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NDMs have been exceptionally characterized as lipoproteins anchored to the outer membrane. A bioinformatics study on all sequenced \u03b2-lactamases was performed that revealed a predominance of putative lipidated enzymes in the Class D OXAs. Namely, 60% of the OXA Class D enzymes contain a lipobox sequence in their signal peptide, that is expected to trigger lipidation and membrane anchoring. This contrasts with \u03b2-lactamases from other classes, which are predicted to be mostly soluble proteins. Almost all (&gt;99%) putative lipidated OXAs are present in\n                      <jats:italic>Acinetobacter<\/jats:italic>\n                      spp. Importantly, we further demonstrate that OXA-23 and OXA-24\/40 are lipidated, membrane-bound proteins in\n                      <jats:italic>Acinetobacter baumannii<\/jats:italic>\n                      . In contrast, OXA-48 (commonly produced by Enterobacterales) lacks a lipobox and is a soluble protein. Outer membrane vesicles (OMVs) from\n                      <jats:italic>A. baumannii<\/jats:italic>\n                      cells expressing OXA-23 and OXA-24\/40 contain these enzymes in their active form. Moreover, OXA-loaded OMVs were able to protect\n                      <jats:italic>A. baumannii<\/jats:italic>\n                      ,\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                      , and\n                      <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                      cells susceptible to piperacillin and imipenem. These results permit us to conclude that membrane binding is a bacterial host-specific phenomenon in OXA enzymes. These findings reveal that membrane-bound \u03b2-lactamases are more common than expected and support the hypothesis that OMVs loaded with lipidated \u03b2-lactamases are vehicles for antimicrobial resistance and its dissemination. This advantage could be crucial in polymicrobial infections, in which\n                      <jats:italic>Acinetobacter<\/jats:italic>\n                      spp. are usually involved, and underscore the relevance of identifying the cellular localization of lactamases to better understand their physiology and target them.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        \u03b2-lactamases represent the main mechanism of antimicrobial resistance in Gram-negative pathogens. Their catalytic function (cleaving \u03b2-lactam antibiotics) occurs in the bacterial periplasm, where they are commonly reported as soluble proteins. A bioinformatic analysis reveals a significant number of putative lipidated \u03b2-lactamases, expected to be attached to the outer bacterial membrane. Notably, 60% of Class D OXA \u03b2-lactamases (all from\n                        <jats:italic>Acinetobacter<\/jats:italic>\n                        spp.) are predicted as membrane-anchored proteins. We demonstrate that two clinically relevant carbapenemases, OXA-23 and OXA-24\/40, are membrane-bound proteins in\n                        <jats:italic>A. baumannii<\/jats:italic>\n                        . This cellular localization favors the secretion of these enzymes into outer membrane vesicles that transport them outside the boundaries of the cell. \u03b2-lactamase-loaded vesicles can protect populations of antibiotic-susceptible bacteria, enabling them to thrive in the presence of \u03b2-lactam antibiotics. The ubiquity of this phenomenon suggests that it may have influenced the dissemination of resistance mediated by\n                        <jats:italic>Acinetobacter<\/jats:italic>\n                        spp., particularly in polymicrobial infections, being a potent evolutionary advantage.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.03343-24","type":"journal-article","created":{"date-parts":[[2024,12,13]],"date-time":"2024-12-13T09:01:12Z","timestamp":1734080472000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":30,"title":["OXA \u03b2-lactamases from\n                    <i>Acinetobacter<\/i>\n                    spp. are membrane bound and secreted into outer membrane vesicles"],"prefix":"10.1128","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-5585-9239","authenticated-orcid":false,"given":"Lucia","family":"Capodimonte","sequence":"first","affiliation":[{"name":"Instituto de Biolog\u00eda Molecular y Celular de Rosario (CONICET IBR -UNR)","place":["Rosario, Argentina"]},{"name":"\u00c1rea Biof\u00edsica, Facultad de Ciencias Bioqu\u00edmicas y Farmac\u00e9uticas, Universidad Nacional de Rosario","place":["Rosario, Argentina"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernando Teixeira Pinto","family":"Meireles","sequence":"additional","affiliation":[{"name":"Institute of Bioengineering, School of Life Science, \u00c9cole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL)","place":["Lausanne, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guillermo","family":"Bahr","sequence":"additional","affiliation":[{"name":"Instituto de Biolog\u00eda Molecular y Celular de Rosario (CONICET IBR -UNR)","place":["Rosario, Argentina"]},{"name":"\u00c1rea Biof\u00edsica, Facultad de Ciencias Bioqu\u00edmicas y Farmac\u00e9uticas, Universidad Nacional de Rosario","place":["Rosario, Argentina"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3299-894X","authenticated-orcid":true,"given":"Robert A.","family":"Bonomo","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine","place":["Cleveland, USA"]},{"name":"Research Service, Louis Stokes Cleveland Department of Veterans Affairs Medical Center","place":["Cleveland, USA"]},{"name":"Departments of Pharmacology, Biochemistry, Proteomics and Bioinformatics, Case Western Reserve University School of Medicine","place":["Cleveland, USA"]},{"name":"CWRU-Cleveland VAMC Center for Antimicrobial Resistance and Epidemiology (Case VA CARES)","place":["Cleveland, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matteo","family":"Dal Peraro","sequence":"additional","affiliation":[{"name":"Institute of Bioengineering, School of Life Science, \u00c9cole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL)","place":["Lausanne, Switzerland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9357-3486","authenticated-orcid":false,"given":"Carolina","family":"L\u00f3pez","sequence":"additional","affiliation":[{"name":"Instituto de Biolog\u00eda Molecular y Celular de Rosario (CONICET IBR -UNR)","place":["Rosario, Argentina"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7978-3233","authenticated-orcid":true,"given":"Alejandro J.","family":"Vila","sequence":"additional","affiliation":[{"name":"Instituto de Biolog\u00eda Molecular y Celular de Rosario (CONICET IBR -UNR)","place":["Rosario, Argentina"]},{"name":"\u00c1rea Biof\u00edsica, 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