[English] 日本語
![](img/lk-miru.gif)
- EMDB-6025: Conformational Spectrum of Multidrug ABC Transporters Revealed by... -
+
Open data
-
Basic information
Entry | Database: EMDB / ID: EMD-6025 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Conformational Spectrum of Multidrug ABC Transporters Revealed by Single Particle Electron Microscopy | |||||||||
![]() | MsbA in inward-facing conformation | |||||||||
![]() |
| |||||||||
![]() | ABC transporter / P-gp / MsbA | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 30.0 Å | |||||||||
![]() | Moeller A / Chang Lee S / Tao H / Speir S / Chang G / Urbatsch IL / Potter CS / Carragher B / Zhang Q | |||||||||
![]() | ![]() Title: Distinct conformational spectrum of homologous multidrug ABC transporters. Authors: Arne Moeller / Sung Chang Lee / Houchao Tao / Jeffrey A Speir / Geoffrey Chang / Ina L Urbatsch / Clinton S Potter / Bridget Carragher / Qinghai Zhang / ![]() Abstract: ATP-binding cassette (ABC) exporters are ubiquitously found in all kingdoms of life and their members play significant roles in mediating drug pharmacokinetics and multidrug resistance in the clinic. ...ATP-binding cassette (ABC) exporters are ubiquitously found in all kingdoms of life and their members play significant roles in mediating drug pharmacokinetics and multidrug resistance in the clinic. Significant questions and controversies remain regarding the relevance of their conformations observed in X-ray structures, their structural dynamics, and mechanism of transport. Here, we used single particle electron microscopy (EM) to delineate the entire conformational spectrum of two homologous ABC exporters (bacterial MsbA and mammalian P-glycoprotein) and the influence of nucleotide and substrate binding. Newly developed amphiphiles in complex with lipids that support high protein stability and activity enabled EM visualization of individual complexes in a membrane-mimicking environment. The data provide a comprehensive view of the conformational flexibility of these ABC exporters under various states and demonstrate not only similarities but striking differences between their mechanistic and energetic regulation of conformational changes. | |||||||||
History |
|
-
Structure visualization
Movie |
![]() |
---|---|
Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
-
Downloads & links
-EMDB archive
Map data | ![]() | 134.7 KB | ![]() | |
---|---|---|---|---|
Header (meta data) | ![]() ![]() | 9.1 KB 9.1 KB | Display Display | ![]() |
Images | ![]() | 202.4 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 78.7 KB | Display | ![]() |
---|---|---|---|---|
Full document | ![]() | 77.7 KB | Display | |
Data in XML | ![]() | 495 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6007C ![]() 6018C ![]() 6019C ![]() 6020C ![]() 6021C ![]() 6022C ![]() 6023C ![]() 6024C ![]() 6026C ![]() 6027C ![]() 6028C ![]() 6029C ![]() 6030C ![]() 6031C ![]() 6032C ![]() 6033C C: citing same article ( |
---|---|
Similar structure data |
-
Links
EMDB pages | ![]() ![]() |
---|
-
Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | MsbA in inward-facing conformation | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. generated in cubic-lattice coordinate | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 5 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-
Sample components
-Entire : MsbA stabilised in amphiphilic environment - inward-facing 4.5 nm...
Entire | Name: MsbA stabilised in amphiphilic environment - inward-facing 4.5 nm opening |
---|---|
Components |
|
-Supramolecule #1000: MsbA stabilised in amphiphilic environment - inward-facing 4.5 nm...
Supramolecule | Name: MsbA stabilised in amphiphilic environment - inward-facing 4.5 nm opening type: sample / ID: 1000 / Oligomeric state: homodimer / Number unique components: 1 |
---|---|
Molecular weight | Theoretical: 140 KDa |
-Macromolecule #1: MsbA
Macromolecule | Name: MsbA / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Oligomeric state: dimer / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 140 KDa |
Recombinant expression | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | negative staining |
---|---|
![]() | single particle reconstruction |
Aggregation state | particle |
-
Sample preparation
Staining | Type: NEGATIVE / Details: uranyl formate |
---|---|
Grid | Details: thin carbon over holes |
Vitrification | Cryogen name: NONE / Instrument: OTHER |
-
Electron microscopy
Microscope | FEI TECNAI F20 |
---|---|
Date | Jan 24, 2012 |
Image recording | Category: CCD / Film or detector model: TVIPS TEMCAM-F416 (4k x 4k) / Number real images: 558 / Average electron dose: 45 e/Å2 |
Tilt angle min | 0 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.75 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 64000 |
Sample stage | Specimen holder model: SIDE ENTRY, EUCENTRIC / Tilt angle max: 55 |
Experimental equipment | ![]() Model: Tecnai F20 / Image courtesy: FEI Company |
-
Image processing
Details | RCT reconstruction |
---|---|
CTF correction | Details: whole micrograph - unitilted images |
Final reconstruction | Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 30.0 Å / Resolution method: OTHER / Software - Name: Appion, Spider, Sparx, Eman2 / Number images used: 148 |