Fluorescence Mapping of Mitochondrial TIM23 Complex Reveals a Water-Facing, Substrate-Interacting Helix Surface

Nathan N. Alder, Robert E. Jensen, Arthur E. Johnson

Research output: Contribution to journalArticle

Abstract

Protein translocation across the mitochondrial inner membrane is mediated by the TIM23 complex. While its central component, Tim23, is believed to form a protein-conducting channel, the regions of this subunit that face the imported protein are unknown. To examine Tim23 structure and environment in intact membranes at high resolution, various derivatives, each with a single, environment-sensitive fluorescent probe positioned at a specific site, were assembled into functional TIM23 complexes in active mitochondria and analyzed by multiple spectral techniques. Probes placed sequentially throughout a transmembrane region that was identified by crosslinking as part of the protein-conducting channel revealed an α helix in an amphipathic environment. Probes on the aqueous-facing helical surface specifically underwent spectral changes during protein import, and their accessibility to hydrophilic quenching agents is considered in terms of channel gating. This approach has therefore provided an unprecedented view of a translocon channel structure in an intact, fully operational, membrane-embedded complex.

Original languageEnglish (US)
Pages (from-to)439-450
Number of pages12
JournalCell
Volume134
Issue number3
DOIs
StatePublished - Aug 8 2008

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Facings
Fluorescence
Water
Substrates
Proteins
Membranes
Mitochondrial Membranes
Protein Transport
Fluorescent Dyes
Mitochondria
Crosslinking
Quenching
Derivatives

Keywords

  • CELLBIO
  • PROTEINS

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Fluorescence Mapping of Mitochondrial TIM23 Complex Reveals a Water-Facing, Substrate-Interacting Helix Surface. / Alder, Nathan N.; Jensen, Robert E.; Johnson, Arthur E.

In: Cell, Vol. 134, No. 3, 08.08.2008, p. 439-450.

Research output: Contribution to journalArticle

Alder, Nathan N. ; Jensen, Robert E. ; Johnson, Arthur E. / Fluorescence Mapping of Mitochondrial TIM23 Complex Reveals a Water-Facing, Substrate-Interacting Helix Surface. In: Cell. 2008 ; Vol. 134, No. 3. pp. 439-450.
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