{"EMPIAR-11663":{"imagesets":[{"segmentations":[],"name":"Unaligned multi-frame micrographs of NUCM R121M Yarrowia lipolytica mitochondrial complex I from an as-prepared sample solubilised in DDM.","directory":"data","category":"micrographs - multiframe","header_format":"MRC","data_format":"MRC","num_images_or_tilt_series":2241,"frames_per_image":40,"frame_range_min":1,"frame_range_max":40,"voxel_type":"32 BIT FLOAT","pixel_width":1.07,"pixel_height":1.07,"micrographs_file_pattern":"","picked_particles_file_pattern":"","picked_particles_directory":"","details":"Unaligned multi-frame micrographs of NUCM R121M Yarrowia lipolytica mitochondrial complex I from an as-prepared sample solubilised in DDM. Frozen gold grids treated with PEGylated thiol reagent. Images (MRC) were collected with a 300 keV FEI Titan Krios on a Gatan K2 Summit detector, using counting mode and a Gatan BioContinuum energy filter (20 eV). 10 s exposures were captured in 40 frames with a total dose of ~48 e/Å-2. A nominal pixel size of 1.07 Å/pixel was used (calibrated 1.05 Å/pixel using pre-existing models).","image_width":"3838","image_height":"3710"}],"workflow_file":null,"grant_references":[{"funding_body":"Medical Research Council (MRC, United Kingdom)","code":"MC_U105663141","country":"United Kingdom"},{"funding_body":"Medical Research Council (MRC, United Kingdom)","code":"MC_UU_00015/2","country":"United Kingdom"},{"funding_body":"Engineering and Physical Sciences Research Council","code":"EP/M024393/1","country":"United Kingdom"},{"funding_body":"Engineering and Physical Sciences Research Council","code":"EP/ T031425/1","country":"United Kingdom"}],"version_history":[],"title":"Single particle cryo-EM dataset of Yarrowia lipolytica mitochondrial complex I with R121M NDUFS2/NUCM mutation","principal_investigator":[{"author_orcid":"0000-0001-8667-6797","middle_name":null,"organization":"MRC Mitochondrial Biology Unit","street":"Hills Road","town_or_city":"Cambridge","state_or_province":"Cambridgeshire","post_or_zip":"CB2 0XY","telephone":null,"fax":null,"first_name":"Judy","last_name":"Hirst","email":"jh [at] mrc-mbu.cam.ac.uk","country":"United Kingdom","entry":"EMPIAR-11663"}],"status":"REL","deposition_date":"2023-08-02","release_date":"2024-06-27","obsolete_date":null,"update_date":"2024-06-27","corresponding_author":{"author":{"author_orcid":"0000-0001-8667-6797","middle_name":null,"organization":"MRC Mitochondrial Biology Unit","street":"Hills Road","town_or_city":"Cambridge","state_or_province":"Cambridgeshire","post_or_zip":"CB2 0XY","first_name":"Judy","last_name":"Hirst","country":"United Kingdom"}},"authors":[{"author":{"name":"Hameedi MA","author_orcid":null}},{"author":{"name":"Grba DN","author_orcid":"0000-0003-2915-951X"}},{"author":{"name":"Richardson KH","author_orcid":null}},{"author":{"name":"Jones AJY","author_orcid":null}},{"author":{"name":"Song W","author_orcid":null}},{"author":{"name":"Roessler MM","author_orcid":"0000-0002-5291-4328"}},{"author":{"name":"Wright JJ","author_orcid":"0000-0001-8987-4449"}},{"author":{"name":"Hirst J","author_orcid":"0000-0001-8667-6797"}}],"cross_references":["EMD-11969"],"biostudies_references":[],"idr_references":[],"empiar_references":[],"citation":[{"authors":[{"name":"Hameedi MA","author_orcid":null},{"name":"Grba DN","author_orcid":"0000-0003-2915-951X"},{"name":"Richardson KH","author_orcid":"0000-0002-5166-0100"},{"name":"Jones AJY","author_orcid":null},{"name":"Song W","author_orcid":null},{"name":"Roessler MM","author_orcid":"0000-0002-5291-4328"},{"name":"Wright JJ","author_orcid":"0000-0001-8987-4449"},{"name":"Hirst J","author_orcid":"0000-0001-8667-6797"}],"editors":[],"published":true,"j_or_nj_citation":true,"title":"A conserved arginine residue is critical for stabilizing the N2 FeS cluster in mitochondrial complex I","volume":"296","country":"","first_page":null,"last_page":null,"year":"2021","language":"English","doi":"10.1016/j.jbc.2021.100474","pubmedid":"33640456","details":"Respiratory complex I (NADH:ubiquinone oxidoreductase), the first enzyme of the electron-transport chain, captures the free energy released by NADH oxidation and ubiquinone reduction to translocate protons across an energy-transducing membrane and drive ATP synthesis during oxidative phosphorylation. The cofactor that transfers the electrons directly to ubiquinone is an iron-sulfur cluster (N2) located in the NDUFS2/NUCM subunit. A nearby arginine residue (R121), which forms part of the second coordination sphere of the N2 cluster, is known to be post-translationally dimethylated but its functional and structural significance are not known. Here, we show that mutations of this arginine residue (R121M/K) abolish the quinone-reductase activity, concomitant with disappearance of the N2 signature from the electron paramagnetic resonance (EPR) spectrum. Analysis of the cryo-EM structure of NDUFS2-R121M complex I at 3.7 Å resolution identified the absence of the cubane N2 cluster as the cause of the dysfunction, within an otherwise intact enzyme. The mutation further induced localized disorder in nearby elements of the quinone-binding site, consistent with the close connections between the cluster and substrate-binding regions. Our results demonstrate that R121 is required for the formation and/or stability of the N2 cluster and highlight the importance of structural analyses for mechanistic interpretation of biochemical and spectroscopic data on complex I variants.","book_chapter_title":null,"publisher":null,"publication_location":null,"journal":"The Journal of biological chemistry","journal_abbreviation":"J Biol Chem","issue":null,"preprint":false}],"dataset_size":"4.6 TB","experiment_type":"EMDB","scale":"molecule","related_pdb_entries":["7b0n"],"entry_doi":"10.6019/EMPIAR-11663"}}