Two-step processing of human frataxin by mitochondrial processing peptidase: Precursor and intermediate forms are cleaved at different rates

Patrizia Cavadini, Jiri Adamec, Franco Taroni, Oleksandr Gakh, Grazia Isaya

Research output: Contribution to journalArticle

88 Citations (Scopus)

Abstract

We showed previously that maturation of the human frataxin precursor (p-fxn) involves two cleavages by the mitochondrial processing peptidase (MPP). This observation was not confirmed by another group, however, who reported only one cleavage. Here, we demonstrate conclusively that MPP cleaves p-fxn in two sequential steps, yielding a 18,826-Da intermediate (i-fxn) and a 17,255-Da mature (m-fxn) form, the latter corresponding to endogenous frataxin in human tissues. The two cleavages occur between residues 41-42 and 55-56, and both match the MPP consensus sequence RX ↓ (X/S). Recombinant rat and yeast MPP catalyze the p → i step 4 and 40 times faster, respectively, than the i → m step. In isolated rat mitochondria, p-fxn undergoes a sequence of cleavages, p → i → m → d1 → d2, with d1 and d2 representing two C-terminal fragments of m-fxn produced by an unknown protease. The i → m step is limiting, and the overall rate of p → i → m does not exceed the rate of m → d1 → d2, such that the levels of m-fxn do not change during incubations as long as 3 h. Inhibition of the i → m step by a disease-causing frataxin mutation (W173G) leads to nonspecific degradation of i-fxn. Thus, the second of the two processing steps catalyzed by MPP limits the levels of mature frataxin within mitochondria.

Original languageEnglish (US)
Pages (from-to)41469-41475
Number of pages7
JournalJournal of Biological Chemistry
Volume275
Issue number52
DOIs
StatePublished - Dec 29 2000

Fingerprint

Processing
Mitochondria
Rats
Consensus Sequence
Yeast
Peptide Hydrolases
Yeasts
mitochondrial processing peptidase
frataxin
Tissue
Degradation
Mutation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Two-step processing of human frataxin by mitochondrial processing peptidase : Precursor and intermediate forms are cleaved at different rates. / Cavadini, Patrizia; Adamec, Jiri; Taroni, Franco; Gakh, Oleksandr; Isaya, Grazia.

In: Journal of Biological Chemistry, Vol. 275, No. 52, 29.12.2000, p. 41469-41475.

Research output: Contribution to journalArticle

@article{b531fec00ba24664ad7e550034b3b364,
title = "Two-step processing of human frataxin by mitochondrial processing peptidase: Precursor and intermediate forms are cleaved at different rates",
abstract = "We showed previously that maturation of the human frataxin precursor (p-fxn) involves two cleavages by the mitochondrial processing peptidase (MPP). This observation was not confirmed by another group, however, who reported only one cleavage. Here, we demonstrate conclusively that MPP cleaves p-fxn in two sequential steps, yielding a 18,826-Da intermediate (i-fxn) and a 17,255-Da mature (m-fxn) form, the latter corresponding to endogenous frataxin in human tissues. The two cleavages occur between residues 41-42 and 55-56, and both match the MPP consensus sequence RX ↓ (X/S). Recombinant rat and yeast MPP catalyze the p → i step 4 and 40 times faster, respectively, than the i → m step. In isolated rat mitochondria, p-fxn undergoes a sequence of cleavages, p → i → m → d1 → d2, with d1 and d2 representing two C-terminal fragments of m-fxn produced by an unknown protease. The i → m step is limiting, and the overall rate of p → i → m does not exceed the rate of m → d1 → d2, such that the levels of m-fxn do not change during incubations as long as 3 h. Inhibition of the i → m step by a disease-causing frataxin mutation (W173G) leads to nonspecific degradation of i-fxn. Thus, the second of the two processing steps catalyzed by MPP limits the levels of mature frataxin within mitochondria.",
author = "Patrizia Cavadini and Jiri Adamec and Franco Taroni and Oleksandr Gakh and Grazia Isaya",
year = "2000",
month = "12",
day = "29",
doi = "10.1074/jbc.M006539200",
language = "English (US)",
volume = "275",
pages = "41469--41475",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "52",

}

TY - JOUR

T1 - Two-step processing of human frataxin by mitochondrial processing peptidase

T2 - Precursor and intermediate forms are cleaved at different rates

AU - Cavadini, Patrizia

AU - Adamec, Jiri

AU - Taroni, Franco

AU - Gakh, Oleksandr

AU - Isaya, Grazia

PY - 2000/12/29

Y1 - 2000/12/29

N2 - We showed previously that maturation of the human frataxin precursor (p-fxn) involves two cleavages by the mitochondrial processing peptidase (MPP). This observation was not confirmed by another group, however, who reported only one cleavage. Here, we demonstrate conclusively that MPP cleaves p-fxn in two sequential steps, yielding a 18,826-Da intermediate (i-fxn) and a 17,255-Da mature (m-fxn) form, the latter corresponding to endogenous frataxin in human tissues. The two cleavages occur between residues 41-42 and 55-56, and both match the MPP consensus sequence RX ↓ (X/S). Recombinant rat and yeast MPP catalyze the p → i step 4 and 40 times faster, respectively, than the i → m step. In isolated rat mitochondria, p-fxn undergoes a sequence of cleavages, p → i → m → d1 → d2, with d1 and d2 representing two C-terminal fragments of m-fxn produced by an unknown protease. The i → m step is limiting, and the overall rate of p → i → m does not exceed the rate of m → d1 → d2, such that the levels of m-fxn do not change during incubations as long as 3 h. Inhibition of the i → m step by a disease-causing frataxin mutation (W173G) leads to nonspecific degradation of i-fxn. Thus, the second of the two processing steps catalyzed by MPP limits the levels of mature frataxin within mitochondria.

AB - We showed previously that maturation of the human frataxin precursor (p-fxn) involves two cleavages by the mitochondrial processing peptidase (MPP). This observation was not confirmed by another group, however, who reported only one cleavage. Here, we demonstrate conclusively that MPP cleaves p-fxn in two sequential steps, yielding a 18,826-Da intermediate (i-fxn) and a 17,255-Da mature (m-fxn) form, the latter corresponding to endogenous frataxin in human tissues. The two cleavages occur between residues 41-42 and 55-56, and both match the MPP consensus sequence RX ↓ (X/S). Recombinant rat and yeast MPP catalyze the p → i step 4 and 40 times faster, respectively, than the i → m step. In isolated rat mitochondria, p-fxn undergoes a sequence of cleavages, p → i → m → d1 → d2, with d1 and d2 representing two C-terminal fragments of m-fxn produced by an unknown protease. The i → m step is limiting, and the overall rate of p → i → m does not exceed the rate of m → d1 → d2, such that the levels of m-fxn do not change during incubations as long as 3 h. Inhibition of the i → m step by a disease-causing frataxin mutation (W173G) leads to nonspecific degradation of i-fxn. Thus, the second of the two processing steps catalyzed by MPP limits the levels of mature frataxin within mitochondria.

UR - http://www.scopus.com/inward/record.url?scp=0034731447&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034731447&partnerID=8YFLogxK

U2 - 10.1074/jbc.M006539200

DO - 10.1074/jbc.M006539200

M3 - Article

C2 - 11020385

AN - SCOPUS:0034731447

VL - 275

SP - 41469

EP - 41475

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 52

ER -