---
OA_place: publisher
OA_type: hybrid
_id: '18967'
abstract:
- lang: eng
  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."
article_number: e085971
article_processing_charge: Yes (in subscription journal)
author:
- first_name: Olga
  full_name: Petrova, Olga
  id: 5D8C9660-5D49-11EA-8188-567B3DDC885E
  last_name: Petrova
- first_name: Sergey A
  full_name: Trushin, Sergey A
  last_name: Trushin
- first_name: Thi Kim Oanh
  full_name: Nguyen, Thi Kim Oanh
  last_name: Nguyen
- first_name: Mark
  full_name: Ostroot, Mark
  last_name: Ostroot
- first_name: Matthew
  full_name: Schellenberg, Matthew
  last_name: Schellenberg
- first_name: Graham
  full_name: Johnson, Graham
  last_name: Johnson
- first_name: Eugenia
  full_name: Trushina, Eugenia
  last_name: Trushina
- first_name: Leonid A
  full_name: Sazanov, Leonid A
  id: 338D39FE-F248-11E8-B48F-1D18A9856A87
  last_name: Sazanov
  orcid: 0000-0002-0977-7989
citation:
  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>
  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>
  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.
  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.
  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>.
  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.
date_created: 2025-01-29T15:21:40Z
date_published: 2024-12-01T00:00:00Z
date_updated: 2025-01-29T15:29:22Z
day: '01'
ddc:
- '570'
department:
- _id: LeSa
doi: 10.1002/alz.085971
file:
- access_level: open_access
  checksum: e914bd5f3a701659ab79d497a122811f
  content_type: application/pdf
  creator: dernst
  date_created: 2025-01-29T15:24:50Z
  date_updated: 2025-01-29T15:24:50Z
  file_id: '18968'
  file_name: 2024_AlzheimerDementia_Petrova.pdf
  file_size: 70870
  relation: main_file
  success: 1
file_date_updated: 2025-01-29T15:24:50Z
has_accepted_license: '1'
intvolume: '        20'
issue: S6
language:
- iso: eng
month: '12'
oa: 1
oa_version: Published Version
publication: Alzheimer's & Dementia
publication_identifier:
  eissn:
  - 1552-5279
  issn:
  - 1552-5260
publication_status: published
publisher: Wiley
quality_controlled: '1'
status: public
title: Structure‐activity relationship study of neuroprotective complex I inhibitor
  CP2
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: other_academic_publication
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 20
year: '2024'
...
---
_id: '12138'
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.'
acknowledged_ssus:
- _id: EM-Fac
- _id: LifeSc
- _id: ScienComp
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).
article_processing_charge: No
article_type: original
author:
- first_name: Vladyslav
  full_name: Kravchuk, Vladyslav
  id: 4D62F2A6-F248-11E8-B48F-1D18A9856A87
  last_name: Kravchuk
  orcid: 0000-0001-9523-9089
- first_name: Olga
  full_name: Petrova, Olga
  id: 5D8C9660-5D49-11EA-8188-567B3DDC885E
  last_name: Petrova
- first_name: Domen
  full_name: Kampjut, Domen
  id: 37233050-F248-11E8-B48F-1D18A9856A87
  last_name: Kampjut
  orcid: 0000-0002-6018-3422
- first_name: Anna
  full_name: Wojciechowska-Bason, Anna
  last_name: Wojciechowska-Bason
- first_name: Zara
  full_name: Breese, Zara
  last_name: Breese
- first_name: Leonid A
  full_name: Sazanov, Leonid A
  id: 338D39FE-F248-11E8-B48F-1D18A9856A87
  last_name: Sazanov
  orcid: 0000-0002-0977-7989
citation:
  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>
  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>
  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>.
  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.
  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.
  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>.
  short: V. Kravchuk, O. Petrova, D. Kampjut, A. Wojciechowska-Bason, Z. Breese, L.A.
    Sazanov, Nature 609 (2022) 808–814.
corr_author: '1'
date_created: 2023-01-12T12:04:33Z
date_published: 2022-09-22T00:00:00Z
date_updated: 2026-06-21T22:30:38Z
day: '22'
ddc:
- '572'
department:
- _id: LeSa
doi: 10.1038/s41586-022-05199-7
ec_funded: 1
external_id:
  isi:
  - '000854788200001'
  pmid:
  - '36104567'
file:
- access_level: open_access
  checksum: d42a93e24f59e883ef0b5429832391d0
  content_type: application/pdf
  creator: lsazanov
  date_created: 2023-05-30T17:05:31Z
  date_updated: 2023-05-30T17:05:31Z
  file_id: '13104'
  file_name: EcCxI_manuscript_rev3_noSI_updated_withFigs_opt.pdf
  file_size: 1425655
  relation: main_file
  success: 1
- access_level: open_access
  checksum: 5422bc0a73b3daadafa262c7ea6deae3
  content_type: application/pdf
  creator: lsazanov
  date_created: 2023-05-30T17:07:05Z
  date_updated: 2023-05-30T17:07:05Z
  file_id: '13105'
  file_name: EcCxI_manuscript_rev3_SI_All_opt_upd.pdf
  file_size: 9842513
  relation: main_file
  success: 1
file_date_updated: 2023-05-30T17:07:05Z
has_accepted_license: '1'
intvolume: '       609'
isi: 1
issue: '7928'
keyword:
- Multidisciplinary
language:
- iso: eng
month: '09'
oa: 1
oa_version: Submitted Version
page: 808-814
pmid: 1
project:
- _id: 238A0A5A-32DE-11EA-91FC-C7463DDC885E
  grant_number: '25541'
  name: 'Structural characterization of E. coli complex I: an important mechanistic
    model'
- _id: 627abdeb-2b32-11ec-9570-ec31a97243d3
  call_identifier: H2020
  grant_number: '101020697'
  name: Structure and mechanism of respiratory chain molecular machines
publication: Nature
publication_identifier:
  eissn:
  - 1476-4687
  issn:
  - 0028-0836
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
related_material:
  link:
  - relation: erratum
    url: https://doi.org/10.1038/s41586-022-05457-8
  - description: News on ISTA website
    relation: press_release
    url: https://ista.ac.at/en/news/proton-dominos-kick-off-life/
  record:
  - id: '12781'
    relation: dissertation_contains
    status: public
scopus_import: '1'
status: public
title: A universal coupling mechanism of respiratory complex I
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 609
year: '2022'
...