{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T22:29:27Z","timestamp":1787005767051,"version":"build-2736575974"},"reference-count":67,"publisher":"National Academy of Sciences","issue":"44","license":[{"start":{"date-parts":[[2024,10,23]],"date-time":"2024-10-23T00:00:00Z","timestamp":1729641600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["82272379"],"award-info":[{"award-number":["82272379"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100012166","name":"National Key R&D Program of China","doi-asserted-by":"crossref","award":["2020YFA0907"],"award-info":[{"award-number":["2020YFA0907"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project","award":["TSBICIP-KJGG-014"],"award-info":[{"award-number":["TSBICIP-KJGG-014"]}]},{"DOI":"10.13039\/501100021171","name":"Guangdong Basic and Applied Basic Research Foundation","doi-asserted-by":"crossref","award":["2019A1515110675"],"award-info":[{"award-number":["2019A1515110675"]}],"id":[{"id":"10.13039\/501100021171","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100002858","name":"China Postdoctoral Science Foundation","doi-asserted-by":"publisher","award":["2019M660048"],"award-info":[{"award-number":["2019M660048"]}],"id":[{"id":"10.13039\/501100002858","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2024,10,29]]},"abstract":"<jats:p>\n                    EfpA, the first major facilitator superfamily (MFS) protein identified in\n                    <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                    (Mtb), is an essential efflux pump implicated in resistance to multiple drugs. EfpA-inhibitors have been developed to kill drug-tolerant Mtb. However, the biological function of EfpA has not yet been elucidated. Here, we present the cryo-EM structures of EfpA complexed with lipids or the inhibitor BRD-8000.3 at resolutions of 2.9 \u00c5 and 3.4 \u00c5, respectively. Unexpectedly, EfpA forms an antiparallel dimer. Functional studies reveal that EfpA is a lipid transporter and BRD-8000.3 inhibits its lipid transport activity. Intriguingly, the mutation V319F, known to confer resistance to BRD-8000.3, alters the expression level and oligomeric state of EfpA. Based on our results and the observation of other antiparallel dimers in the MFS family, we propose an antiparallel-function model of EfpA. Collectively, our work provides structural and functional insights into EfpA\u2019s role in lipid transport and drug resistance, which would accelerate the development of antibiotics against this promising drug target.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2412653121","type":"journal-article","created":{"date-parts":[[2024,10,23]],"date-time":"2024-10-23T11:45:51Z","timestamp":1729683951000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":4,"title":["Structure and function of\n                    <i>Mycobacterium tuberculosis<\/i>\n                    EfpA as a lipid transporter and its inhibition by BRD-8000.3"],"prefix":"10.1073","volume":"121","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7967-1581","authenticated-orcid":false,"given":"Delin","family":"Li","sequence":"first","affiliation":[{"name":"Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"},{"name":"Shanxi Academy of Advanced Research and Innovation, Taiyuan 030032, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaokang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanhang","family":"Yao","sequence":"additional","affiliation":[{"name":"College of Future Technology, Peking University, Beijing 100871, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xue","family":"Sun","sequence":"additional","affiliation":[{"name":"First School of Clinical Medicine, Peking University Health Science Center, Peking University, Beijing 100871, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junqing","family":"Sun","sequence":"additional","affiliation":[{"name":"Shanxi Academy of Advanced Research and Innovation, Taiyuan 030032, China"},{"name":"Shanxi Agricultural University, Jinzhong, Shanxi 030801, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-0458-5132","authenticated-orcid":false,"given":"Xiaomin","family":"Ma","sequence":"additional","affiliation":[{"name":"Cryo-EM Center, Southern University of Science and Technology, Shenzhen 518055, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Yuan","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guijie","family":"Bai","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuefei","family":"Pang","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rongmao","family":"Hua","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tianling","family":"Guo","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuqian","family":"Mi","sequence":"additional","affiliation":[{"name":"Shanxi Academy of Advanced Research and Innovation, Taiyuan 030032, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingzhi","family":"Wu","sequence":"additional","affiliation":[{"name":"College of Future Technology, Peking University, Beijing 100871, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Zhang","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2734-0673","authenticated-orcid":false,"given":"Yan","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Pathogen Microbiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingxia","family":"Liu","sequence":"additional","affiliation":[{"name":"Shenzhen Key Laboratory of Pathogen and Immunity, Shenzhen Third People\u2019s Hospital, Second Hospital Affiliated to Southern University of Science and Technology, Shenzhen 518112, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7737-181X","authenticated-orcid":false,"given":"Peiyi","family":"Wang","sequence":"additional","affiliation":[{"name":"Cryo-EM Center, Southern University of Science and Technology, Shenzhen 518055, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catherine C. L.","family":"Wong","sequence":"additional","affiliation":[{"name":"First School of Clinical Medicine, Peking University Health Science Center, Peking University, Beijing 100871, China"},{"name":"State Key Laboratory of Complex, Severe and Rare Diseases, Clinical Research Institute, Peking Union Medical College Hospital, Chinese Academy of Medical Science &amp; Peking Union Medical College, Beijing 100730, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6466-6248","authenticated-orcid":false,"given":"Xiao-wei","family":"Chen","sequence":"additional","affiliation":[{"name":"College of Future Technology, Peking University, Beijing 100871, China"},{"name":"State Key Laboratory of Membrane Biology, Peking University, Center for Life Sciences, Peking University, Beijing 100871, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0032-5430","authenticated-orcid":false,"given":"Haixia","family":"Xiao","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3869-615X","authenticated-orcid":false,"given":"George Fu","family":"Gao","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"},{"name":"Chinese Academy of Sciences Key Laboratory of Pathogen Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0670-010X","authenticated-orcid":false,"given":"Feng","family":"Gao","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2024,10,23]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/S2666-5247(22)00359-7"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cimid.2020.101574"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1080\/07853890701195262"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/cdli.4.1.23-32.1997"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00232-017-9982-x"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.0030087"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0034538"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.96.22.12833"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00262-21"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.add5907"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02133-16"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.06.033"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0966-842X(97)01064-0"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-micro-091014-104121"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.GPP3-0027-2018"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2013.11.011"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1515\/hsz-2019-0326"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00550-09"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-42073-0"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.01.003"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrev.0c00983"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aba9102"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1106753"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-1315-z"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.9b00373"},{"key":"e_1_3_4_26_2","article-title":"Structures of the essential efflux pump EfpA from Mycobacterium tuberculosis reveal the mechanisms of substrate transport and small-molecule inhibition","author":"Wang S.","year":"2024","unstructured":"S. Wang , Structures of the essential efflux pump EfpA from Mycobacterium tuberculosis reveal the mechanisms of substrate transport and small-molecule inhibition. Res Sq [Preprint] 2024).","journal-title":"Res Sq [Preprint]"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-1923-7"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1038\/341456a0"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb1057"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-05530-2"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1088196"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1311485110"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2012.157"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/21.9.2107"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M116.765719"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-1963-z"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature11524"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature13306"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2023113418"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41594-020-0425-5"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03782-y"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2006.05174.x"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-28361-1"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0173126"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/cdf571"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature20820"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00030a011"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.306860"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03579-z"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1042\/BST20160217"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/20.12.3056"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature13074"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4169"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444910007493"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.20084"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444994003112"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444902016657"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909042073"},{"key":"e_1_3_4_59_2","unstructured":"L. L. C. Schrodinger The PyMOL Molecular Graphics System. Version 1.8 (2015)."},{"key":"e_1_3_4_60_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1002708"},{"key":"e_1_3_4_61_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4519"},{"key":"e_1_3_4_62_2","doi-asserted-by":"publisher","DOI":"10.1002\/0471250953.bi0313s48"},{"key":"e_1_3_4_63_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2022.101333"},{"key":"e_1_3_4_64_2","doi-asserted-by":"crossref","unstructured":"F. Gao X. Zhang D. Li X. Ma Structure of the multidrug efflux pump EfpA from M. tuberculosis complexed with lipids. Electron Microscopy Data Bank. https:\/\/www.ebi.ac.uk\/emdb\/EMD-38093. Deposited 22 November 2023.","DOI":"10.2210\/pdb8x6x\/pdb"},{"key":"e_1_3_4_65_2","unstructured":"D. Li J. Sun The structure of EfpA_BRD-8000.3 complex. Electron Microscopy Data Bank. https:\/\/www.ebi.ac.uk\/emdb\/EMD-39432. Deposited 12 March 2024."},{"key":"e_1_3_4_66_2","doi-asserted-by":"crossref","unstructured":"F. Gao X. Zhang D. Li X. Ma Structure of the multidrug efflux pump EfpA from M. tuberculosis complexed with lipids. RCSB Protein Data Bank. https:\/\/www.rcsb.org\/structure\/unreleased\/8X6X. Deposited 22 November 2023.","DOI":"10.2210\/pdb8x6x\/pdb"},{"key":"e_1_3_4_67_2","unstructured":"D. Li J. Sun The structure of EfpA_BRD-8000.3 complex. RCSB Protein Data Bank. https:\/\/www.rcsb.org\/structure\/unreleased\/8YNZ. Deposited 12 March 2024."}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2412653121","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,11,29]],"date-time":"2024-11-29T23:05:28Z","timestamp":1732921528000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2412653121"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,23]]},"references-count":67,"journal-issue":{"issue":"44","published-print":{"date-parts":[[2024,10,29]]}},"alternative-id":["10.1073\/pnas.2412653121"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2412653121","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,23]]},"assertion":[{"value":"2024-07-29","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-09-18","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-10-23","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2412653121"}}