3.2 Å Single-Particle Cryo-EM Reconstruction of 49 kDa Membrane-Bound PfCRT Complexed with Fab

REL 2019-10-06 2019-12-19 2020-01-29 3.2 Å Single-Particle Cryo-EM Reconstruction of 49 kDa Membrane-Bound PfCRT Complexed with Fab 0000-0001-6656-6320 Yong Zi Tan New York Structural Biology Center/Columbia University 89 Convent Ave New York New York United States 10027 0000-0003-3293-2200 Filippo Mancia Department of Physiology and Cellular Biophysics, Columbia University Irving Medical Center New York New York United States NY 10032 0000-0001-6753-8938 David Armand Fidock Division of Infectious Diseases, Department of Medicine, Columbia University Irving Medical Center New York New York United States NY 10032 0000-0002-8620-827X Matthias Quick Department of Psychiatry, Center for Molecular Recognition, Columbia University Irving Medical Center New York New York United States NY 10032 Kim JK Tan YZT Wicht KJW Erramilli SKE Dhingra SKD Okombo JO Vendome JV Hagenah LMH Giacometti SIG Warren ALW Nosol KN Roepe PDR Potter CSP Carragher BC Kossiakoff AAK Quick MQ Fidock DAF Mancia FM 830.4 10.6019/EMPIAR-10330 EMDB EMD-20806 Kim JK Tan YZT Wicht KJW Erramilli SKE Dhingra SKD Okombo JO Vendome JV Hagenah LMH Giacometti SIG Warren ALW Nosol KN Roepe PDR Potter CSP Carragher BC Kossiakoff AAK Quick MQ Fidock DAF Mancia FM Structure and drug resistance of the Plasmodium falciparum transporter PfCRT Nature Nature United Kingdom 7786 576 315 320 2019 English 10.1038/s41586-019-1795-x 31776516 Unaligned and compressed multi-frame micrographs /data/01_Raw_Frames micrographs - multiframe MRCS MRCS 3383 80 1 80 UNSIGNED 16 BIT INTEGER 3838 0.5175 3710 0.5175

Data collection was performed using a dose of ~91.56 e-/Å2 across 80 frames (75 msec per frame) at a dose rate of ~4.0 e–/pix/sec, using a set defocus range of -1.2 μm to -1.8 μm. Raw frames were compressed using pbzip2. Gain reference image to use is 18jun08i_08213559_01_3710x3838_norm_0_mod.mrc. Magnification anisotropy is present (Distortion angle of 172.7 degrees, major scale of 1.007 and minor scale of 0.993). The script to correct for magnification anisotropy (mag_distort_correct4.cs is attached, and it requires the mag_distortion_correct software available from the Grigorieff lab website.

Aligned and magnification anisotropy corrected micrographs /data/02_Aligned_Micrographs micrographs - single frame MRC MRC 3297 1 32 BIT FLOAT 3838 0.5175 3710 0.5175

Micrographs aligned using MotionCor2, magnification anisotropy corrected. Alignment was done initially using 3x3 patches (Patch3_micrographs). Micrographs that had estimated resolution at 0.5 confidence level of worse than 10 Å were removed, resulting in 3,297 micrographs. Patch2_micrographs were used later during refinement - hence only a subset of micrographs were aligned using 2x2 patches (Patch2_micrographs).

Final Particle Stack with Refined Euler Angles and Shifts /data/03_Final_Refined_Particle_Stack picked particles - single frame - processed MRC MRC 16905 1 32 BIT FLOAT 300 1.035 300 1.035

Final particle stack, with Relion .star file containing the refined Euler angles and shifts.

data/02_Aligned_Micrographs/Patch*_micrographs/*mrc data/03_Final_Refined_Particle_Stack/FinalRefinement-OriginalParticles-PfCRT.star data/03_Final_Refined_Particle_Stack