---
_id: '1209'
abstract:
- lang: eng
  text: 'NADH-ubiquinone oxidoreductase (complex I) is the largest (∼1 MDa) and the
    least characterized complex of the mitochondrial electron transport chain. Because
    of the ease of sample availability, previous work has focused almost exclusively
    on bovine complex I. However, only medium resolution structural analyses of this
    complex have been reported. Working with other mammalian complex I homologues
    is a potential approach for overcoming these limitations. Due to the inherent
    difficulty of expressing large membrane protein complexes, screening of complex
    I homologues is limited to large mammals reared for human consumption. The high
    sequence identity among these available sources may preclude the benefits of screening.
    Here, we report the characterization of complex I purified from Ovis aries (ovine)
    heart mitochondria. All 44 unique subunits of the intact complex were identified
    by mass spectrometry. We identified differences in the subunit composition of
    subcomplexes of ovine complex I as compared with bovine, suggesting differential
    stability of inter-subunit interactions within the complex. Furthermore, the 42-kDa
    subunit, which is easily lost from the bovine enzyme, remains tightly bound to
    ovine complex I. Additionally, we developed a novel purification protocol for
    highly active and stable mitochondrial complex I using the branched-chain detergent
    lauryl maltose neopentyl glycol. Our data demonstrate that, although closely related,
    significant differences exist between the biochemical properties of complex I
    prepared from ovine and bovine mitochondria and that ovine complex I represents
    a suitable alternative target for further structural studies. '
acknowledgement: "J.A.S supported in part by a Medical Research D.G.Council UK Ph.D.
  fellowship.\r\nThis work was supported in part by European Union's 2020 Research
  and Innovation Program under Grant 701309. \r\n"
article_processing_charge: No
author:
- first_name: James A
  full_name: Letts, James A
  id: 322DA418-F248-11E8-B48F-1D18A9856A87
  last_name: Letts
  orcid: 0000-0002-9864-3586
- first_name: Gianluca
  full_name: Degliesposti, Gianluca
  last_name: Degliesposti
- first_name: Karol
  full_name: Fiedorczuk, Karol
  id: 5BFF67CE-02D1-11E9-B11A-A5A4D7DFFFD0
  last_name: Fiedorczuk
- first_name: Mark
  full_name: Skehel, Mark
  last_name: Skehel
- 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: Letts JA, Degliesposti G, Fiedorczuk K, Skehel M, Sazanov LA. Purification
    of ovine respiratory complex i results in a highly active and stable preparation.
    <i>Journal of Biological Chemistry</i>. 2016;291(47):24657-24675. doi:<a href="https://doi.org/10.1074/jbc.M116.735142">10.1074/jbc.M116.735142</a>
  apa: Letts, J. A., Degliesposti, G., Fiedorczuk, K., Skehel, M., &#38; Sazanov,
    L. A. (2016). Purification of ovine respiratory complex i results in a highly
    active and stable preparation. <i>Journal of Biological Chemistry</i>. American
    Society for Biochemistry and Molecular Biology. <a href="https://doi.org/10.1074/jbc.M116.735142">https://doi.org/10.1074/jbc.M116.735142</a>
  chicago: Letts, James A, Gianluca Degliesposti, Karol Fiedorczuk, Mark Skehel, and
    Leonid A Sazanov. “Purification of Ovine Respiratory Complex i Results in a Highly
    Active and Stable Preparation.” <i>Journal of Biological Chemistry</i>. American
    Society for Biochemistry and Molecular Biology, 2016. <a href="https://doi.org/10.1074/jbc.M116.735142">https://doi.org/10.1074/jbc.M116.735142</a>.
  ieee: J. A. Letts, G. Degliesposti, K. Fiedorczuk, M. Skehel, and L. A. Sazanov,
    “Purification of ovine respiratory complex i results in a highly active and stable
    preparation,” <i>Journal of Biological Chemistry</i>, vol. 291, no. 47. American
    Society for Biochemistry and Molecular Biology, pp. 24657–24675, 2016.
  ista: Letts JA, Degliesposti G, Fiedorczuk K, Skehel M, Sazanov LA. 2016. Purification
    of ovine respiratory complex i results in a highly active and stable preparation.
    Journal of Biological Chemistry. 291(47), 24657–24675.
  mla: Letts, James A., et al. “Purification of Ovine Respiratory Complex i Results
    in a Highly Active and Stable Preparation.” <i>Journal of Biological Chemistry</i>,
    vol. 291, no. 47, American Society for Biochemistry and Molecular Biology, 2016,
    pp. 24657–75, doi:<a href="https://doi.org/10.1074/jbc.M116.735142">10.1074/jbc.M116.735142</a>.
  short: J.A. Letts, G. Degliesposti, K. Fiedorczuk, M. Skehel, L.A. Sazanov, Journal
    of Biological Chemistry 291 (2016) 24657–24675.
corr_author: '1'
date_created: 2018-12-11T11:50:44Z
date_published: 2016-11-18T00:00:00Z
date_updated: 2025-09-22T09:38:04Z
day: '18'
department:
- _id: LeSa
doi: 10.1074/jbc.M116.735142
ec_funded: 1
external_id:
  isi:
  - '000388875500026'
intvolume: '       291'
isi: 1
issue: '47'
language:
- iso: eng
main_file_link:
- open_access: '1'
  url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5114416/
month: '11'
oa: 1
oa_version: Submitted Version
page: 24657 - 24675
project:
- _id: 2593EBD6-B435-11E9-9278-68D0E5697425
  name: Atomic-Resolution Structures of Mitochondrial Respiratory Chain Supercomplexes
- _id: 2590DB08-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '701309'
  name: Atomic Resolution Structures of Mitochondrial Respiratory Chain Supercomplexes
publication: Journal of Biological Chemistry
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '6139'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Purification of ovine respiratory complex i results in a highly active and
  stable preparation
type: journal_article
user_id: 317138e5-6ab7-11ef-aa6d-ffef3953e345
volume: 291
year: '2016'
...