Protein or peptide: 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma
Ligand: CALCIUM ION
Keywords
1-phosphatidylinositol 4 / 5-bisphosphate phosphodiesterase gamma-1 cryo EM / HYDROLASE
Function / homology
Function and homology information
phosphoinositide phospholipase C / phosphatidylinositol-4,5-bisphosphate phospholipase C activity / phospholipid catabolic process / intracellular signal transduction / calcium ion binding Similarity search - Function
Name: CALCIUM ION / type: ligand / ID: 2 / Number of copies: 1 / Formula: CA
Molecular weight
Theoretical: 40.078 Da
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Experimental details
-
Structure determination
Method
cryo EM
Processing
single particle reconstruction
Aggregation state
particle
-
Sample preparation
Concentration
10 mg/mL
Buffer
pH: 7.4 Component:
Concentration
Formula
Name
20.0 mM
C8H18N2O4S
HEPES
100.0 mM
NaCl
NaCl
10.0 mM
MgCl2
MgCl2
2.0 mM
C4H10O2S2
DTT
0.15 mM
C24H46O11
n-Dodecyl-B-D-Maltoside
Grid
Model: C-flat-1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Support film - Film thickness: 12 / Pretreatment - Type: PLASMA CLEANING
Vitrification
Cryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 295 K / Instrument: FEI VITROBOT MARK IV
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Electron microscopy
Microscope
FEI TALOS ARCTICA
Image recording
Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Digitization - Dimensions - Width: 5760 pixel / Digitization - Dimensions - Height: 4092 pixel / Number grids imaged: 1 / Number real images: 4450 / Average electron dose: 55.0 e/Å2
Electron beam
Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron optics
Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm
Sample stage
Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company
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Image processing
Startup model
Type of model: NONE
Final reconstruction
Number classes used: 13 / Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.68 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 3.2) / Number images used: 238458
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