REL 2021-01-18 2021-04-14 2021-06-30 0000-0001-8102-9769 Natalya Leneva Cambridge Institute for Medical Research Hills Rd Cambridge United Kingdom CB2 0XY 0000-0003-3990-6910 John A G Briggs MRC LMB Francis Crick Avenue Cambridge United Kingdom CB2 0QH 0000-0002-8351-6322 David J Owen Cambridge Institute for Medical Research Cambridge Biomedical Campus Hills Road Cambridge United Kingdom CB2 0XY Leneva N Kovtun O Morado DR Briggs JAG Owen DJ 764.9 10.6019/EMPIAR-10633 EMDB EMD-12220 EMD-12221 EMD-12222 Leneva N Kovtun O Morado DR Briggs JAG Owen DJ Science advances Sci Adv United Kingdom 13 7 2021 English 10.1126/sciadv.abf8598 33762348

Raw frames, motion-corrected tilt series, defocus estimation and IMOD tomogram reconstruction files for the membrane-reconstituted metazoan retromer:SXN3 complex in the presence of cargo-signal containing C-portion of the Wls receptor. Collected on Titan Krios at 300 kV using energy-filtered (20eV slit) Gatan K3 camera. Dose-symmetric tilt series collection was done using SerialEM 3.8.0 (Tilt Series Controler), 3 degrees tilt step, +/- 60 degrees range. Ten frames of movies were recorded for each tilt. The exposure dose per tilt image was 3.0 e/Å^2. For gain correction, use data/correction/SuperRef_ts_01_14.dm4 for tilts ts_01 to 14, and data/correction/SuperRef_ts_16_126.dm4 for tilts ts_16 to 126. The defect file data/correction/Krios3_K3_SerialEM_defects.txt to be applied to all tilt series.

Raw image frames for the metazoan retromer:SNX3 coat assembled on the Wls cargo-containing membranes /data/rawframes micrographs - multiframe MRC TIFF 4844 10 1 10 UNSIGNED BYTE 11520 0.8505 8184 0.8505

Unaligned superresolution frames not corrected for detector’s gain and defects. Ten frames were saved per each tilt image. The exposure dose per tilt image was 3.0 e/Å^2. Calibrated physical pixel size 1.701 Å. Movies are named as ts_[tilt stack ID]_[sequntial movie number]_[tilt angle].tif. For gain reference, use data/correction/SuperRef_ts_01_14.dm4 for tilts ts_01 to 14, and data/correction/SuperRef_ts_16_126.dm4 for tilts ts_16 to 126. The defect file data/correction/Krios3_K3_SerialEM_defects.txt to be applied to all tilt series.

Corrected, aligned and order-sorted tilt series for the metazoan retromer:SNX3 coat assembled on the Wls cargo-containing membranes /data/aligned_stacks tilt series MRC MRC 113 41 1 41 SIGNED 16 BIT INTEGER 5760 1.701 4092 1.701

Corrected, aligned and order-sorted tilt series. The tilt-series were generated from raw frames (data/rawframes) by applying gain reference and defect corrections, motion correction and Fourier cropping to physical pixel size. No dose-filtering was applied. Defocus estimation was performed using IMOD ctfplotter on non-dose filtered aligned stacks (data/aligned_stacks). ETOMO generated tilt alignment using 10 nm gold fiducials tracking.

Corrected, aligned, dose-filtered and order-sorted tilt series for the metazoan retromer:SNX3 coat assembled on the Wls cargo-containing membranes /data/aligned_dose_filt_stacks tilt series MRC MRC 113 41 1 41 SIGNED 16 BIT INTEGER 5760 1.701 4092 1.701

Dose-filtered aligned stacks.