{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T05:25:22Z","timestamp":1791523522309,"version":"4.3.4"},"reference-count":34,"publisher":"Oxford University Press (OUP)","issue":"5","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2015,5,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Objectives<\/jats:title>\n                    <jats:p>The spread of NDM-1 amongst Enterobacteriaceae has highlighted a significant threat to the clinical management of serious infections. The combination of aztreonam and avibactam, a non-\u03b2-lactam \u03b2-lactamase inhibitor, may provide a much-needed therapeutic alternative. This combination was potent against most NDM-containing Enterobacteriaceae, although activity was diminished against many Escherichia coli isolates. These E. coli isolates were characterized to elucidate the mechanism of decreased susceptibility to aztreonam\/avibactam.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>MIC determinations were performed using broth microdilution, and whole-genome sequencing was performed to enable sequence-based analyses.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The decreased susceptibility was not due to avibactam being unable to inhibit the serine \u03b2-lactamases found in the E. coli isolates. Rather, it was manifested by a four-amino-acid insertion in PBP3. This same insertion was also found in non-NDM-containing E. coli that had reduced susceptibility to aztreonam\/avibactam. Construction of an isogenic mutant confirmed that this insertion resulted in decreased susceptibility to aztreonam and several cephalosporins, but had no impact on carbapenem potency. Structural analysis suggests that this insertion will impact the accessibility of the \u03b2-lactam drugs to the transpeptidase pocket of PBP3.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>The acquisition of \u03b2-lactamases is the predominant mechanism of \u03b2-lactam resistance in Enterobacteriaceae. We have demonstrated that small PBP3 changes will affect the susceptibility to a broad range of \u03b2-lactams. These changes were identified in multiple MLST lineages of E. coli, and were enriched in NDM-containing isolates. However, they were not present in other key species of Enterobacteriaceae despite significant conservation among the PBP3 proteins.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/jac\/dku568","type":"journal-article","created":{"date-parts":[[2015,1,30]],"date-time":"2015-01-30T00:19:53Z","timestamp":1422577193000},"page":"1420-1428","source":"Crossref","is-referenced-by-count":188,"title":["Characterization of\n                    <i>Escherichia coli<\/i>\n                    NDM isolates with decreased susceptibility to aztreonam\/avibactam: role of a novel insertion in PBP3"],"prefix":"10.1093","volume":"70","author":[{"given":"Richard A.","family":"Alm","sequence":"first","affiliation":[{"name":"Infection Innovative Medicines Unit, AstraZeneca R&D Boston, Waltham, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michele R.","family":"Johnstone","sequence":"additional","affiliation":[{"name":"Infection Innovative Medicines Unit, AstraZeneca R&D Boston, Waltham, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sushmita D.","family":"Lahiri","sequence":"additional","affiliation":[{"name":"Infection Innovative Medicines Unit, AstraZeneca R&D Boston, Waltham, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2015,1,28]]},"reference":[{"key":"2022020121021980800_DKU568C1","doi-asserted-by":"crossref","first-page":"12478","DOI":"10.3390\/molecules171112478","article-title":"Development of new drugs for an old target: the penicillin binding proteins","volume":"17","author":"Zervosen","year":"2012","journal-title":"Molecules"},{"key":"2022020121021980800_DKU568C2","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1111\/j.1574-6976.2008.00105.x","article-title":"The penicillin-binding proteins: structure and role in peptidoglycan biosynthesis","volume":"32","author":"Sauvage","year":"2008","journal-title":"FEMS Microbiol Rev"},{"key":"2022020121021980800_DKU568C3","doi-asserted-by":"crossref","first-page":"2394","DOI":"10.1093\/jac\/dku158","article-title":"Frequent carriage of resistance mechanisms to \u03b2-lactams and biofilm formation in Haemophilus influenzae causing treatment failure and recurrent otitis media in young children","volume":"69","author":"Garcia-Cobos","year":"2014","journal-title":"J Antimicrob Chemother"},{"key":"2022020121021980800_DKU568C4","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1093\/jac\/dks525","article-title":"Novel mechanisms of resistance to \u03b2-lactam antibiotics in Haemophilus parainfluenzae: \u03b2-lactamase-negative ampicillin resistance and inhibitor-resistant TEM \u03b2-lactamases","volume":"68","author":"Garcia-Cobos","year":"2013","journal-title":"J Antimicrob Chemother"},{"key":"2022020121021980800_DKU568C5","doi-asserted-by":"crossref","first-page":"1202","DOI":"10.1074\/jbc.M805761200","article-title":"Crystal structures of penicillin-binding protein 2 from penicillin-susceptible and -resistant strains of Neisseria gonorrhoeae reveal an unexpectedly subtle mechanism for antibiotic resistance","volume":"284","author":"Powell","year":"2009","journal-title":"J Biol Chem"},{"key":"2022020121021980800_DKU568C6","doi-asserted-by":"crossref","first-page":"899","DOI":"10.1016\/j.jmb.2009.02.024","article-title":"PBP active site flexibility as the key mechanism for \u03b2-lactam resistance in pneumococci","volume":"387","author":"Contreras-Martel","year":"2009","journal-title":"J Mol Biol"},{"key":"2022020121021980800_DKU568C7","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1111\/j.1469-0691.2006.01573.x","article-title":"The molecular evolution of methicillin-resistant Staphylococcus aureus","volume":"13","author":"Deurenberg","year":"2007","journal-title":"Clin Microbiol Infect"},{"key":"2022020121021980800_DKU568C8","doi-asserted-by":"crossref","first-page":"e1000855","DOI":"10.1371\/journal.ppat.1000855","article-title":"A timescale for evolution, population expansion, and spatial spread of an emerging clone of methicillin-resistant Staphylococcus aureus","volume":"6","author":"Nubel","year":"2010","journal-title":"PLoS Pathog"},{"key":"2022020121021980800_DKU568C9","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1128\/CMR.00036-08","article-title":"AmpC \u03b2-lactamases","volume":"22","author":"Jacoby","year":"2009","journal-title":"Clin Microbiol Rev"},{"key":"2022020121021980800_DKU568C10","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1016\/j.mib.2010.09.006","article-title":"Alarming \u03b2-lactamase-mediated resistance in multidrug-resistant Enterobacteriaceae","volume":"13","author":"Bush","year":"2010","journal-title":"Curr Opin Microbiol"},{"key":"2022020121021980800_DKU568C11","doi-asserted-by":"crossref","first-page":"1774","DOI":"10.1128\/AAC.02123-13","article-title":"In vitro activities of ceftazidime-avibactam and aztreonam-avibactam against 372 Gram-negative bacilli collected in 2011 and 2012 from 11 teaching hospitals in China","volume":"58","author":"Wang","year":"2014","journal-title":"Antimicrob Agents Chemother"},{"key":"2022020121021980800_DKU568C12","doi-asserted-by":"crossref","first-page":"5046","DOI":"10.1128\/AAC.00774-09","article-title":"Characterization of a new metallo-\u03b2-lactamase gene, blaNDM-1, and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India","volume":"53","author":"Yong","year":"2009","journal-title":"Antimicrob Agents Chemother"},{"key":"2022020121021980800_DKU568C13","doi-asserted-by":"crossref","DOI":"10.2807\/1560-7917.ES2014.19.20.20809","article-title":"New Delhi metallo-\u03b2-lactamase around the world: an eReview using Google Maps","volume":"19","author":"Berrazeg","year":"2014","journal-title":"Euro Surveill"},{"key":"2022020121021980800_DKU568C14","doi-asserted-by":"crossref","first-page":"e325","DOI":"10.1016\/j.ijid.2012.11.025","article-title":"Emergence of New Delhi metallo-\u03b2-lactamase type 1-producing Enterobacteriaceae and non-Enterobacteriaceae: Gglobal case detection and bacterial surveillance","volume":"17","author":"Bushnell","year":"2013","journal-title":"Int J Infect Dis"},{"key":"2022020121021980800_DKU568C15","doi-asserted-by":"crossref","first-page":"870","DOI":"10.3201\/eid1906.121515","article-title":"New Delhi metallo-\u03b2-lactamase-producing Enterobacteriaceae, United States","volume":"19","author":"Rasheed","year":"2013","journal-title":"Emerg Infect Dis"},{"key":"2022020121021980800_DKU568C16","doi-asserted-by":"crossref","first-page":"2621","DOI":"10.1128\/AAC.00029-07","article-title":"Binding of ceftobiprole and comparators to the penicillin-binding proteins of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pneumoniae","volume":"51","author":"Davies","year":"2007","journal-title":"Antimicrob Agents Chemother"},{"key":"2022020121021980800_DKU568C17","doi-asserted-by":"crossref","first-page":"508","DOI":"10.1128\/JB.181.2.508-520.1999","article-title":"Localization of FtsI (PBP3) to the septal ring requires its membrane anchor, the Z ring, FtsA, FtsQ, and FtsL","volume":"181","author":"Weiss","year":"1999","journal-title":"J Bacteriol"},{"key":"2022020121021980800_DKU568C18","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1046\/j.1365-2958.1997.5041869.x","article-title":"Localization of the Escherichia coli cell division protein Ftsl (PBP3) to the division site and cell pole","volume":"25","author":"Weiss","year":"1997","journal-title":"Mol Microbiol"},{"key":"2022020121021980800_DKU568C19","volume-title":"Performance Standards for Antimicrobial Susceptibility Testing: Twenty-third Informational Supplement M100-S23","author":"Clinical and Laboratory Standards Institute","year":"2013"},{"key":"2022020121021980800_DKU568C20","doi-asserted-by":"crossref","first-page":"6640","DOI":"10.1073\/pnas.120163297","article-title":"One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products","volume":"97","author":"Datsenko","year":"2000","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022020121021980800_DKU568C21","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1128\/AAC.00756-10","article-title":"Activities of NXL104 combinations with ceftazidime and aztreonam against carbapenemase-producing Enterobacteriaceae","volume":"55","author":"Livermore","year":"2011","journal-title":"Antimicrob Agents Chemother"},{"key":"2022020121021980800_DKU568C22","doi-asserted-by":"crossref","first-page":"1031","DOI":"10.1093\/jac\/dkf240","article-title":"Biochemical analysis of the ceftazidime-hydrolysing extended-spectrum \u03b2-lactamase CTX-M-15 and of its structurally related \u03b2-lactamase CTX-M-3","volume":"50","author":"Poirel","year":"2002","journal-title":"J Antimicrob Chemother"},{"key":"2022020121021980800_DKU568C23","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1042\/bj2550123","article-title":"A survey of the kinetic parameters of class C \u03b2-lactamases. cephalosporins and other \u03b2-lactam compounds","volume":"255","author":"Galleni","year":"1988","journal-title":"Biochem J"},{"key":"2022020121021980800_DKU568C24","doi-asserted-by":"crossref","first-page":"5704","DOI":"10.1128\/AAC.03057-14","article-title":"Avibactam and class C \u03b2-lactamases: Mechanism of inhibition, conservation of binding pocket and implications for resistance","volume":"58","author":"Lahiri","year":"2014","journal-title":"Antimicrob Agents Chemother"},{"key":"2022020121021980800_DKU568C25","doi-asserted-by":"crossref","first-page":"27960","DOI":"10.1074\/jbc.M113.485979","article-title":"Kinetics of avibactam inhibition against class A, C, and D \u03b2-lactamases","volume":"288","author":"Ehmann","year":"2013","journal-title":"J Biol Chem"},{"key":"2022020121021980800_DKU568C26","doi-asserted-by":"crossref","first-page":"11663","DOI":"10.1073\/pnas.1205073109","article-title":"Avibactam is a covalent, reversible, non-\u03b2-lactam \u03b2-lactamase inhibitor","volume":"109","author":"Ehmann","year":"2012","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022020121021980800_DKU568C27","doi-asserted-by":"crossref","first-page":"2312","DOI":"10.1128\/AAC.35.11.2312","article-title":"Comparison of cefepime, cefpirome, and cefaclidine binding affinities for penicillin-binding proteins in Escherichia coli K-12 and Pseudomonas aeruginosa SC8329","volume":"35","author":"Pucci","year":"1991","journal-title":"Antimicrob Agents Chemother"},{"key":"2022020121021980800_DKU568C28","doi-asserted-by":"crossref","first-page":"1510","DOI":"10.1128\/AAC.01529-07","article-title":"Affinity of doripenem and comparators to penicillin-binding proteins in Escherichia coli and Pseudomonas aeruginosa","volume":"52","author":"Davies","year":"2008","journal-title":"Antimicrob Agents Chemother"},{"key":"2022020121021980800_DKU568C29","doi-asserted-by":"crossref","first-page":"521","DOI":"10.7164\/antibiotics.45.521","article-title":"RU 29 246, the active compound of the cephalosporin-prodrug-ester HR 916. II. Stability to \u03b2-lactamases and affinity for penicillin-binding proteins","volume":"45","author":"Markus","year":"1992","journal-title":"J Antibiot (Tokyo)"},{"key":"2022020121021980800_DKU568C30","doi-asserted-by":"crossref","first-page":"e98042","DOI":"10.1371\/journal.pone.0098042","article-title":"Crystal structure of penicillin-binding protein 3 (PBP3) from Escherichia coli","volume":"9","author":"Sauvage","year":"2014","journal-title":"PLoS One"},{"key":"2022020121021980800_DKU568C31","doi-asserted-by":"crossref","first-page":"20536","DOI":"10.1021\/ja208835z","article-title":"Distinctive attributes of \u03b2-lactam target proteins in Acinetobacter baumannii relevant to development of new antibiotics","volume":"133","author":"Han","year":"2011","journal-title":"J Am Chem Soc"},{"key":"2022020121021980800_DKU568C32","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1016\/j.diagmicrobio.2014.07.007","article-title":"Dual \u03b2-lactam therapy for serious Gram-negative infections: is it time to revisit?","volume":"80","author":"Rahme","year":"2014","journal-title":"Diag Microbiol Infect Dis"},{"key":"2022020121021980800_DKU568C33","doi-asserted-by":"crossref","first-page":"22002","DOI":"10.1073\/pnas.1013092107","article-title":"Structural basis for effectiveness of siderophore-conjugated monocarbams against clinically relevant strains of Pseudomonas aeruginosa","volume":"107","author":"Han","year":"2010","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022020121021980800_DKU568C34","volume-title":"Breakpoint Tables for Interpretation of MICs and Zone Diameters","author":"EUCAST","year":"2015"}],"container-title":["Journal of Antimicrobial Chemotherapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jac\/article-pdf\/70\/5\/1420\/42355049\/jac_70_5_1420.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jac\/article-pdf\/70\/5\/1420\/42355049\/jac_70_5_1420.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,1]],"date-time":"2022-02-01T16:10:37Z","timestamp":1643731837000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jac\/article\/70\/5\/1420\/691381"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,1,28]]},"references-count":34,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2015,1,28]]},"published-print":{"date-parts":[[2015,5,1]]}},"URL":"https:\/\/doi.org\/10.1093\/jac\/dku568","relation":{},"ISSN":["1460-2091","0305-7453"],"issn-type":[{"value":"1460-2091","type":"electronic"},{"value":"0305-7453","type":"print"}],"subject":[],"published-other":{"date-parts":[[2015,5]]},"published":{"date-parts":[[2015,1,28]]}}}