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PreprintCited by 72Mitochondrial Function and PathologyATP Synthase and ATPases ResearchAdvanced Electron Microscopy Techniques and Applications

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Molecular pathway of mitochondrial preprotein import through the TOM-TIM23 supercomplex

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Zhou, Xueyin, Yang, Yuqi, Wang, Guopeng, Wang, Shanshan, Sun, Dongjie, Ou, Xiaomin, Lian, Yuke, Li, Long·2023·Nature Structural & Molecular Biology
DOI 10.1101/2023.06.21.546012

Preprint — published as 10.1038/s41594-023-01103-7 · Paper page

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Abstract

Most mitochondrial proteins need to be imported from the cytosol. Over half of mitochondrial proteins are imported through the pre-sequence pathway that is controlled by the TOM complex in the outer membrane and the TIM23 complex in the inner membrane. It is unclear on the molecular level how proteins cross the mitochondrial double membranes through the TOM and TIM23 complexes. Here, we report the assembly of the active TOM-TIM23 supercomplex with translocating polypeptide substrates captured in the import pathway. Electron cryo-microscopy (Cryo-EM) analyses reveal that during translocation across the outer membrane, the polypeptide substrates pass through the center of the Tom40 channel while interacting with a glutamine-rich patch in the inner wall of Tom40. Structural and biochemical analyses show that the TIM23 complex contains a heterotrimer of the subunits Tim23, Tim17, and Mgr2 in the inner membrane. Tim17 and Mgr2 shield the polypeptide substrates from the lipid environment. The import pathway consists of two highly conserved residue patches of Tim17, one negatively charged patch at the entrance and one hydrophobic patch in the middle of the pathway. These data reveal an unexpected pre-sequence pathway mediated by the TOM-TIM23 supercomplex for facilitating protein import across the double membranes of mitochondria.

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Metadata

Authors
Zhou, X., Yang, Y., Wang, G., Wang, S., Sun, D., Ou, X., Lian, Y., & Li, L.
Affiliations
Peking University (CN)
Corresponding author
Li, Long
ORCID
Venue
Nature Structural & Molecular Biology
Publisher
openRxiv
Published
2023-06-22
Volume
30
Issue
12
Pages
1996-2008
ISSN
1545-9985 (ISSN-L)
DOI
10.1101/2023.06.21.546012
OpenAlex
W4386609897
Cited by
72
References
1
Citations by year
2026: 23 · 2025: 19 · 2024: 28 · 2023: 2
FWCI
6.68
Type
journal article
Language
en
Open access
Licence
cc-by-nc-nd
Locations
  • Nature Structural & Molecular Biology (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • National Natural Science Foundation of China (31870835)
  • Peking University
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
Keywords
TOM complex, TIM23 complex, Tom40, Tim23, Tim17, mitochondrial protein translocation, Saccharomyces cerevisiae
MeSH terms
Mitochondrial Precursor Protein Import Complex Proteins, Carrier Proteins, Membrane Proteins, Mitochondria, Peptides, Saccharomyces cerevisiae, Protein Transport, Mitochondrial Proteins, Membrane Transport Proteins, Saccharomyces cerevisiae Proteins, Mitochondrial Membrane Transport Proteins

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