[{"date_published":"2024-12-01T00:00:00Z","volume":20,"oa_version":"Published Version","publication_status":"published","publisher":"Wiley","tmp":{"short":"CC BY (4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png"},"intvolume":"        20","file_date_updated":"2025-01-29T15:24:50Z","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","publication":"Alzheimer's & Dementia","_id":"18967","date_updated":"2025-01-29T15:29:22Z","article_processing_charge":"Yes (in subscription journal)","issue":"S6","OA_place":"publisher","language":[{"iso":"eng"}],"OA_type":"hybrid","has_accepted_license":"1","publication_identifier":{"issn":["1552-5260"],"eissn":["1552-5279"]},"citation":{"chicago":"Petrova, Olga, Sergey A Trushin, Thi Kim Oanh Nguyen, Mark Ostroot, Matthew Schellenberg, Graham Johnson, Eugenia Trushina, and Leonid A Sazanov. <i>Structure‐activity Relationship Study of Neuroprotective Complex I Inhibitor CP2</i>. <i>Alzheimer’s &#38; Dementia</i>. Vol. 20. Wiley, 2024. <a href=\"https://doi.org/10.1002/alz.085971\">https://doi.org/10.1002/alz.085971</a>.","ieee":"O. Petrova <i>et al.</i>, <i>Structure‐activity relationship study of neuroprotective complex I inhibitor CP2</i>, vol. 20, no. S6. Wiley, 2024.","ama":"Petrova O, Trushin SA, Nguyen TKO, et al. <i>Structure‐activity Relationship Study of Neuroprotective Complex I Inhibitor CP2</i>. Vol 20. Wiley; 2024. doi:<a href=\"https://doi.org/10.1002/alz.085971\">10.1002/alz.085971</a>","mla":"Petrova, Olga, et al. “Structure‐activity Relationship Study of Neuroprotective Complex I Inhibitor CP2.” <i>Alzheimer’s &#38; Dementia</i>, vol. 20, no. S6, e085971, Wiley, 2024, doi:<a href=\"https://doi.org/10.1002/alz.085971\">10.1002/alz.085971</a>.","ista":"Petrova O, Trushin SA, Nguyen TKO, Ostroot M, Schellenberg M, Johnson G, Trushina E, Sazanov LA. 2024. Structure‐activity relationship study of neuroprotective complex I inhibitor CP2, Wiley,p.","short":"O. Petrova, S.A. Trushin, T.K.O. Nguyen, M. Ostroot, M. Schellenberg, G. Johnson, E. Trushina, L.A. Sazanov, Structure‐activity Relationship Study of Neuroprotective Complex I Inhibitor CP2, Wiley, 2024.","apa":"Petrova, O., Trushin, S. A., Nguyen, T. K. O., Ostroot, M., Schellenberg, M., Johnson, G., … Sazanov, L. A. (2024). <i>Structure‐activity relationship study of neuroprotective complex I inhibitor CP2</i>. <i>Alzheimer’s &#38; Dementia</i> (Vol. 20). Wiley. <a href=\"https://doi.org/10.1002/alz.085971\">https://doi.org/10.1002/alz.085971</a>"},"department":[{"_id":"LeSa"}],"status":"public","quality_controlled":"1","type":"other_academic_publication","file":[{"success":1,"date_created":"2025-01-29T15:24:50Z","content_type":"application/pdf","file_name":"2024_AlzheimerDementia_Petrova.pdf","file_id":"18968","file_size":70870,"checksum":"e914bd5f3a701659ab79d497a122811f","date_updated":"2025-01-29T15:24:50Z","creator":"dernst","access_level":"open_access","relation":"main_file"}],"oa":1,"date_created":"2025-01-29T15:21:40Z","abstract":[{"text":"Background: We identified small molecule tricyclic pyrone compound CP2 as a mild mitochondrial complex I (MCI) inhibitor that induces neuroprotection in multiple mouse models of AD. One of the major concerns while targeting mitochondria is the production of reactive oxygen species (ROS). CP2 consists of two diastereoisomers, D1 and D2, with distinct activity and toxicity profiles. This study was designed to understand how structure of D1 and D2 affects their binding to MCI and the consequential impact on ROS production.\r\n\r\nMethod: The X-ray crystallography and cryo-electron microscopy (cryo-EM) at global resolution of 3.25-3.27Å were employed to identify the molecular structure of D1 and D2 and the D1 binding to the isolated ovine MCI. The assessment of the MCI inhibition and the extent of ROS generation were done in isolated MCI and human neuroblastoma MC65 cells using flow cytometry, a Seahorse extracellular flux analyzer, and the kinetic studies.\r\n\r\nResult: In the closed conformation of MCI, D1 selectively binds to the deep Quinone-site (Qd) but not to the shallow Q-site (Qs), sharing the same binding pocket as rotenone. In the open MCI state, D1 exclusively binds to the Qs in contrast to rotenone, which binds Qd and Qs in both closed and open states. At the same concentrations, D1 inhibits respiration to a greater extent compared to D2 (5:1 ratio) and produces higher level of ROS.\r\n\r\nConclusion:Cryo-EM unambiguously identified binding of D1 to both the Qd and Qs sites, contingent upon the conformational state of MCI. In contrast to rotenone, D1 binds Qd only in the closed conformation during catalytic cycle, leading to mild inhibition. Superimposing X-ray crystallography data of D1 and D2 onto cryo-EM data suggests that the orientation of the methyl group in D2 induces a flatter conformation, resulting in lower binding affinity to MCI, which correlates with lower inhibition and toxicity compared to D1. At physiologically relevant concentrations, CP2 (D1:D2 = 1:1) demonstrates low MCI inhibition yielding negligible ROS levels. This observation provides new insight into the absence of toxicity associated with CP2 treatment in vivo, further highlighting feasibility for the development of safe and efficacious MCI inhibitors.","lang":"eng"}],"title":"Structure‐activity relationship study of neuroprotective complex I inhibitor CP2","month":"12","ddc":["570"],"article_number":"e085971","day":"01","author":[{"last_name":"Petrova","first_name":"Olga","full_name":"Petrova, Olga","id":"5D8C9660-5D49-11EA-8188-567B3DDC885E"},{"full_name":"Trushin, Sergey A","last_name":"Trushin","first_name":"Sergey A"},{"first_name":"Thi Kim Oanh","last_name":"Nguyen","full_name":"Nguyen, Thi Kim Oanh"},{"full_name":"Ostroot, Mark","last_name":"Ostroot","first_name":"Mark"},{"full_name":"Schellenberg, Matthew","last_name":"Schellenberg","first_name":"Matthew"},{"full_name":"Johnson, Graham","first_name":"Graham","last_name":"Johnson"},{"first_name":"Eugenia","last_name":"Trushina","full_name":"Trushina, Eugenia"},{"last_name":"Sazanov","first_name":"Leonid A","orcid":"0000-0002-0977-7989","full_name":"Sazanov, Leonid A","id":"338D39FE-F248-11E8-B48F-1D18A9856A87"}],"doi":"10.1002/alz.085971","year":"2024"},{"related_material":{"record":[{"id":"12781","status":"public","relation":"dissertation_contains"}],"link":[{"url":"https://doi.org/10.1038/s41586-022-05457-8","relation":"erratum"},{"relation":"press_release","description":"News on ISTA website","url":"https://ista.ac.at/en/news/proton-dominos-kick-off-life/"}]},"ddc":["572"],"title":"A universal coupling mechanism of respiratory complex I","month":"09","day":"22","page":"808-814","scopus_import":"1","project":[{"grant_number":"25541","_id":"238A0A5A-32DE-11EA-91FC-C7463DDC885E","name":"Structural characterization of E. coli complex I: an important mechanistic model"},{"_id":"627abdeb-2b32-11ec-9570-ec31a97243d3","grant_number":"101020697","call_identifier":"H2020","name":"Structure and mechanism of respiratory chain molecular machines"}],"author":[{"full_name":"Kravchuk, Vladyslav","id":"4D62F2A6-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-9523-9089","first_name":"Vladyslav","last_name":"Kravchuk"},{"id":"5D8C9660-5D49-11EA-8188-567B3DDC885E","full_name":"Petrova, Olga","last_name":"Petrova","first_name":"Olga"},{"orcid":"0000-0002-6018-3422","first_name":"Domen","last_name":"Kampjut","full_name":"Kampjut, Domen","id":"37233050-F248-11E8-B48F-1D18A9856A87"},{"first_name":"Anna","last_name":"Wojciechowska-Bason","full_name":"Wojciechowska-Bason, Anna"},{"last_name":"Breese","first_name":"Zara","full_name":"Breese, Zara"},{"id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A","orcid":"0000-0002-0977-7989","first_name":"Leonid A","last_name":"Sazanov"}],"year":"2022","doi":"10.1038/s41586-022-05199-7","article_type":"original","corr_author":"1","status":"public","quality_controlled":"1","date_created":"2023-01-12T12:04:33Z","abstract":[{"lang":"eng","text":"Complex I is the first enzyme in the respiratory chain, which is responsible for energy production in mitochondria and bacteria1. Complex I couples the transfer of two electrons from NADH to quinone and the translocation of four protons across the membrane2, but the coupling mechanism remains contentious. Here we present cryo-electron microscopy structures of Escherichia coli complex I (EcCI) in different redox states, including catalytic turnover. EcCI exists mostly in the open state, in which the quinone cavity is exposed to the cytosol, allowing access for water molecules, which enable quinone movements. Unlike the mammalian paralogues3, EcCI can convert to the closed state only during turnover, showing that closed and open states are genuine turnover intermediates. The open-to-closed transition results in the tightly engulfed quinone cavity being connected to the central axis of the membrane arm, a source of substrate protons. Consistently, the proportion of the closed state increases with increasing pH. We propose a detailed but straightforward and robust mechanism comprising a ‘domino effect’ series of proton transfers and electrostatic interactions: the forward wave (‘dominoes stacking’) primes the pump, and the reverse wave (‘dominoes falling’) results in the ejection of all pumped protons from the distal subunit NuoL. This mechanism explains why protons exit exclusively from the NuoL subunit and is supported by our mutagenesis data. We contend that this is a universal coupling mechanism of complex I and related enzymes."}],"type":"journal_article","keyword":["Multidisciplinary"],"file":[{"success":1,"content_type":"application/pdf","date_created":"2023-05-30T17:05:31Z","file_name":"EcCxI_manuscript_rev3_noSI_updated_withFigs_opt.pdf","date_updated":"2023-05-30T17:05:31Z","access_level":"open_access","creator":"lsazanov","relation":"main_file","file_id":"13104","file_size":1425655,"checksum":"d42a93e24f59e883ef0b5429832391d0"},{"success":1,"content_type":"application/pdf","date_created":"2023-05-30T17:07:05Z","file_name":"EcCxI_manuscript_rev3_SI_All_opt_upd.pdf","date_updated":"2023-05-30T17:07:05Z","creator":"lsazanov","access_level":"open_access","relation":"main_file","file_id":"13105","file_size":9842513,"checksum":"5422bc0a73b3daadafa262c7ea6deae3"}],"acknowledged_ssus":[{"_id":"EM-Fac"},{"_id":"LifeSc"},{"_id":"ScienComp"}],"oa":1,"issue":"7928","_id":"12138","date_updated":"2026-04-30T22:30:30Z","article_processing_charge":"No","citation":{"apa":"Kravchuk, V., Petrova, O., Kampjut, D., Wojciechowska-Bason, A., Breese, Z., &#38; Sazanov, L. A. (2022). A universal coupling mechanism of respiratory complex I. <i>Nature</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41586-022-05199-7\">https://doi.org/10.1038/s41586-022-05199-7</a>","ista":"Kravchuk V, Petrova O, Kampjut D, Wojciechowska-Bason A, Breese Z, Sazanov LA. 2022. A universal coupling mechanism of respiratory complex I. Nature. 609(7928), 808–814.","short":"V. Kravchuk, O. Petrova, D. Kampjut, A. Wojciechowska-Bason, Z. Breese, L.A. Sazanov, Nature 609 (2022) 808–814.","mla":"Kravchuk, Vladyslav, et al. “A Universal Coupling Mechanism of Respiratory Complex I.” <i>Nature</i>, vol. 609, no. 7928, Springer Nature, 2022, pp. 808–14, doi:<a href=\"https://doi.org/10.1038/s41586-022-05199-7\">10.1038/s41586-022-05199-7</a>.","ama":"Kravchuk V, Petrova O, Kampjut D, Wojciechowska-Bason A, Breese Z, Sazanov LA. A universal coupling mechanism of respiratory complex I. <i>Nature</i>. 2022;609(7928):808-814. doi:<a href=\"https://doi.org/10.1038/s41586-022-05199-7\">10.1038/s41586-022-05199-7</a>","ieee":"V. Kravchuk, O. Petrova, D. Kampjut, A. Wojciechowska-Bason, Z. Breese, and L. A. Sazanov, “A universal coupling mechanism of respiratory complex I,” <i>Nature</i>, vol. 609, no. 7928. Springer Nature, pp. 808–814, 2022.","chicago":"Kravchuk, Vladyslav, Olga Petrova, Domen Kampjut, Anna Wojciechowska-Bason, Zara Breese, and Leonid A Sazanov. “A Universal Coupling Mechanism of Respiratory Complex I.” <i>Nature</i>. Springer Nature, 2022. <a href=\"https://doi.org/10.1038/s41586-022-05199-7\">https://doi.org/10.1038/s41586-022-05199-7</a>."},"publication_identifier":{"eissn":["1476-4687"],"issn":["0028-0836"]},"department":[{"_id":"LeSa"}],"pmid":1,"external_id":{"pmid":["36104567"],"isi":["000854788200001"]},"language":[{"iso":"eng"}],"has_accepted_license":"1","publication_status":"published","publisher":"Springer Nature","date_published":"2022-09-22T00:00:00Z","volume":609,"ec_funded":1,"oa_version":"Submitted Version","intvolume":"       609","file_date_updated":"2023-05-30T17:07:05Z","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","publication":"Nature","acknowledgement":"This research was supported by the Scientific Service Units (SSU) of IST Austria through resources provided by the Electron Microscopy Facility (EMF), the Life Science Facility (LSF) and the IST high-performance computing cluster. We thank V.-V. Hodirnau from IST Austria EMF, M. Babiak from CEITEC for assistance with collecting cryo-EM data and A. Charnagalov for the assistance with protein purification. V.K. was a recipient of a DOC Fellowship of the Austrian Academy of Sciences at the Institute of Science and Technology, Austria. V.K. and O.P. are funded by the ERC Advanced Grant 101020697 RESPICHAIN to L.S. This work was also supported by the Medical Research Council (UK).","isi":1}]