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Here, we quantified the uptake of kanamycin across the major porin channels OmpF and OmpC present in the outer membrane of Escherichia coli. Our analysis revealed that, despite its relatively large size, about 10\u201320 kanamycin molecules per second permeate through OmpF and OmpC under a 10 \u03bcM concentration gradient, whereas OmpN does not allow the passage. Molecular simulations elucidate the uptake mechanism of kanamycin through these porins. Whole-cell studies with a defined set of E.\u00a0coli porin mutants provide evidence that translocation of kanamycin via porins is relevant for antibiotic potency. The values are discussed with respect to other antibiotics.<\/jats:p>","DOI":"10.1021\/acsinfecdis.0c00102","type":"journal-article","created":{"date-parts":[[2020,5,5]],"date-time":"2020-05-05T17:41:56Z","timestamp":1588700516000},"page":"1855-1865","source":"Crossref","is-referenced-by-count":72,"title":["Kanamycin Uptake into\n                    <i>Escherichia coli<\/i>\n                    Is Facilitated by OmpF and OmpC Porin Channels Located in the Outer\nMembrane"],"prefix":"10.1021","volume":"6","author":[{"given":"Jayesh\nArun","family":"Bafna","sequence":"first","affiliation":[{"name":"Jacobs University Bremen , , ,","place":["Bremen, Germany, D-28719"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eula\u0300lia","family":"Sans-Serramitjana","sequence":"additional","affiliation":[{"name":"Jacobs University Bremen , , ,","place":["Bremen, Germany, D-28719"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0122-9769","authenticated-orcid":true,"given":"Silvia","family":"Acosta-Gutie\u0301rrez","sequence":"additional","affiliation":[{"name":"University College London , , ,","place":["London, United Kingdom, WC1E 6BT"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1141-1449","authenticated-orcid":true,"given":"Igor V.","family":"Bodrenko","sequence":"additional","affiliation":[{"name":"IOM\/CNR, Sezione di Cagliari, Cittadella Universitaria di Monserrato , ,","place":["Monserrato, Italy, 09042"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5236-8443","authenticated-orcid":true,"given":"Daniel","family":"Ho\u0308ro\u0308mpo\u0308li","sequence":"additional","affiliation":[{"name":"University of Tu\u0308bingen , , ,","place":["Tu\u0308bingen, Germany, D-72076"]},{"name":"German Center for Infection Research (DZIF) Partner Site , ,","place":["Tu\u0308bingen, Germany, D-72076"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1585-8715","authenticated-orcid":true,"given":"Anne","family":"Berscheid","sequence":"additional","affiliation":[{"name":"University of Tu\u0308bingen , , ,","place":["Tu\u0308bingen, Germany, D-72076"]},{"name":"German Center for Infection Research (DZIF) Partner Site , ,","place":["Tu\u0308bingen, Germany, D-72076"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9364-1832","authenticated-orcid":true,"given":"Heike","family":"Bro\u0308tz-Oesterhelt","sequence":"additional","affiliation":[{"name":"University of Tu\u0308bingen , , ,","place":["Tu\u0308bingen, Germany, D-72076"]},{"name":"German Center for Infection Research (DZIF) Partner Site , ,","place":["Tu\u0308bingen, Germany, D-72076"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathias","family":"Winterhalter","sequence":"additional","affiliation":[{"name":"Jacobs University Bremen , , ,","place":["Bremen, Germany, D-28719"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4272-902X","authenticated-orcid":true,"given":"Matteo","family":"Ceccarelli","sequence":"additional","affiliation":[{"name":"IOM\/CNR, Sezione di Cagliari, Cittadella Universitaria di Monserrato , ,","place":["Monserrato, Italy, 09042"]},{"name":"University of Cagliari, and CNR\/IOM, Cittadella Universitaria di Monserrato , , ,","place":["Monserrato, Italy, 09042"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,5,5]]},"reference":[{"key":"2026081722190598200_cit1","doi-asserted-by":"publisher","first-page":"593","DOI":"10.1128\/MMBR.67.4.593-656.2003","article-title":"Molecular\nBasis of Bacterial Outer Membrane Permeability Revisited","volume":"67","author":"Nikaido","year":"2003","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"2026081722190598200_cit2","doi-asserted-by":"publisher","first-page":"299","DOI":"10.1038\/nature22308","article-title":"Predictive\nCompound Accumulation\nRules Yield a Broad-Spectrum Antibiotic","volume":"545","author":"Richter","year":"2017","journal-title":"Nature"},{"key":"2026081722190598200_cit3","doi-asserted-by":"publisher","first-page":"164","DOI":"10.1038\/s41579-019-0294-2","article-title":"Porins and Small-Molecule Translocation\nacross the\nOuter Membrane of Gram-Negative Bacteria","volume":"18","author":"Vergalli","year":"2020","journal-title":"Nat.\nRev. Microbiol."},{"key":"2026081722190598200_cit4","doi-asserted-by":"publisher","first-page":"877","DOI":"10.1016\/0006-291X(76)90913-X","article-title":"Identification\nof the Outer Membrane Protein of E. Coli That Produces Transmembrane\nChannels in Reconstituted Vesicle Membranes","volume":"71","author":"Nakae","year":"1976","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"2026081722190598200_cit5","doi-asserted-by":"publisher","first-page":"1688","DOI":"10.1021\/acsinfecdis.9b00256","article-title":"Size Matters and How\nYou Measure It: A Gram-Negative\nAntibacterial Example Exceeding Typical Molecular Weight Limits","volume":"5","author":"Ruggiu","year":"2019","journal-title":"ACS Infect. Dis."},{"key":"2026081722190598200_cit6","doi-asserted-by":"publisher","first-page":"1487","DOI":"10.1021\/acsinfecdis.8b00108","article-title":"Getting Drugs into Gram-Negative\nBacteria: Rational Rules for Permeation through General Porins","volume":"4","author":"Acosta-Gutie\u0301rrez","year":"2018","journal-title":"ACS Infect. Dis."},{"key":"2026081722190598200_cit7","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0025825","article-title":"Altered Antibiotic\nTransport in OmpC Mutants Isolated from a Series of Clinical Strains\nof Multi-Drug Resistant E. 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Phys."},{"key":"2026081722190598200_cit21","doi-asserted-by":"publisher","first-page":"5465","DOI":"10.1021\/acsnano.6b08613","article-title":"Bacterial Outer Membrane Porins as\nElectrostatic Nanosieves: Exploring Transport Rules of Small Polar\nMolecules","volume":"11","author":"Bajaj","year":"2017","journal-title":"ACS Nano"},{"key":"2026081722190598200_cit22","doi-asserted-by":"publisher","first-page":"76","DOI":"10.1016\/j.str.2012.10.014","article-title":"The Binding of Antibiotics\nin OmpF Porin","volume":"21","author":"Ziervogel","year":"2013","journal-title":"Structure"},{"key":"2026081722190598200_cit23","doi-asserted-by":"publisher","DOI":"10.1038\/msb4100050","article-title":"Construction\nof Escherichia Coli K-12 In-frame, Single-gene Knockout Mutants: The\nKeio Collection","author":"Baba","year":"2006","journal-title":"Mol. Syst. 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Agents Chemother."},{"key":"2026081722190598200_cit27","doi-asserted-by":"publisher","DOI":"10.1128\/ecosalplus.ESP-0002-2018","article-title":"Aminoglycoside Revival:\nReview of a Historically Important Class of Antimicrobials Undergoing\nRejuvenation","author":"Serio","year":"2018","journal-title":"EcoSal Plus"},{"key":"2026081722190598200_cit28","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1111\/nyas.13598","article-title":"The Challenge\nof Converting Gram-Positive-Only Compounds\ninto Broad-Spectrum Antibiotics","volume":"1435","author":"Richter","year":"2019","journal-title":"Ann. N. Y.\nAcad. Sci."},{"key":"2026081722190598200_cit29","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1039\/C5MD00344J","article-title":"Mechanisms of Resistance to Aminoglycoside Antibiotics:\nOverview and Perspectives","volume":"7","author":"Garneau-Tsodikova","year":"2016","journal-title":"MedChemComm"},{"key":"2026081722190598200_cit30","doi-asserted-by":"publisher","first-page":"1295","DOI":"10.1021\/acs.jpclett.7b00062","article-title":"A General Method to\nDetermine the Flux of Charged Molecules Through Nanopores Applied\nto SS-Lactamase Inhibitors and OmpF","volume":"8","author":"Ghai","year":"2017","journal-title":"J. Phys.\nChem. Lett."},{"key":"2026081722190598200_cit31","doi-asserted-by":"publisher","first-page":"7030","DOI":"10.1074\/jbc.RA117.000705","article-title":"Ampicillin Permeation across OmpF,\nthe Major Outer-Membrane Channel in Escherichia Coli","volume":"293","author":"Ghai","year":"2018","journal-title":"J. Biol. 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Biomol.\nScreening"},{"key":"2026081722190598200_cit33","doi-asserted-by":"publisher","first-page":"1184","DOI":"10.1021\/acssensors.7b00311","article-title":"Sensing Single Molecule Penetration into Nanopores:\nPushing the Time Resolution to the Diffusion Limit","volume":"2","author":"Bodrenko","year":"2017","journal-title":"ACS Sensors"},{"key":"2026081722190598200_cit34","doi-asserted-by":"publisher","first-page":"58","DOI":"10.1093\/jac\/dky396","article-title":"Antibiotics\nand Efflux: Combined Spectrofluorimetry\nand Mass Spectrometry to Evaluate the Involvement of Concentration\nand Efflux Activity in Antibiotic Intracellular Accumulation","volume":"74","author":"Dumont","year":"2018","journal-title":"J. Antimicrob. 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Chem."},{"key":"2026081722190598200_cit46","doi-asserted-by":"publisher","first-page":"299","DOI":"10.1016\/S0163-7258(03)00037-8","article-title":"Formation\nand Plasticity of GABAergic Synapses: Physiological\nMechanisms and Pathophysiological Implications","volume":"98","author":"Fritschy","year":"2003","journal-title":"Pharmacol. Ther."},{"key":"2026081722190598200_cit47","doi-asserted-by":"publisher","first-page":"247260","DOI":"10.1016\/j.jmgm.2005.12.005","article-title":"Automatic\natom type\nand bond type perception in molecular mechanical calculations","volume":"25","author":"Wang","year":"2006","journal-title":"J. Mol. 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