Biogenesis of porin of the outer mitochondrial membrane involves an import pathway via receptors and the general import pore of the TOM complex

Thomas Krimmer, Doron Rapaport, Michael T. Ryan, Chris Meisinger, C. Kenneth Kassenbrock, Elizabeth Blachly-Dyson, Michael Forte, Michael G. Douglas, Walter Neupert, Frank E. Nargang, Nikolaus Pfanner

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131 Scopus citations

Abstract

Porin, also termed the voltage-dependent anion channel, is the most abundant protein of the mitochondrial outer membrane. The process of import, and assembly of the protein is known to be dependent on the surface receptor Tom20, but the requirement for other mitochondrial proteins remains controversial. We have used mitochondria from Neurospora crassa and Saccharomyces cerevisiae to analyze the import pathway of porin. Import of porin into isolated mitochondria in which the outer membrane has been opened is inhibited despite similar levels of Tom20 as in intact mitochondria. A matrix-destined precursor and the porin precursor compete for the same translocation sites in both normal mitochondria and mitochondria whose surface receptors have been removed, suggesting that both precursors utilize the general import pore. Using an assay established to monitor the assembly of in vitro-imported porin into preexisting porin complexes we have shown that besides Tom20, the biogenesis of porin depends on the central receptor Tom22, as well as Tom5 and Tom7 of the general import pore complex (translocase of the outer mitochondrial membrane [TOM] core complex). The characterization of two new mutant alleles of the essential pore protein Tom40 demonstrates that the import of porin also requires a functional Tom40. Moreover, the porin precursor can be cross-linked to Tom20, Tom22, and Tom40 on its import pathway. We conclude that import of porin does not proceed through the action of Tom20 alone, but requires an intact outer membrane and involves at least four more subunits of the TOM machinery, including the general import pore.

Original languageEnglish (US)
Pages (from-to)289-300
Number of pages12
JournalJournal of Cell Biology
Volume152
Issue number2
DOIs
StatePublished - Jan 22 2001

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Keywords

  • Mitochondria
  • Neurospbra crassa
  • Porin
  • Protein sorting
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • Cell Biology

Cite this

Krimmer, T., Rapaport, D., Ryan, M. T., Meisinger, C., Kassenbrock, C. K., Blachly-Dyson, E., Forte, M., Douglas, M. G., Neupert, W., Nargang, F. E., & Pfanner, N. (2001). Biogenesis of porin of the outer mitochondrial membrane involves an import pathway via receptors and the general import pore of the TOM complex. Journal of Cell Biology, 152(2), 289-300. https://doi.org/10.1083/jcb.152.2.289