2021-04-14 REL PDBe PDBe PDBe 2020-03-12 2020-07-29 2020-07-29 2021-04-14 Wellcome Trust 207455/Z/17/Z United Kingdom Medical Research Council (MRC, United Kingdom) MC_UP_1201/16 United Kingdom Pentagon bounding clathrin legs in clathrin coats assembled on a membrane Kovtun O Kane Dickson V Kelly BT Owen D Briggs JAG Kovtun O Dickson VK Kelly BT Owen DJ Briggs JAG Architecture of the AP2/clathrin coat on the membranes of clathrin-coated vesicles. Sci Adv US 6 eaba8381 eaba8381 2020 32743075 doi:10.1126/sciadv.aba8381 2375-2548 EMD-10747 other EM volume
AP2 core in physiological buffer
 EMD-10748 other EM volume EMD-10749 other EM volume EMD-10750 other EM volume EMD-10751 other EM volume EMD-10752 other EM volume EMD-10753 associated EM volume EMD-10754 other EM volume Clathrin/AP2 coat assembled on membrane. Clathrin/AP2 coat assembled on membrane. 0
Clathrin/AP coat was formed on the membrane containing cargo signal peptides. The AP2 adaptor lacked hinge and appendage regions in its alpha subunit.
 Natural Sus scrofa component 1 Sus scrofa Recombinant Homo sapiens component 1 Homo sapiens Escherichia coli 'BL21-Gold(DE3)pLysS AG' subtomogramAveraging threeDArray 0.7 7.2 10.0 Hepes KOH 120.0 Potassium Acetate 2.0 Magnesium chloride 5.0 2-Mercaptoethanol C-flat-2/2 CARBON HOLEY ETHANE 98 291 LEICA EM GP
The sample was supplemented with 10 nm nanogold fiducials, and 3 ul of the mixture was backside blotted for 3 seconds..
The sample (in vitro budding reaction) contained AP2, clathrin and 400 nm extruded liposomes
 FEI TITAN KRIOS FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 300 100.0 2.7 1.5 1.5 6.5 6.5 81000.0 81000.0 FEI TITAN KRIOS AUTOGRID HOLDER NITROGEN GIF Quantum LS 20 GATAN K2 QUANTUM (4k x 4k) COUNTING 3838 3710 1-10 2 0.2 3.2
The images were collected in movie mode at 10 frames per second
 1
The images were low pass filtered according to the cumulative radiation dose.
C1 7.7 FSC 0.143 CUT-OFF TOM AV3 24759 58121 198871 reference-free geometrically defined initial positions TOM
To define initial positions of subtomograms, centres and radii of coated vesicles, buds were manually marked in bin4 tomograms. These measurements were used to define spheres. Subtomogram positions were then defined on the surface of these spheres with uniform sampling. The orientations were calculated to be normal to the surfaces of the spheres with random in-plane rotation.
 NOVACTF 1.0
CTF correction in novaCTF with by multiplication
 PEET Dynamo OTHER TOM AV3 emd_10753.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 160 160 160 0 0 0 160 160 160 284.8 284.8 284.8 90.0 90.0 90.0 X Y Z -2.5666678 4.6766224 0.0010469395 0.30047828 1.78 1.78 1.78 1.5 AUTHOR Pentagon bounding clathrin leg 1XI4 6SCT 1E42 A FLEXIBLE FIT correlation coefficient REAL emd_10753_msk_1.map emd_10753_half_map_1.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 160 160 160 0 0 0 160 160 160 284.8 284.8 284.8 90.0 90.0 90.0 X Y Z -1.2419084 1.5605103 -0.0003843486 0.18296869 1.78 1.78 1.78 AUTHOR emd_10753_half_map_2.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 160 160 160 0 0 0 160 160 160 284.8 284.8 284.8 90.0 90.0 90.0 X Y Z -1.2297754 1.6203508 -0.00030315507 0.1754459 1.78 1.78 1.78 AUTHOR