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J Biol Chem, Vol. 273, Issue 18, 10857-10862, May 1, 1998
§,
From the We report that exposure of aconitase to moderate
concentrations of peroxynitrite, 3-morpholinosydnonimine (SIN-1; a
superoxide- and nitric oxide-liberating substance), or hydrogen
peroxide, inhibits the enzyme and enhances susceptibility to
proteolytic digestion by the isolated 20 S proteasome. Exposure to more
severe levels of oxidative stress, from these same agents, causes
further inhibition of the enzymatic activity of aconitase but actually decreases its proteolytic breakdown by proteasome. It should be noted
that the superoxide and nitric oxide liberated by SIN-1 decomposition
react to form a steady flux of peroxynitrite.
S-Nitroso-N-acetylpenicillamine, a
compound that liberates nitric oxide alone, causes only a small loss of
aconitase activity (25% or less) and has no effect on the proteolytic
susceptibility of the enzyme. Proteasome also seems to be the main
protease in cell lysates that can degrade aconitase after it has been
oxidatively modified by exposure to peroxynitrite, SIN-1, or hydrogen
peroxide. Using cell lysates isolated from K562 cells treated for
several days with an antisense oligodeoxynucleotide to the initiation
codon region of the C2 subunit of proteasome (a treatment which
diminishes proteasome activity by 50-60%), the enhanced degradation
of moderately damaged aconitase was essentially abolished. Other model
proteins as well as complex mixtures of proteins, such as cell lysates,
also exhibit enhanced proteolytic susceptibility after moderate SIN-1
treatment. Therefore we conclude that peroxynitrite reacts readily with
proteins and that mild modification by peroxynitrite results in
selective recognition and degradation by proteasome.
Ethel Percy Andrus Gerontology Center,
University of Southern California, Los
Angeles, California 90089-0191, the § Clinics of Physical
Medicine and Rehabilitation, Medical Faculty (Charité),
Humboldt-University Berlin, D-10098 Berlin, Germany, and the
¶ Institute of Molecular Pharmacology, D-10315
Berlin, Germany
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