|Entry||Database: PDB / ID: 6aye|
|Title||Human apo-TRPML3 channel at pH 7.4|
|Keywords||TRANSPORT PROTEIN / ion channel / TRP channel / lysosomal|
|Specimen source||Homo sapiens / human|
|Method||Electron microscopy (4.06 Å resolution / Particle / Single particle)|
|Authors||Zhou, X. / Li, M. / Su, D. / Jia, Q. / Li, H. / Li, X. / Yang, J.|
|Citation||Nat. Struct. Mol. Biol., 2017, 24, 1146-1154|
SummaryFull reportAbout validation report
|Date||Deposition: Sep 8, 2017 / Release: Nov 8, 2017|
Downloads & links
Mass: 64625.785 Da / Num. of mol.: 4 / Source: (gene. exp.) Homo sapiens / human / References: UniProt: Q8TDD5
|Experiment||Method: ELECTRON MICROSCOPY|
|EM experiment||Aggregation state: PARTICLE / Reconstruction method: SINGLE PARTICLE|
|Component||Name: human TRPML3 apo channel at pH 7.4 / Type: COMPLEX / Entity ID: 1 / Source: RECOMBINANT|
|Molecular weight||Experimental value: NO|
|Source (natural)||Organism: Homo sapiens|
|Source (recombinant)||Organism: Trichoplusia ni|
|Buffer solution||Details: The pH of the buffer was adjusted to 7.4 by NaOH. / pH: 7.4|
|Specimen||Conc.: 1.9 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES|
|Specimen support||Grid material: COPPER / Grid mesh size: 400 / Grid type: Quantifoil holey carbon grid R1.2/1.3|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 281 kelvins|
Details: waiting for 3 seconds before blotting for 4 seconds(double-sided, blot force 1),then the grid was immediately plunged into liquid ethane cooled by liquid-nitrogen
-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: SPOT SCAN|
|Electron lens||Mode: BRIGHT FIELD / Nominal magnification: 22500 / Nominal defocus max: 3000 nm / Nominal defocus min: 1500 nm / Cs: 2.7 mm / Alignment procedure: COMA FREE|
|Specimen holder||Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER|
|Image recording||Average exposure time: 8 sec. / Electron dose: 50 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K2 SUMMIT (4k x 4k)|
|Image scans||Sampling size: 5 microns / Dimension width: 7676 / Dimension height: 7420 / Movie frames/image: 32 / Used frames/image: 1-32|
|CTF correction||Type: NONE|
|Symmetry||Point symmetry: C4|
|3D reconstruction||Resolution: 4.06 Å / Resolution method: FSC 0.143 CUT-OFF / Number of particles: 43542 / Symmetry type: POINT|
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