Mutational analysis of both subunits from rat mitochondrial processing peptidase

Hans Martin Striebel, Petr Rysavy, Jiri Adamec, Jaroslav Spizek, Frantisek Kalousek

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Rat liver mitochondrial processing peptidase (MPP) is the primary peptidase that cleaves leader peptides from nuclearly encoded mitochondrial proteins following their transport from the cytosol to the mitochondrial matrix. This enzyme consists of two nonidentical subunits that have overall similarity to each other and share certain amino acid motifs. These include the putative metal-ion binding HFLEH motif in the β-subunit and the HFLEK motif of the α-subunit, as well as a possibly helical amino acid stretch bearing a high concentration of negatively charged residues about 70 amino acids downstream of these motifs in both subunits. In order to achieve a better understanding of the role of certain amino acids in rat MPP, we performed site-directed mutagenesis on both of its subunits. Our results show that whereas both histidines and the glutamate of the HFLEH motif in the β- subunit are crucial for MPP function, this holds true only for the glutamate in the related HFLEK motif in the α-subunit. In addition, functionally important negatively charged residues in the region 70 amino acids downstream occur only in the β-subunit and not in the α-subunit. This indicates a functional asymmetry between the subunits, with the β-subunit containing a majority of residues participating in the active center.

Original languageEnglish (US)
Pages (from-to)211-218
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume335
Issue number1
DOIs
StatePublished - Nov 1 1996

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Rats
Amino Acid Motifs
Amino Acids
Glutamic Acid
Mitochondrial Proteins
Protein Sorting Signals
Site-Directed Mutagenesis
Histidine
Bearings (structural)
Cytosol
Peptide Hydrolases
Metals
Mutagenesis
Ions
Liver
Metal ions
Enzymes
mitochondrial processing peptidase

Keywords

  • active site
  • mitochondrial processing peptidase
  • mutagenesis

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Mutational analysis of both subunits from rat mitochondrial processing peptidase. / Striebel, Hans Martin; Rysavy, Petr; Adamec, Jiri; Spizek, Jaroslav; Kalousek, Frantisek.

In: Archives of Biochemistry and Biophysics, Vol. 335, No. 1, 01.11.1996, p. 211-218.

Research output: Contribution to journalArticle

Striebel, Hans Martin ; Rysavy, Petr ; Adamec, Jiri ; Spizek, Jaroslav ; Kalousek, Frantisek. / Mutational analysis of both subunits from rat mitochondrial processing peptidase. In: Archives of Biochemistry and Biophysics. 1996 ; Vol. 335, No. 1. pp. 211-218.
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