Autocatalytic processing of the ATP-dependent PIM1 protease: crucial function of a pro-region for sorting to mitochondria

EMBO J. 1997 Dec 15;16(24):7317-25. doi: 10.1093/emboj/16.24.7317.


The biogenesis of the ATP-dependent PIM1 protease of mitochondria was studied by mutational analysis. The ATPase and proteolytic activities of PIM1 were shown to be essential for mitochondrial function. A proteolytically inactive mutant form of PIM1 protease accumulated as a pro-form in mitochondria, revealing a two-step processing of PIM1: the matrix targeting signal is removed by the mitochondrial processing peptidase and then a pro-region of 61 amino acids is cleaved off in an autocatalytic reaction. This latter process depended on the ATP-dependent assembly of PIM1 protease subunits and can occur by an intermolecular and, most probably, also an intramolecular pathway. The respiratory competence of cells harboring mutant PIM1 protease lacking the pro-region was strongly impaired. Subcellular fractionation revealed a cytosolic localization of mutant PIM1 protease. This demonstrates the requirement for the propeptide for efficient sorting of PIM1 protease to mitochondria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Dependent Proteases
  • Adenosine Triphosphate / metabolism*
  • Amino Acid Sequence
  • Catalysis
  • Chromatography, Affinity
  • Chromatography, Gel
  • Cloning, Molecular
  • DNA Primers
  • Enzyme Precursors / chemistry
  • Enzyme Precursors / metabolism*
  • Mitochondria / enzymology*
  • Mitochondrial Proteins
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Polymerase Chain Reaction
  • Protein Processing, Post-Translational*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae Proteins*
  • Serine Endopeptidases / chemistry
  • Serine Endopeptidases / isolation & purification
  • Serine Endopeptidases / metabolism*


  • DNA Primers
  • Enzyme Precursors
  • Mitochondrial Proteins
  • Recombinant Proteins
  • Saccharomyces cerevisiae Proteins
  • Adenosine Triphosphate
  • ATP-Dependent Proteases
  • PIM1 protein, S cerevisiae
  • Serine Endopeptidases