|Entry||Database: PDB / ID: 6e0h|
|Title||PDB: afTMEM16 reconstituted in nanodiscs in the presence of Ca2+|
|Components||Plasma membrane channel protein (Aqy1), putative|
|Keywords||LIPID TRANSPORT / scramblase / Ca2+-activated / membrane-reorganization|
|Function / homology||Calcium-activated chloride channel / Anoctamin / phospholipid scramblase activity / phospholipid translocation / voltage-gated calcium channel activity / ion transmembrane transport / integral component of membrane / Plasma membrane channel protein (Aqy1), putative|
Function and homology information
|Specimen source||Neosartorya fumigata (mold)|
|Method||ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / 4.05 Å resolution|
|Authors||Falzone, M.E. / Accardi, A.|
|Citation||Journal: Elife / Year: 2019|
Title: Structural basis of Ca-dependent activation and lipid transport by a TMEM16 scramblase.
Authors: Maria E Falzone / Jan Rheinberger / Byoung-Cheol Lee / Thasin Peyear / Linda Sasset / Ashleigh M Raczkowski / Edward T Eng / Annarita Di Lorenzo / Olaf S Andersen / Crina M Nimigean / Alessio Accardi
Abstract: The lipid distribution of plasma membranes of eukaryotic cells is asymmetric and phospholipid scramblases disrupt this asymmetry by mediating the rapid, nonselective transport of lipids down their ...The lipid distribution of plasma membranes of eukaryotic cells is asymmetric and phospholipid scramblases disrupt this asymmetry by mediating the rapid, nonselective transport of lipids down their concentration gradients. As a result, phosphatidylserine is exposed to the outer leaflet of membrane, an important step in extracellular signaling networks controlling processes such as apoptosis, blood coagulation, membrane fusion and repair. Several TMEM16 family members have been identified as Ca-activated scramblases, but the mechanisms underlying their Ca-dependent gating and their effects on the surrounding lipid bilayer remain poorly understood. Here, we describe three high-resolution cryo-electron microscopy structures of a fungal scramblase from , afTMEM16, reconstituted in lipid nanodiscs. These structures reveal that Ca-dependent activation of the scramblase entails global rearrangement of the transmembrane and cytosolic domains. These structures, together with functional experiments, suggest that activation of the protein thins the membrane near the transport pathway to facilitate rapid transbilayer lipid movement.
SummaryFull reportAbout validation report
|Date||Deposition: Jul 6, 2018 / Release: Feb 6, 2019|
|Structure viewer||Molecule: |
Downloads & links
B: Plasma membrane channel protein (Aqy1), putative
A: Plasma membrane channel protein (Aqy1), putative
Mass: 84616.859 Da / Num. of mol.: 2
Source: (gene. exp.) Neosartorya fumigata (strain ATCC MYA-4609 / Af293 / CBS 101355 / FGSC A1100) (mold)
Strain: ATCC MYA-4609 / Af293 / CBS 101355 / FGSC A1100 / Gene: AFUA_4G02970 / Production host: Saccharomyces cerevisiae (baker's yeast) / References: UniProt: Q4WA18
|Experiment||Method: ELECTRON MICROSCOPY|
|EM experiment||Aggregation state: PARTICLE / Reconstruction method: single particle reconstruction|
|Component||Name: afTMEM16 reconstituted in nanodiscs in the presence of Ca2+|
Type: COMPLEX / Entity ID: 1 / Source: RECOMBINANT
|Molecular weight||Value: 168 kDa/nm / Experimental value: NO|
|Source (natural)||Organism: Aspergillus fumigatus (mold)|
|Source (recombinant)||Organism: Saccharomyces cerevisiae (baker's yeast)|
|Buffer solution||pH: 8|
|Specimen||Conc.: 7 mg/ml|
Details: afTMEM16 reconstituted in nanodiscs in the presence of Ca2+
Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 288 kelvins|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Microscopy||Microscope model: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: OTHER|
|Electron lens||Mode: BRIGHT FIELDBright-field microscopy|
|Image recording||Electron dose: 69.97 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Number of grids imaged: 1|
|CTF correction||Type: PHASE FLIPPING AND AMPLITUDE CORRECTION|
|Symmetry||Point symmetry: C2|
|3D reconstruction||Resolution: 4.05 Å / Resolution method: FSC 0.143 CUT-OFF / Number of particles: 37146 / Symmetry type: POINT|
|Refine||Stereochemistry target values: GeoStd + Monomer Library|
|Number of atoms included #1||Protein: 9520 / Nucleic acid: 0 / Ligand: 4 / Solvent: 0|
|Refine LS restraints|
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