Proteolytic degradation of tyrosine nitrated proteins.

Tyrosine nitration is a covalent posttranslational protein modification that has been detected under several pathological conditions. This study reports that nitrated proteins are degraded by chymotrypsin and that protein nitration enhances susceptibility to degradation by the proteasome. Chymotrypsin cleaved the peptide bond between nitrated-tyrosine 108 and serine 109 in bovine Cu,Zn superoxide dismutase. However, the rate of chymotryptic cleavage of nitrated peptides was considerably slower than control. In contrast, nitrated bovine Cu,Zn superoxide dismutase was degraded at a rate 1. 8-fold faster than that of control by a gradient-purified 20S/26S proteasome fraction from bovine retina. Exposure of PC12 cells to a nitrating agent resulted in the nitration of tyrosine hydroxylase and a 58 +/- 12.5% decline in the steady-state levels of the protein 4 h after nitration. The steady-state levels of tyrosine hydroxylase were restored by selective inhibition of the proteasome activity with lactacystin. These data indicate that nitration of tyrosine residue(s) in proteins is sufficient to induce an accelerated degradation of the modified proteins by the proteasome and that the proteasome may be critical for the removal of nitrated proteins in vivo.

[1]  Alexander Varshavsky,et al.  The ubiquitin system. , 1998, Annual review of biochemistry.

[2]  F. Hartl,et al.  Responses to Peroxynitrite in Yeast: Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) as a Sensitive Intracellular Target for Nitration and Enhancement of Chaperone Expression and Ubiquitination , 2000, Biological chemistry.

[3]  M. Scheffner,et al.  Activation of the Cell Death Program by Nitric Oxide Involves Inhibition of the Proteasome* , 1999, The Journal of Biological Chemistry.

[4]  C. Schöneich,et al.  Protein modification during biological aging: selective tyrosine nitration of the SERCA2a isoform of the sarcoplasmic reticulum Ca2+-ATPase in skeletal muscle. , 1999, The Biochemical journal.

[5]  A. Haas,et al.  Neurite Outgrowth in PC12 Cells , 1999, The Journal of Biological Chemistry.

[6]  R. Radi,et al.  Peroxynitrite inhibits T lymphocyte activation and proliferation by promoting impairment of tyrosine phosphorylation and peroxynitrite-driven apoptotic death. , 1999, Journal of immunology.

[7]  J. Rychik,et al.  Changes in oxygenation with inhaled nitric oxide in severe bronchopulmonary dysplasia. , 1999, Pediatrics.

[8]  V. Ullrich,et al.  Interleukin 1beta decreases prostacyclin synthase activity in rat mesangial cells via endogenous peroxynitrite formation. , 1998, The Biochemical journal.

[9]  T. Reinheckel,et al.  Comparative resistance of the 20S and 26S proteasome to oxidative stress. , 1998, The Biochemical journal.

[10]  H. Ischiropoulos.  Biological tyrosine nitration: a pathophysiological function of nitric oxide and reactive oxygen species. , 1998, Archives of biochemistry and biophysics.

[11]  M. Strong,et al.  Nitration of the low molecular weight neurofilament is equivalent in sporadic amyotrophic lateral sclerosis and control cervical spinal cord. , 1998, Biochemical and biophysical research communications.

[12]  S. Goto,et al.  Age-related changes in the 20S and 26S proteasome activities in the liver of male F344 rats , 1998, Mechanisms of Ageing and Development.

[13]  K. Davies,et al.  Peroxynitrite Increases the Degradation of Aconitase and Other Cellular Proteins by Proteasome* , 1998, The Journal of Biological Chemistry.

[14]  J. Blumberg,et al.  Regulation of Ubiquitin-conjugating Enzymes by Glutathione Following Oxidative Stress* , 1997, The Journal of Biological Chemistry.

[15]  C. Pickart.  Targeting of substrates to the 26S proteasome , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[16]  T. Reinheckel,et al.  Degradation of oxidized proteins in mammalian cells , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[17]  A. Gow,et al.  Effects of peroxynitrite‐induced protein modifications on tyrosine phosphorylation and degradation , 1996, FEBS letters.

[18]  K Tanaka,et al.  Structure and functions of the 20S and 26S proteasomes. , 1996, Annual review of biochemistry.

[19]  S. Matalon,et al.  Quantitation of nitrotyrosine levels in lung sections of patients and animals with acute lung injury. , 1994, The Journal of clinical investigation.

[20]  M. J. van der Woerd,et al.  Crystal structure of peroxynitrite-modified bovine Cu,Zn superoxide dismutase. , 1992, Archives of biochemistry and biophysics.

[21]  J S Beckman,et al.  Kinetics of superoxide dismutase- and iron-catalyzed nitration of phenolics by peroxynitrite. , 1992, Archives of biochemistry and biophysics.

[22]  J S Beckman,et al.  Peroxynitrite-mediated tyrosine nitration catalyzed by superoxide dismutase. , 1992, Archives of biochemistry and biophysics.

[23]  K. Davies,et al.  Protein degradation as an index of oxidative stress. , 1990, Methods in enzymology.

[24]  W. Huttner.  Protein tyrosine sulfation , 1987 .