REL 2023-11-16 2025-07-03 2025-07-03 0000-0002-1332-128X Elizabeth Campbell Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 1230 York Avenue New York New York United States 10065 0000-0002-1332-128X Elizabeth Campbell Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 1230 York Avenue New York New York United States 10065 0000-0002-8241-3153 Seth Darst Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 1230 York Avenue New York New York United States 10065 Campbell EA Small GI Darst SA National Institutes of Health (NIH) AI161278 United States 5.3 10.6019/EMPIAR-11803 EMDB molecule EMD-40699 Small GI Fedorova O Olinares PDB Chandanani J Banerjee A Choi YJ Molina H Chait BT Darst SA Campbell EA Molecular cell Mol Cell United States 21 83 3921 3930.e7 2023 English 10.1016/j.molcel.2023.10.001 37890482

Grids were imaged using a 300 kV Titan Krios (Thermo Fisher Scientific) equipped with a GIF BioQuantum and K3 camera (Gatan). Movies were collected with Leginon60 with a pixel size of 1.0825 Å per px (micrograph dimensions of 5,760 × 4,092 px) over a defocus range of 0.7 to 2.5 μm with a 20 eV energy filter slit. Videos were recorded in counting mode (native K3 camera binning 2) with ∼25 e−/ Å2/s exposure rate in dose-fractionation mode with intermediate frames recorded every 50 ms over a 2 s exposure (40 frames per micrograph) to give an electron exposure of ∼51 e−/Å2. Dose-fractionated videos were gain-normalized, drift-corrected, summed and dose-weighted using MotionCor2.48 The CTF was estimated for each summed image using the Patch CTF module in cryoSPARC v.3.2.0.46 Particles were picked and extracted from the dose-weighted images with box size of 256 px using cryoSPARC blob picker and particle extraction. The entire dataset consisted of 21,859 motion-corrected images with 10,257,549 particles.

Unaligned multiframe micrographs /data/rawdata micrographs - multiframe TIFF TIFF 22411 40 UNSIGNED BYTE variable variable variable variable

Multiframe micrographs in TIFF format that were used as the basis for processing and determining a cryo-EM map for the UMP-i structure (EMD-40699, PDB-8SQ9).