Analysis of elements in the substrate required for processing by mitochondrial processing peptidase

Tadashi Ogishima, Takuro Niidome, Kunitoshi Shimokata, Sakae Kitada, Akio Ito

Research output: Contribution to journalArticle

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Abstract

We have recently demonstrated that synthetic peptides modeled on the extension peptide of malate dehydrogenase can be a good substrate of mitochondrial processing peptidase and that arginine residues present at positions -2 or -3 and distant from the cleavage point were important for recognition by the enzyme (Niidome, T., Kitada, S., Shimokata, K., Ogishima, T., and Ito, A. (1994) J. Biol. Chem. 269, 24719-24722). We further investigated the elements required for substrates of the protease. To analyze the reaction by a more rapid yet quantitative method, we have developed intramolecularly quenched fluorescent substrates. Using the fluorogenic substrates we demonstrated that at least one of the proline and glycine between the distal and proximal arginine residues was also important while other connecting sequences were dispensable. In addition, the protease showed considerable preference for aromatic and, to a lesser extent, hydrophobic amino acids in the P1'-position. These results together with the previous daryl suggest that the proximal and distal arginine residues, proline and/or glycine between them, and P1' amino acid could be critical determinants for the specific cleavage of the substrates by the protease.

Original languageEnglish
Pages (from-to)30322-30326
Number of pages5
JournalJournal of Biological Chemistry
Volume270
Issue number51
DOIs
Publication statusPublished - Dec 22 1995

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Arginine
Peptide Hydrolases
Proline
Glycine
Substrates
Processing
Amino Acids
Malate Dehydrogenase
Peptides
Fluorescent Dyes
Enzymes
mitochondrial processing peptidase

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Analysis of elements in the substrate required for processing by mitochondrial processing peptidase. / Ogishima, Tadashi; Niidome, Takuro; Shimokata, Kunitoshi; Kitada, Sakae; Ito, Akio.

In: Journal of Biological Chemistry, Vol. 270, No. 51, 22.12.1995, p. 30322-30326.

Research output: Contribution to journalArticle

Ogishima, Tadashi ; Niidome, Takuro ; Shimokata, Kunitoshi ; Kitada, Sakae ; Ito, Akio. / Analysis of elements in the substrate required for processing by mitochondrial processing peptidase. In: Journal of Biological Chemistry. 1995 ; Vol. 270, No. 51. pp. 30322-30326.
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