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| | ==Crystal structure of E.coli Multidrug/H+ antiporter MdfA in outward open conformation with bound Fab fragment== | | ==Crystal structure of E.coli Multidrug/H+ antiporter MdfA in outward open conformation with bound Fab fragment== |
| - | <StructureSection load='6gv1' size='340' side='right' caption='[[6gv1]], [[Resolution|resolution]] 3.40Å' scene=''> | + | <StructureSection load='6gv1' size='340' side='right'caption='[[6gv1]], [[Resolution|resolution]] 3.40Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[6gv1]] is a 3 chain structure with sequence from [http://en.wikipedia.org/wiki/Ecoli Ecoli] and [http://en.wikipedia.org/wiki/Lk3_transgenic_mice Lk3 transgenic mice]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6GV1 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6GV1 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[6gv1]] is a 3 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli_K-12 Escherichia coli K-12] and [https://en.wikipedia.org/wiki/Mus_musculus Mus musculus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6GV1 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6GV1 FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 3.4Å</td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6gv1 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6gv1 OCA], [http://pdbe.org/6gv1 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6gv1 RCSB], [http://www.ebi.ac.uk/pdbsum/6gv1 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6gv1 ProSAT]</span></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> |
| | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=6gv1 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6gv1 OCA], [https://pdbe.org/6gv1 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6gv1 RCSB], [https://www.ebi.ac.uk/pdbsum/6gv1 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6gv1 ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/MDFA_ECOLI MDFA_ECOLI]] Efflux pump driven by the proton motive force. Confers resistance to a broad spectrum of chemically unrelated drugs. Confers resistance to a diverse group of cationic or zwitterionic lipophilic compounds such as ethidium bromide, tetraphenylphosphonium, rhodamine, daunomycin, benzalkonium, rifampicin, tetracycline, puromycin, and to chemically unrelated, clinically important antibiotics such as chloramphenicol, erythromycin, and certain aminoglycosides and fluoroquinolones. Overexpression results in isopropyl-beta-D-thiogalactopyranoside (IPTG) exclusion and spectinomycin sensitivity. Transport of neutral substrates is electrogenic, whereas transport of cationic substrates is electroneutral. In addition to its role in multidrug resistance, confers extreme alkaline pH resistance, allowing the growth under conditions that are close to those used normally by alkaliphiles. This activity requires Na(+) or K(+).<ref>PMID:12578981</ref> <ref>PMID:15371593</ref> <ref>PMID:9079913</ref> <ref>PMID:9644262</ref> <ref>PMID:9811673</ref> | + | [https://www.uniprot.org/uniprot/MDFA_ECOLI MDFA_ECOLI] Efflux pump driven by the proton motive force. Confers resistance to a broad spectrum of chemically unrelated drugs. Confers resistance to a diverse group of cationic or zwitterionic lipophilic compounds such as ethidium bromide, tetraphenylphosphonium, rhodamine, daunomycin, benzalkonium, rifampicin, tetracycline, puromycin, and to chemically unrelated, clinically important antibiotics such as chloramphenicol, erythromycin, and certain aminoglycosides and fluoroquinolones. Overexpression results in isopropyl-beta-D-thiogalactopyranoside (IPTG) exclusion and spectinomycin sensitivity. Transport of neutral substrates is electrogenic, whereas transport of cationic substrates is electroneutral. In addition to its role in multidrug resistance, confers extreme alkaline pH resistance, allowing the growth under conditions that are close to those used normally by alkaliphiles. This activity requires Na(+) or K(+).<ref>PMID:12578981</ref> <ref>PMID:15371593</ref> <ref>PMID:9079913</ref> <ref>PMID:9644262</ref> <ref>PMID:9811673</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Ecoli]] | + | [[Category: Escherichia coli K-12]] |
| - | [[Category: Lk3 transgenic mice]]
| + | [[Category: Large Structures]] |
| - | [[Category: Nagarathinam, K]]
| + | [[Category: Mus musculus]] |
| - | [[Category: Parthier, C]] | + | [[Category: Nagarathinam K]] |
| - | [[Category: Stubbs, M T]] | + | [[Category: Parthier C]] |
| - | [[Category: Tanabe, M]] | + | [[Category: Stubbs MT]] |
| - | [[Category: Drug proton antiporter]] | + | [[Category: Tanabe M]] |
| - | [[Category: Major facilitator superfamily]] | + | |
| - | [[Category: Mfs transporter]] | + | |
| - | [[Category: Multidrug resistance]]
| + | |
| - | [[Category: Proton transport]]
| + | |
| - | [[Category: Transport protein]]
| + | |
| Structural highlights
Function
MDFA_ECOLI Efflux pump driven by the proton motive force. Confers resistance to a broad spectrum of chemically unrelated drugs. Confers resistance to a diverse group of cationic or zwitterionic lipophilic compounds such as ethidium bromide, tetraphenylphosphonium, rhodamine, daunomycin, benzalkonium, rifampicin, tetracycline, puromycin, and to chemically unrelated, clinically important antibiotics such as chloramphenicol, erythromycin, and certain aminoglycosides and fluoroquinolones. Overexpression results in isopropyl-beta-D-thiogalactopyranoside (IPTG) exclusion and spectinomycin sensitivity. Transport of neutral substrates is electrogenic, whereas transport of cationic substrates is electroneutral. In addition to its role in multidrug resistance, confers extreme alkaline pH resistance, allowing the growth under conditions that are close to those used normally by alkaliphiles. This activity requires Na(+) or K(+).[1] [2] [3] [4] [5]
Publication Abstract from PubMed
Multidrug resistance (MDR) poses a major challenge to medicine. A principle cause of MDR is through active efflux by MDR transporters situated in the bacterial membrane. Here we present the crystal structure of the major facilitator superfamily (MFS) drug/H(+) antiporter MdfA from Escherichia coli in an outward open conformation. Comparison with the inward facing (drug binding) state shows that, in addition to the expected change in relative orientations of the N- and C-terminal lobes of the antiporter, the conformation of TM5 is kinked and twisted. In vitro reconstitution experiments demonstrate the importance of selected residues for transport and molecular dynamics simulations are used to gain insights into antiporter switching. With the availability of structures of alternative conformational states, we anticipate that MdfA will serve as a model system for understanding drug efflux in MFS MDR antiporters.
Outward open conformation of a Major Facilitator Superfamily multidrug/H(+) antiporter provides insights into switching mechanism.,Nagarathinam K, Nakada-Nakura Y, Parthier C, Terada T, Juge N, Jaenecke F, Liu K, Hotta Y, Miyaji T, Omote H, Iwata S, Nomura N, Stubbs MT, Tanabe M Nat Commun. 2018 Oct 1;9(1):4005. doi: 10.1038/s41467-018-06306-x. PMID:30275448[6]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Lewinson O, Adler J, Poelarends GJ, Mazurkiewicz P, Driessen AJ, Bibi E. The Escherichia coli multidrug transporter MdfA catalyzes both electrogenic and electroneutral transport reactions. Proc Natl Acad Sci U S A. 2003 Feb 18;100(4):1667-72. Epub 2003 Feb 10. PMID:12578981 doi:http://dx.doi.org/10.1073/pnas.0435544100
- ↑ Lewinson O, Padan E, Bibi E. Alkalitolerance: a biological function for a multidrug transporter in pH homeostasis. Proc Natl Acad Sci U S A. 2004 Sep 28;101(39):14073-8. Epub 2004 Sep 15. PMID:15371593 doi:http://dx.doi.org/10.1073/pnas.0405375101
- ↑ Edgar R, Bibi E. MdfA, an Escherichia coli multidrug resistance protein with an extraordinarily broad spectrum of drug recognition. J Bacteriol. 1997 Apr;179(7):2274-80. PMID:9079913
- ↑ Mine T, Morita Y, Kataoka A, Mizushima T, Tsuchiya T. Evidence for chloramphenicol/H+ antiport in Cmr (MdfA) system of Escherichia coli and properties of the antiporter. J Biochem. 1998 Jul;124(1):187-93. PMID:9644262
- ↑ Bohn C, Bouloc P. The Escherichia coli cmlA gene encodes the multidrug efflux pump Cmr/MdfA and is responsible for isopropyl-beta-D-thiogalactopyranoside exclusion and spectinomycin sensitivity. J Bacteriol. 1998 Nov;180(22):6072-5. PMID:9811673
- ↑ Nagarathinam K, Nakada-Nakura Y, Parthier C, Terada T, Juge N, Jaenecke F, Liu K, Hotta Y, Miyaji T, Omote H, Iwata S, Nomura N, Stubbs MT, Tanabe M. Outward open conformation of a Major Facilitator Superfamily multidrug/H(+) antiporter provides insights into switching mechanism. Nat Commun. 2018 Oct 1;9(1):4005. doi: 10.1038/s41467-018-06306-x. PMID:30275448 doi:http://dx.doi.org/10.1038/s41467-018-06306-x
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