|Entry||Database: EMDB / ID: 8669|
|Title||CryoEM map of MsbA in POPG nanodisc|
|Map data||map of MsbA with LPS in POPG nanodisc, filtered to 5A and sharpened with -280 B-factor|
|Sample||MsbA with LPS in POPG nanodisc:|
|Function / homology||ABC transporter, conserved site / ABC transporters family signature. / Lipid A export ATP-binding/permease protein msbA family profile. / ABC transporter integral membrane type-1 fused domain profile. / ABC transporter-like / AAA+ ATPase domain / ABC transporter type 1, transmembrane domain / ABC transporter, lipid A export, MsbA / ATP-binding cassette, ABC transporter-type domain profile. / P-loop containing nucleoside triphosphate hydrolase ...ABC transporter, conserved site / ABC transporters family signature. / Lipid A export ATP-binding/permease protein msbA family profile. / ABC transporter integral membrane type-1 fused domain profile. / ABC transporter-like / AAA+ ATPase domain / ABC transporter type 1, transmembrane domain / ABC transporter, lipid A export, MsbA / ATP-binding cassette, ABC transporter-type domain profile. / P-loop containing nucleoside triphosphate hydrolase / ABC transporter type 1, transmembrane domain superfamily / ABC transporter / ABC transporter transmembrane region / lipid-transporting ATPase activity / ec:3.6.3.-: / integral component of membrane / ATP binding / plasma membrane / Lipid A export ATP-binding/permease protein MsbA|
Function and homology information
|Source||Escherichia coli (E. coli)|
|Method||single particle reconstruction / cryo EM / 5 Å resolution|
|Authors||Mi W / Walz T / Liao M|
|Citation||Journal: Nature / Year: 2017|
Title: Structural basis of MsbA-mediated lipopolysaccharide transport.
Authors: Wei Mi / Yanyan Li / Sung Hwan Yoon / Robert K Ernst / Thomas Walz / Maofu Liao
Abstract: Lipopolysaccharide (LPS) in the outer membrane of Gram-negative bacteria is critical for the assembly of their cell envelopes. LPS synthesized in the cytoplasmic leaflet of the inner membrane is ...Lipopolysaccharide (LPS) in the outer membrane of Gram-negative bacteria is critical for the assembly of their cell envelopes. LPS synthesized in the cytoplasmic leaflet of the inner membrane is flipped to the periplasmic leaflet by MsbA, an ATP-binding cassette transporter. Despite substantial efforts, the structural mechanisms underlying MsbA-driven LPS flipping remain elusive. Here we use single-particle cryo-electron microscopy to elucidate the structures of lipid-nanodisc-embedded MsbA in three functional states. The 4.2 Å-resolution structure of the transmembrane domains of nucleotide-free MsbA reveals that LPS binds deep inside MsbA at the height of the periplasmic leaflet, establishing extensive hydrophilic and hydrophobic interactions with MsbA. Two sub-nanometre-resolution structures of MsbA with ADP-vanadate and ADP reveal an unprecedented closed and an inward-facing conformation, respectively. Our study uncovers the structural basis for LPS recognition, delineates the conformational transitions of MsbA to flip LPS, and paves the way for structural characterization of other lipid flippases.
|Date||Deposition: Apr 11, 2017 / Header (metadata) release: Jul 19, 2017 / Map release: Sep 20, 2017 / Last update: Sep 27, 2017|
|Structure viewer||EM map: |
Downloads & links
|File||emd_8669.map.gz (map file in CCP4 format, 28312 KB)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.28 Å|
CCP4 map header:
-Entire MsbA with LPS in POPG nanodisc
|Entire||Name: MsbA with LPS in POPG nanodisc / Number of components: 2|
|Mass||Theoretical: 67.2 kDa|
-Component #1: protein, MsbA with LPS in POPG nanodisc
|Protein||Name: MsbA with LPS in POPG nanodisc / Recombinant expression: No|
|Mass||Theoretical: 67.2 kDa|
|Source||Species: Escherichia coli (E. coli)|
|Source (engineered)||Expression System: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (bacteria)|
-Component #2: protein, MsbA
|Protein||Name: MsbA / Recombinant expression: No|
|Source (engineered)||Expression System: Escherichia coli (E. coli)|
|Specimen||Specimen state: particle / Method: cryo EM|
|Sample solution||Specimen conc.: 1 mg/ml / pH: 7.5|
|Vitrification||Cryogen name: ETHANE|
-Electron microscopy imaging
Model: Tecnai Polara / Image courtesy: FEI Company
|Imaging||Microscope: FEI POLARA 300|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 47 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 (4k x 4k)|
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