{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T22:15:10Z","timestamp":1791152110982,"version":"4.1.0"},"reference-count":25,"publisher":"Wiley","issue":"14","license":[{"start":{"date-parts":[[2010,7,6]],"date-time":"2010-07-06T00:00:00Z","timestamp":1278374400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Overseas Research Studentship"},{"name":"Cambridge Commonwealth Trust Scholarship"},{"name":"American Alumni Award and Charter Studentship from St. Edmund's College, Cambridge"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Proteins"],"published-print":{"date-parts":[[2010,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Recent crystal structures of the multidrug ATP\u2010binding cassette (ABC) exporters Sav1866 from <jats:italic>Staphylococcus aureus<\/jats:italic>, MsbA from <jats:italic>Escherichia coli<\/jats:italic>, <jats:italic>Vibrio cholera<\/jats:italic>, and <jats:italic>Salmonella typhimurium<\/jats:italic>, and mouse ABCB1a suggest a common alternating access mechanism for export. However, the molecular framework underlying this mechanism is critically dependent on assumed conformational relationships between nonidentical crystal structures and therefore requires biochemical verification. The structures of homodimeric MsbA reveal a pair of glutamate residues (E208 and E208\u2032) in the intracellular domains of its two half\u2010transporters, close to the nucleotide\u2010binding domains (NBDs), which are in close proximity of each other in the outward\u2010facing state but not in the inward\u2010facing state. Using intermolecular cysteine crosslinking between E208C and E208C\u2032 in <jats:italic>E. coli<\/jats:italic> MsbA, we demonstrate that the NBDs dissociate in nucleotide\u2010free conditions and come close on ATP binding and ADP\u00b7vanadate trapping. Interestingly, ADP alone separates the half\u2010transporters like a nucleotide\u2010free state, presumably for the following catalytic cycle. Our data fill persistent gaps in current studies on the conformational dynamics of a variety of ABC exporters. Based on a single biochemical method, the findings describe a conformational cycle for a single ABC exporter at major checkpoints of the ATPase reaction under experimental conditions, where the exporter is transport active. Proteins 2010. \u00a9 2010 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/prot.22813","type":"journal-article","created":{"date-parts":[[2010,7,7]],"date-time":"2010-07-07T08:47:01Z","timestamp":1278492421000},"page":"2867-2872","source":"Crossref","is-referenced-by-count":35,"title":["Dissection of the conformational cycle of the multidrug\/lipidA ABC exporter MsbA"],"prefix":"10.1002","volume":"78","author":[{"given":"Rupak","family":"Doshi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Woebking","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hendrik W.","family":"van Veen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2010,9,8]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1194\/jlr.R800060-JLR200"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi800778d"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.18.6363-6369.2005"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M306226200"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi8006737"},{"key":"e_1_2_6_7_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M205857200"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M708274200"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20081364"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi801745u"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0709388104"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2008.11.006"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2009.08.051"},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2009.08.050"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0050271"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2009.12.4193"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2009.07.009"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature05155"},{"key":"e_1_2_6_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1097-2765(02)00576-2"},{"key":"e_1_2_6_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2007.01.073"},{"key":"e_1_2_6_21_2","volume-title":"The PyMOL molecular graphics system, version 1.2rl","author":"Delano WL","year":"2009"},{"key":"e_1_2_6_22_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1168750"},{"key":"e_1_2_6_23_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.056432"},{"key":"e_1_2_6_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bcp.2007.10.022"},{"key":"e_1_2_6_25_2","doi-asserted-by":"crossref","first-page":"3662","DOI":"10.1128\/aem.62.10.3662-3667.1996","article-title":"Controlled gene expression systems for Lactococcus lactis with the food\u2010 grade inducer nisin","volume":"62","author":"de Ruyter PG","year":"1996","journal-title":"Appl Environ Microbiol"},{"key":"e_1_2_6_26_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M707028200"}],"container-title":["Proteins: Structure, Function, and Bioinformatics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fprot.22813","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fprot.22813","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/prot.22813","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,9]],"date-time":"2023-10-09T22:13:34Z","timestamp":1696889614000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/prot.22813"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2010,9,8]]},"references-count":25,"journal-issue":{"issue":"14","published-print":{"date-parts":[[2010,11]]}},"alternative-id":["10.1002\/prot.22813"],"URL":"https:\/\/doi.org\/10.1002\/prot.22813","archive":["Portico"],"relation":{},"ISSN":["0887-3585","1097-0134"],"issn-type":[{"value":"0887-3585","type":"print"},{"value":"1097-0134","type":"electronic"}],"subject":[],"published":{"date-parts":[[2010,9,8]]}}}