title: Electrostatic recognition of matrix targeting signal by mitochondrial processing peptidase.
abstract: Mitochondrial processing peptidase (MPP), a metalloendopeptidase consisting of alpha and beta subunits, specifically recognizes a large variety of mitochondrial precursor proteins and cleaves off the N-terminal basic matrix targeting signals (MTS). Basic residues in MTS and acidic sites in MPP are required for effective processing. To elucidate whether the enzyme recognizes the MTS through electrostatic interaction, we investigated the effects of various salts on MPP activity. Decreases in the activity depended on the ionic strength and increases in the Michaelis constant value correlated clearly with the ionic strength, indicating a lower affinity of the enzyme for the substrate. Direct determination of the affinity between MPP and a MTS peptide using surface plasmon resonance showed a decrease in the association rate with high salt and that dissociation constant values were decreased. The effect of salt on the processing activity towards a variety of precursors was confirmed using five precursors with different sequences and lengths of MTS. Thus, we propose that electrostatic interactions are indispensable for the association between various MTS and MPP.
authors.affiliation: Department of Chemistry, Faculty of Science, Kyushu University, Fukuoka 812-8581, Japan. s.kitscc@mbox.nc.kyushu-u.ac.jp
authors.forename: S
authors.initials: S
authors.lastname: Kitada
authors.forename: A
authors.initials: A
authors.lastname: Ito
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D007477:Ions / Q000378:metabolism; D007700:Kinetics; D008666:Metalloendopeptidases / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D008930:Mitochondria, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction / Q000502:physiology*; D013379:Substrate Specificity; D020349:Surface Plasmon Resonance; D000096122:Mitochondrial Processing Peptidase
chemical_list: D007477:Ions; D008666:Metalloendopeptidases; D000096122:Mitochondrial Processing Peptidase