A radiometric assay for aspartoacylase activity in human fibroblasts: application for the diagnosis of Canavan's disease

Clin Chim Acta. 1991 Sep 30;201(3):175-81. doi: 10.1016/0009-8981(91)90368-m.


A new sensitive method for measuring aspartoacylase activity in human skin fibroblasts using [3H]N-acetyl-L-aspartic acid (NAA) is described. Optimal assay conditions and kinetic parameters for enzyme activity were determined. The enzyme was found to have maximal activity at pH 8.5, and the Michaelis constant for the substrate N-acetylaspartate was 1.8-2.0 mmol/l. Aspartoacylase activity in control cultured human fibroblasts was 9.2 +/- 1.8 nmol/h per mg protein, compared with 1.1 +/- 0.2 in seven Canavan patients and 3.5 +/- 0.9 in four patients' parents. This method for determining aspartoacylase activity is advantageous to the previously described spectrophotometric method since it is rapid, more sensitive and has less nonspecific interference. It is possible that application of this technique to cultured ammniotic and chorionic villi cells may be used for prenatal diagnosis of Canavan's disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / analysis
  • Amidohydrolases / metabolism*
  • Aspartic Acid / analogs & derivatives
  • Aspartic Acid / metabolism
  • Cells, Cultured
  • Clinical Enzyme Tests
  • Diffuse Cerebral Sclerosis of Schilder / diagnosis*
  • Fibroblasts / cytology
  • Fibroblasts / enzymology*
  • Heterozygote
  • Humans
  • Kinetics
  • Nerve Degeneration
  • Radiometry
  • Spectrophotometry
  • Tritium


  • Acetates
  • Tritium
  • Aspartic Acid
  • N-acetylaspartate
  • Amidohydrolases
  • aspartoacylase