Mapping protein interactions in the active TOM-TIM23 supercomplex.
Cell Fractionation
Cell Nucleus
/ metabolism
Cross-Linking Reagents
/ chemistry
Mass Spectrometry
/ methods
Membrane Transport Proteins
/ chemistry
Mitochondria
/ metabolism
Mitochondrial Membrane Transport Proteins
/ chemistry
Mitochondrial Membranes
/ metabolism
Mitochondrial Precursor Protein Import Complex Proteins
Molecular Docking Simulation
Mutagenesis, Site-Directed
Point Mutation
Protein Binding
/ genetics
Protein Interaction Mapping
/ methods
Protein Precursors
/ chemistry
Recombinant Proteins
/ chemistry
Saccharomyces cerevisiae Proteins
/ chemistry
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
29 09 2021
29 09 2021
Historique:
received:
22
12
2020
accepted:
19
08
2021
entrez:
30
9
2021
pubmed:
1
10
2021
medline:
24
10
2021
Statut:
epublish
Résumé
Nuclear-encoded mitochondrial proteins destined for the matrix have to be transported across two membranes. The TOM and TIM23 complexes facilitate the transport of precursor proteins with N-terminal targeting signals into the matrix. During transport, precursors are recognized by the TIM23 complex in the inner membrane for handover from the TOM complex. However, we have little knowledge on the organization of the TOM-TIM23 transition zone and on how precursor transfer between the translocases occurs. Here, we have designed a precursor protein that is stalled during matrix transport in a TOM-TIM23-spanning manner and enables purification of the translocation intermediate. Combining chemical cross-linking with mass spectrometric analyses and structural modeling allows us to map the molecular environment of the intermembrane space interface of TOM and TIM23 as well as the import motor interactions with amino acid resolution. Our analyses provide a framework for understanding presequence handover and translocation during matrix protein transport.
Identifiants
pubmed: 34588454
doi: 10.1038/s41467-021-26016-1
pii: 10.1038/s41467-021-26016-1
pmc: PMC8481542
doi:
Substances chimiques
Cross-Linking Reagents
0
Membrane Transport Proteins
0
Mitochondrial Membrane Transport Proteins
0
Mitochondrial Precursor Protein Import Complex Proteins
0
Protein Precursors
0
Recombinant Proteins
0
Saccharomyces cerevisiae Proteins
0
TIM23 protein, S cerevisiae
0
TIM44 protein, S cerevisiae
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
5715Informations de copyright
© 2021. The Author(s).
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