REL 2023-08-01 2024-07-09 2024-07-09 0000-0001-8667-6797 Judy Hirst MRC mitochondrial Biology Unit, University of Cambridge The Keith Peters Building, Cambridge Biomedical Campus, Hills Road Cambridge Cambridgeshire United Kingdom CB2 0XY 0000-0001-8667-6797 Judy Hirst MRC mitochondrial Biology Unit, University of Cambridge The Keith Peters Building, Cambridge Biomedical Campus, Hills Road Cambridge Cambridgeshire United Kingdom CB2 0XY Grba DN Wright JJ Fisher W Yin Z Hirst J Medical Research Council (MRC, United Kingdom) MC_UU_00015/2 United Kingdom Medical Research Council (MRC, United Kingdom) MC_UU_00028/1 United Kingdom 1.4 10.6019/EMPIAR-11637 EMDB molecule EMD-18066 EMD-18067 EMD-18068 EMD-18069 Grba DN Wright JJ Fisher W Yin Z Hirst J Science (New York, N.Y.) Science 6701 384 1247 1253 2024 English 10.1126/science.ado2075 38870289 EMPIAR-11636 Unaligned multi-frame micrographs of wild-type bos taurus mitochondrial complex I, solubilised in LMNG, reconstituted into proteoliposomes and deactivated. Non-tilted collection. /data micrographs - multiframe TIFF TIFF 6692 40 1 40 UNSIGNED BYTE 5760 1.072 4092 1.072

Unaligned multi-frame micrographs of wild-type bovine mitochondrial complex I from an as-prepared sample solubilised in LMNG and reconstituted in to liposomes. Not gain corrected (gain file included). The proteoliposomes were heat treated (37 ºC, 20 min) to deactivate the sample, before being frozen on graphene oxide gold grids. Images (TIFF) were collected with a 300 keV FEI Titan Krios on a Gatan K3 detector in super-resolution mode but bin2 at the hardware stage, using counting mode and a Gatan BioContinuum energy filter (20 eV). The C2 and objective lens apertures were set to 50 and 100 µm, respectively. 3.8 s exposures were captured in 40 frames with a total dose of ~40 Å-2. A pixel size of 1.072 Å/pixel was used.