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.
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U2 - 10.1074/jbc.M006539200
DO - 10.1074/jbc.M006539200
M3 - Article
C2 - 11020385
AN - SCOPUS:0034731447
SN - 0021-9258
VL - 275
SP - 41469
EP - 41475
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 52
ER -