human homologue GPT (GlcNAc-1-P-transferase), that opens up exciting possibilities for structure-based drug design. The antibiotic compound tunicamycin is a natural product inhibitor of MraY that is also toxic to eukaryotes through its binding to GPT. In this work, we have used tunicamycin and modified versions of tunicamycin as tool compounds to explore the active site of MraY and to gain further insight into what determines inhibitor potency. We have investigated tunicamycin variants where the following motifs have been modified: the length and branching of the tunicamycin fatty acyl chain, the saturation of the fatty acyl chain, the 6\u2033-hydroxyl group of the GlcNAc ring, and the ring structure of the uracil motif. The compounds are analyzed in terms of how potently they bind to MraY, inhibit the activity of the enzyme, and affect the protein thermal stability. Finally, we rationalize these results in the context of the protein structures of MraY and GPT.<\/jats:p>","DOI":"10.1021\/acschembio.0c00423","type":"journal-article","created":{"date-parts":[[2020,11,9]],"date-time":"2020-11-09T10:41:30Z","timestamp":1604918490000},"page":"2885-2895","source":"Crossref","is-referenced-by-count":22,"title":["Exploring the Active Site of the Antibacterial Target\nMraY by Modified Tunicamycins"],"prefix":"10.1021","volume":"15","author":[{"given":"Jenny","family":"Hering","sequence":"first","affiliation":[{"name":"Structure, Biophysics and FBLG, Discovery Sciences, R&D, AstraZeneca , ,","place":["Gothenburg, Sweden"]},{"name":"University of Gothenburg , , ,","place":["Gothenburg, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elin","family":"Dunevall","sequence":"additional","affiliation":[{"name":"University of Gothenburg , , ,","place":["Gothenburg, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arjan","family":"Snijder","sequence":"additional","affiliation":[{"name":"Discovery Biology, Discovery Sciences, R&D, AstraZeneca , ,","place":["Gothenburg, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Per-Olof","family":"Eriksson","sequence":"additional","affiliation":[{"name":"Structure, Biophysics and FBLG, Discovery Sciences, R&D, AstraZeneca , ,","place":["Gothenburg, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4893-9917","authenticated-orcid":true,"given":"Michael A.","family":"Jackson","sequence":"additional","affiliation":[{"name":"U.S. Department of Agriculture, National Center for Agricultural Utilization Research , , ,","place":["Peoria, Illinois, United States, 61604"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Trina M.","family":"Hartman","sequence":"additional","affiliation":[{"name":"U.S. Department of Agriculture, National Center for Agricultural Utilization Research , , ,","place":["Peoria, Illinois, United States, 61604"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ran","family":"Ting","sequence":"additional","affiliation":[{"name":"Bioland Laboratory , , ,","place":["Guangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongming","family":"Chen","sequence":"additional","affiliation":[{"name":"Bioland Laboratory , , ,","place":["Guangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6255-4357","authenticated-orcid":true,"given":"Neil P. J.","family":"Price","sequence":"additional","affiliation":[{"name":"U.S. Department of Agriculture, National Center for Agricultural Utilization Research , , ,","place":["Peoria, Illinois, United States, 61604"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gisela","family":"Bra\u0308nde\u0301n","sequence":"additional","affiliation":[{"name":"University of Gothenburg , , ,","place":["Gothenburg, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8226-8112","authenticated-orcid":true,"given":"Margareta","family":"Ek","sequence":"additional","affiliation":[{"name":"Structure, Biophysics and FBLG, Discovery Sciences, R&D, AstraZeneca , ,","place":["Gothenburg, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,11,9]]},"reference":[{"key":"2026090213594920700_cit1","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1038\/nrd2201","article-title":"Drugs for\nbad bugs: confronting the challenges of antibacterial discovery","volume":"6","author":"Payne","year":"2007","journal-title":"Nat. Rev. Drug Discovery"},{"key":"2026090213594920700_cit2","doi-asserted-by":"publisher","first-page":"529","DOI":"10.1038\/nrd4572","article-title":"ESKAPEing the labyrinth of antibacterial discovery","volume":"14","author":"Tommasi","year":"2015","journal-title":"Nat. Rev. Drug Discovery"},{"key":"2026090213594920700_cit3","doi-asserted-by":"publisher","first-page":"85","DOI":"10.2174\/187152606784112128","article-title":"Phospho-MurNAc-pentapeptide\ntranslocase\n(MraY) as a target for antibacterial agents and antibacterial proteins","volume":"6","author":"Bugg","year":"2006","journal-title":"Infect. Disord.: Drug Targets"},{"key":"2026090213594920700_cit4","doi-asserted-by":"publisher","first-page":"865","DOI":"10.1038\/s41429-019-0227-3","article-title":"Mechanism of action of nucleoside\nantibacterial natural\nproduct antibiotics","volume":"72","author":"Bugg","year":"2019","journal-title":"J. Antibiot."},{"key":"2026090213594920700_cit5","doi-asserted-by":"publisher","first-page":"215","DOI":"10.7164\/antibiotics.24.215","article-title":"Tunicamycin,\na new antibiotic. I.\nIsolation and characterization of tunicamycin","volume":"24","author":"Takatsuki","year":"1971","journal-title":"J. Antibiot."},{"key":"2026090213594920700_cit6","doi-asserted-by":"publisher","first-page":"1221","DOI":"10.2174\/156802605774463042","article-title":"MraY Inhibitors\nas Novel Antibacterial Agents","volume":"5","author":"Dini","year":"2005","journal-title":"Curr. Top. Med.\nChem."},{"key":"2026090213594920700_cit7","doi-asserted-by":"publisher","first-page":"217","DOI":"10.1038\/s41594-018-0031-y","article-title":"GlcNAc-1-P-transferase-tunicamycin\ncomplex structure reveals basis for inhibition of N-glycosylation","volume":"25","author":"Yoo","year":"2018","journal-title":"Nat. Struct. Mol. Biol."},{"key":"2026090213594920700_cit8","doi-asserted-by":"publisher","first-page":"1426","DOI":"10.1016\/j.drudis.2018.05.020","article-title":"Structural\nbasis for\nselective inhibition of antibacterial target MraY, a membrane-bound\nenzyme involved in peptidoglycan synthesis","volume":"23","author":"Hering","year":"2018","journal-title":"Drug Discovery Today"},{"key":"2026090213594920700_cit9","doi-asserted-by":"publisher","first-page":"1012","DOI":"10.1126\/science.1236501","article-title":"Crystal structure\nof MraY, an essential membrane enzyme for bacterial cell wall synthesis","volume":"341","author":"Chung","year":"2013","journal-title":"Science (Washington, DC, U. 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Commun."},{"key":"2026090213594920700_cit13","doi-asserted-by":"publisher","first-page":"1045","DOI":"10.1016\/j.cell.2018.10.037","article-title":"Structures\nof DPAGT1 Explain Glycosylation Disease\nMechanisms and Advance TB Antibiotic Design","volume":"175","author":"Dong","year":"2018","journal-title":"Cell"},{"key":"2026090213594920700_cit14","doi-asserted-by":"crossref","first-page":"3105","DOI":"10.1016\/S0021-9258(19)81080-3","article-title":"Relationship of the structure and biological activity\nof the natural homologues of tunicamycin","volume":"257","author":"Duksin","year":"1982","journal-title":"J.\nBiol. Chem."},{"key":"2026090213594920700_cit15","doi-asserted-by":"publisher","first-page":"544","DOI":"10.1021\/np50028a020","article-title":"Tunicamycins,\nstreptovirudins, and corynetoxins, a special subclass of nucleoside\nantibiotics","volume":"46","author":"Eckardt","year":"1983","journal-title":"J. Nat. Prod."},{"key":"2026090213594920700_cit16","doi-asserted-by":"publisher","first-page":"1122","DOI":"10.1038\/ja.2017.141","article-title":"Selective catalytic\nhydrogenation of the N-acyl and\nuridyl double bonds in the tunicamycin family of protein N-glycosylation\ninhibitors","volume":"70","author":"Price","year":"2017","journal-title":"J. Antibiot."},{"key":"2026090213594920700_cit17","doi-asserted-by":"publisher","first-page":"637","DOI":"10.1038\/ja.2016.49","article-title":"Quinovosamycins:\nnew tunicamycin-type antibiotics in which the alpha, beta-1\u2033,11\u2032-linked\nN-acetylglucosamine residue is replaced by N-acetylquinovosamine","volume":"69","author":"Price","year":"2016","journal-title":"J. Antibiot."},{"key":"2026090213594920700_cit18","doi-asserted-by":"publisher","first-page":"106","DOI":"10.1021\/cb100269f","article-title":"Synthetic lethal compound combinations\nreveal a fundamental connection between wall teichoic acid and peptidoglycan\nbiosyntheses in Staphylococcus aureus","volume":"6","author":"Campbell","year":"2011","journal-title":"ACS Chem.\nBiol."},{"key":"2026090213594920700_cit19","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1126\/scitranslmed.aad7364","article-title":"TarO-specific\ninhibitors of wall teichoic acid biosynthesis restore beta-lactam\nefficacy against methicillin-resistant staphylococci","volume":"8","author":"Lee","year":"2016","journal-title":"Sci. Transl. Med."},{"key":"2026090213594920700_cit20","doi-asserted-by":"publisher","first-page":"1070","DOI":"10.1038\/ja.2017.101","article-title":"Modified\ntunicamycins\nwith reduced eukaryotic toxicity that enhance the antibacterial activity\nof beta-lactams","volume":"70","author":"Price","year":"2017","journal-title":"J. Antibiot."},{"key":"2026090213594920700_cit21","doi-asserted-by":"publisher","first-page":"807","DOI":"10.1038\/s41429-019-0220-x","article-title":"Synergistic enhancement\nof beta-lactam antibiotics\nby modified tunicamycin analogs TunR1 and TunR2","volume":"72","author":"Price","year":"2019","journal-title":"J. Antibiot."},{"key":"2026090213594920700_cit22","doi-asserted-by":"publisher","first-page":"362","DOI":"10.1016\/0304-4165(87)90044-4","article-title":"Minor modifications to the structure of tunicamycin\nlead to loss of the biological activity of the antibiotic","volume":"923","author":"Hashim","year":"1987","journal-title":"Biochim. Biophys. Acta, Gen. Subj."},{"key":"2026090213594920700_cit23","doi-asserted-by":"crossref","first-page":"6572","DOI":"10.1016\/S0021-9258(18)50406-3","article-title":"Biological activities\nof the two major components of\ntunicamycin","volume":"254","author":"Mahoney","year":"1979","journal-title":"J. Biol. Chem."},{"key":"2026090213594920700_cit24","doi-asserted-by":"publisher","first-page":"662","DOI":"10.1177\/1087057112436885","article-title":"A high-throughput,\nhomogeneous, fluorescence resonance energy transfer-based assay for\nphospho-N-acetylmuramoyl-pentapeptide translocase (MraY)","volume":"17","author":"Shapiro","year":"2012","journal-title":"J. Biomol. Screening"},{"key":"2026090213594920700_cit25","doi-asserted-by":"publisher","first-page":"1093","DOI":"10.1007\/s13238-010-0127-6","article-title":"Characterization\nof the tunicamycin\ngene cluster unveiling unique steps involved in its biosynthesis","volume":"1","author":"Chen","year":"2010","journal-title":"Protein Cell"},{"key":"2026090213594920700_cit26","doi-asserted-by":"publisher","first-page":"274","DOI":"10.7164\/antibiotics.28.274","article-title":"Streptovirudins,\nnew antibiotics with antibacterial and antiviral activity. II. Isolation,\nchemical characterization, and biological activity of streptovirudins","volume":"28","author":"Eckardt","year":"1975","journal-title":"J. Antibiot."},{"key":"2026090213594920700_cit27","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1016\/S0021-9673(01)95411-1","article-title":"High-performance liquid chromatographic\ncomparison\nof the tunicaminyluracil-based antibiotics corynetoxin, tunicamycin,\nstreptovirudin and MM 19290","volume":"268","author":"Cockrum","year":"1983","journal-title":"J. Chromatogr."},{"key":"2026090213594920700_cit28","doi-asserted-by":"publisher","first-page":"2968","DOI":"10.1021\/bi00513a039","article-title":"Biological activities\nof isolated tunicamycin and streptovirudin fractions","volume":"20","author":"Keenan","year":"1981","journal-title":"Biochemistry"},{"key":"2026090213594920700_cit29","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1007\/s12551-020-00619-2","article-title":"Theory and applications of differential\nscanning fluorimetry in early-stage drug discovery","volume":"12","author":"Gao","year":"2020","journal-title":"Biophys. Rev."},{"key":"2026090213594920700_cit30","doi-asserted-by":"publisher","first-page":"22","DOI":"10.3390\/molecules25010022","article-title":"Muraymycin Nucleoside Antibiotics: Structure-Activity Relationship\nfor Variations in the Nucleoside Unit","volume":"25","author":"Heib","year":"2020","journal-title":"Molecules"},{"key":"2026090213594920700_cit31","doi-asserted-by":"publisher","first-page":"1124","DOI":"10.1002\/pro.2297","article-title":"Fluorescent probe\nfor high-throughput screening of\nmembrane protein expression","volume":"22","author":"Backmark","year":"2013","journal-title":"Protein Sci."}],"container-title":["ACS Chemical Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acschembio.0c00423","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/15\/11\/2885\/66949767\/acschembio.0c00423.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/15\/11\/2885\/66949767\/acschembio.0c00423.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T17:59:54Z","timestamp":1788371994000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acbcct\/article\/15\/11\/2885\/589635\/Exploring-the-Active-Site-of-the-Antibacterial"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,9]]},"references-count":31,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2020,11,9]]},"published-print":{"date-parts":[[2020,11,20]]}},"URL":"https:\/\/doi.org\/10.1021\/acschembio.0c00423","relation":{},"ISSN":["1554-8929","1554-8937"],"issn-type":[{"value":"1554-8929","type":"print"},{"value":"1554-8937","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,11,9]]}}}