Tyrosine nitration impairs mammalian aldolase A activity

Mol Cell Proteomics. 2004 Jun;3(6):548-57. doi: 10.1074/mcp.M300141-MCP200. Epub 2004 Feb 20.


Protein tyrosine nitration increases in vivo as a result of oxidative stress and is elevated in numerous inflammatory-associated diseases. Mammalian fructose-1,6-bisphosphate aldolases are tyrosine nitrated in lung epithelial cells and liver, as well as in retina under different inflammatory conditions. Using two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, we now show that aldolase A is nitrated in human skin fibroblasts. To reveal the consequences of tyrosine nitration, we studied the impact of peroxynitrite on the glycolytic functions of aldolase A. A peroxynitrite concentration-dependent decrease in fructose-1,6-bisphosphate cleavage activity was observed with a concomitant increase in nitrotyrosine immunoreactivity. Both V(max) and the K(m) for fructose-1,6-bisphosphate decreased after incubation with peroxynitrite. Aldolase nitrotyrosine immunoreactivity diminished following carboxypeptidase Y digestion, demonstrating that tyrosine residues in the carboxyl-terminal region of aldolase are major targets of nitration. Aldolase A contains a carboxyl-terminal tyrosine residue, Tyr(363), that is critical for its catalytic activity. Indeed, tandem mass spectrometric analysis of trypsin-digested aldolase showed that Tyr(363) is the most susceptible to nitration, with a modification of Tyr(342) occurring only after nitration of Tyr(363). These tyrosine nitrations likely result in altered interactions between the carboxyl-terminal region and enzyme substrate or reaction intermediates causing the decline in activity. The results suggest that tyrosine nitration of aldolase A can contribute to an impaired cellular glycolytic activity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Electrophoresis, Gel, Two-Dimensional
  • Fibroblasts / enzymology
  • Fructose-Bisphosphate Aldolase / metabolism*
  • Humans
  • Nitrosation
  • Skin / enzymology*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Tyrosine / analogs & derivatives*
  • Tyrosine / chemistry
  • Tyrosine / metabolism*


  • 3-nitrotyrosine
  • Tyrosine
  • Fructose-Bisphosphate Aldolase