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Although Isoxazolyl Penicillins have been available since the 1970s, there are currently no structures in complex with class-A\n                    <jats:italic>\u03b2<\/jats:italic>\n                    -lactamases available. Here we have analysed the structure of the clinically relevant\n                    <jats:italic>\u03b2<\/jats:italic>\n                    -lactamase CTX-M-14 from\n                    <jats:italic>Klebsiella pneumoniae<\/jats:italic>\n                    near physiological temperatures, via serial synchrotron crystallography. We demonstrate the acyl-enzyme intermediates of the catalytically impaired CTX-M-14 mutant E166A in complex with three Isoxazolyl-Penicillins: Oxacillin, Cloxacillin and Dicloxacillin. Structural comparisons of CTX-M-Penicillin complexes show that, while conserved active-site interactions are maintained, each Isoxazolyl-Penicillin adopts a distinct conformation. While the three derivatives differ only by one and two chlorine atoms, respectively, their conformational heterogeneity appears to be increased by chlorination of the phenyl ring.\n                  <\/jats:p>","DOI":"10.1038\/s42004-025-01801-x","type":"journal-article","created":{"date-parts":[[2025,12,1]],"date-time":"2025-12-01T16:35:32Z","timestamp":1764606932000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Binding mode of Isoxazolyl Penicillins to a Class-A \u03b2-lactamase at ambient conditions"],"prefix":"10.1038","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-0585-7038","authenticated-orcid":false,"given":"Gargi","family":"Gore","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2738-7682","authenticated-orcid":false,"given":"Andreas","family":"Prester","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"von Stetten","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim","family":"Bartels","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1685-8605","authenticated-orcid":false,"given":"Eike C.","family":"Schulz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,12,1]]},"reference":[{"key":"1801_CR1","doi-asserted-by":"publisher","first-page":"2221","DOI":"10.1016\/S0140-6736(22)02185-7","volume":"400","author":"KS Ikuta","year":"2022","unstructured":"Ikuta, K. 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