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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-30372 | |||||||||
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Title | Cryo-EM structure of P.aeruginosa MlaFEBD | |||||||||
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![]() | Mla complex / Lipid transporter / MEMBRANE PROTEIN | |||||||||
Function / homology | ![]() response to nitrite / phospholipid transporter activity / phospholipid transport / ATP-binding cassette (ABC) transporter complex / phospholipid binding / ATP hydrolysis activity / ATP binding / plasma membrane Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.5 Å | |||||||||
![]() | Zhou C / Shi H | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural Insight into Phospholipid Transport by the MlaFEBD Complex from P. aeruginosa. Authors: Changping Zhou / Huigang Shi / Manfeng Zhang / Lijun Zhou / Le Xiao / Shasha Feng / Wonpil Im / Min Zhou / Xinzheng Zhang / Yihua Huang / ![]() ![]() Abstract: The outer membrane (OM) of Gram-negative bacteria, which consists of lipopolysaccharides (LPS) in the outer leaflet and phospholipids (PLs) in the inner leaflet, plays a key role in antibiotic ...The outer membrane (OM) of Gram-negative bacteria, which consists of lipopolysaccharides (LPS) in the outer leaflet and phospholipids (PLs) in the inner leaflet, plays a key role in antibiotic resistance and pathogen virulence. The maintenance of lipid asymmetry (Mla) pathway is known to be involved in PL transport and contributes to the lipid homeostasis of the OM, yet the underlying molecular mechanism and the directionality of PL transport in this pathway remain elusive. Here, we reported the cryo-EM structures of the ATP-binding cassette (ABC) transporter MlaFEBD from P. areuginosa, the core complex in the Mla pathway, in nucleotide-free (apo)-, ADP (ATP + vanadate)- and ATP (AMPPNP)-bound states as well as the structures of MlaFEB from E. coli in apo- and AMPPNP-bound states at a resolution range of 3.4-3.9 Å. The structures show that the MlaFEBD complex contains a total of twelve protein molecules with a stoichiometry of MlaFEBD, and binds a plethora of PLs at different locations. In contrast to canonical ABC transporters, nucleotide binding fails to trigger significant conformational changes of both MlaFEBD and MlaFEB in the nucleotide-binding and transmembrane domains of the ABC transporter, correlated with their low ATPase activities exhibited in both detergent micelles and lipid nanodiscs. Intriguingly, PLs or detergents appeared to relocate to the membrane-proximal end from the distal end of the hydrophobic tunnel formed by the MlaD hexamer in MlaFEBD upon addition of ATP, indicating that retrograde PL transport might occur in the tunnel in an ATP-dependent manner. Site-specific photocrosslinking experiment confirms that the substrate-binding pocket in the dimeric MlaE and the MlaD hexamer are able to bind PLs in vitro, in line with the notion that MlaFEBD complex functions as a PL transporter. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 59.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 15.4 KB 15.4 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 9.2 KB | Display | ![]() |
Images | ![]() | 57.7 KB | ||
Filedesc metadata | ![]() | 5.8 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7ch9MC ![]() 7ch6C ![]() 7ch7C ![]() 7ch8C ![]() 7chaC M: atomic model generated by this map C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Voxel size | X=Y=Z: 1 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Cryo-EM structure of P.aeruginosa MlaFEBD
Entire | Name: Cryo-EM structure of P.aeruginosa MlaFEBD |
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Components |
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-Supramolecule #1: Cryo-EM structure of P.aeruginosa MlaFEBD
Supramolecule | Name: Cryo-EM structure of P.aeruginosa MlaFEBD / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#4 |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: MlaD domain-containing protein
Macromolecule | Name: MlaD domain-containing protein / type: protein_or_peptide / ID: 1 / Number of copies: 6 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() Strain: ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1 |
Molecular weight | Theoretical: 16.571945 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MQTRTLEIGV GLFLLAGLLA LLLLALRVSG LSVGNAGDTY KVYAYFDNIA GVTVRGKVTL AGVTIGKVTA VDLDRDSYTG RVTMEINQN VNNLPVDSTA SILTAGLLGE KYIGISVGGD EDVLKDGSTI HDTQSALVLE DLIGKFLLNS VNKDEAKK UniProtKB: Mce/MlaD domain-containing protein |
-Macromolecule #2: Probable permease of ABC transporter
Macromolecule | Name: Probable permease of ABC transporter / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() Strain: ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1 |
Molecular weight | Theoretical: 28.400326 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MRRVSPLERI RLFGRAGLDV VAALGRSTLF LGHALLGRRT PGTGLHLLVK QLYSVGVLSL AIIVVSGLFI GMVLALQGYN ILISYGSEQ AVGQMVALTL LRELGPVVTG LLFAGRAGSA LTAEIGNMKA TEQLSSLEMI GVDPLKYIVA PRLWAGFISM P LLAAIFSV ...String: MRRVSPLERI RLFGRAGLDV VAALGRSTLF LGHALLGRRT PGTGLHLLVK QLYSVGVLSL AIIVVSGLFI GMVLALQGYN ILISYGSEQ AVGQMVALTL LRELGPVVTG LLFAGRAGSA LTAEIGNMKA TEQLSSLEMI GVDPLKYIVA PRLWAGFISM P LLAAIFSV VGIWGGAMVA VDWLGVYEGS FWANMQNSVQ FTEDVLNGVI KSIVFAFVVT WIAVYQGYDC EPTSEGISRA TT RTVVYAS LAVLGLDFIL TALMFGDF UniProtKB: Intermembrane phospholipid transport system permease protein MlaE |
-Macromolecule #3: Probable ATP-binding component of ABC transporter
Macromolecule | Name: Probable ATP-binding component of ABC transporter / type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() Strain: ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1 |
Molecular weight | Theoretical: 29.657367 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MSTDSAYAVE LKGLTFKRGS RAIFDNIDVR IPRGKVTGIM GPSGCGKTTL LRLIASQLRP SKGEVWVNGQ NLPQLSRGDL FDMRKQFGV LFQSGALFTD LDVFENVAFP LRVHTQLPEE MIRDIVLMKL QAVGLRGAVE LMPDELSGGM KRRVALARAI A LDPQILLY ...String: MSTDSAYAVE LKGLTFKRGS RAIFDNIDVR IPRGKVTGIM GPSGCGKTTL LRLIASQLRP SKGEVWVNGQ NLPQLSRGDL FDMRKQFGV LFQSGALFTD LDVFENVAFP LRVHTQLPEE MIRDIVLMKL QAVGLRGAVE LMPDELSGGM KRRVALARAI A LDPQILLY DEPFVGQDPI AMGVLVRLIR LLNDALGITS IVVSHDLAET ASIADYIYIV GDGRVLGHGT PDVLKETDDP RI RQFVKGI PDGPVPFHYP ARDYRADLLG ER UniProtKB: Probable ATP-binding component of ABC transporter |
-Macromolecule #4: STAS domain-containing protein
Macromolecule | Name: STAS domain-containing protein / type: protein_or_peptide / ID: 4 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() Strain: ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1 |
Molecular weight | Theoretical: 10.789388 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MSQASLREGA AGELQLAGVL DYSSGPALRE QGGRLIRASQ AAELVVDCSA VERSSSVGIS LLLAFIRDAR KAGKVLSVRA LPDDMREIA KVSSLLEILP LQE UniProtKB: STAS domain-containing protein |
-Macromolecule #5: 2-(HEXADECANOYLOXY)-1-[(PHOSPHONOOXY)METHYL]ETHYL HEXADECANOATE
Macromolecule | Name: 2-(HEXADECANOYLOXY)-1-[(PHOSPHONOOXY)METHYL]ETHYL HEXADECANOATE type: ligand / ID: 5 / Number of copies: 10 / Formula: LPP |
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Molecular weight | Theoretical: 648.891 Da |
Chemical component information | ![]() ChemComp-LPP: |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 8 |
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Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Average electron dose: 60.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: TUNGSTEN HAIRPIN |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: SPOT SCAN / Imaging mode: DIFFRACTION / Cs: 2.7 mm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |