Purification, characterization, and localization of aspartoacylase from bovine brain

Rajinder Kaul, Jose Casanova, Anne B. Johnson, Peter Tang, Reuben Matalon

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

Canavan disease, an autosomal recessive disorder, is characterized biochemically by N-acetylaspartic aciduria and aspartoacylase (N-acyl-L-aspartate amidohydrolase; EC 3.5.1.15) deficiency. However, the role of aspartoacylase and N-acetylaspartic acid in brain metabolism is unknown. Aspartoacylase has been purified to apparent homogeneity with a specific activity of ∼ 19,000-20,000 nmol of aspartate released/mg of protein. The native enzyme is a 58-kDa monomer. The purified aspartoacylase activity is enhanced by divalent cations, nonionic detergents, and dithiothreitol. Low levels of dithiothreitol or β-mercaptoethanol are required for enzyme stability. Aspartoacylase has a Km of 8.5 × 10-4 M and a Vmax of 43,000 nmol/min/mg of protein. Inhibition of aspartoacylase by glycyl-L-aspartate and amino derivatives of D-aspartic acid suggests that the carbon backbone of the substrate is primarily involved in its interaction with the active site and that a blocked amino group is essential for the catalytic activity of aspartoacylase. Biochemical and immunocytochemical studies revealed that aspartoacylase is localized to white matter, whereas the N-acetylaspartic acid concentration is threefold higher in gray matter than in white matter. Our studies so far indicate that aspartoacylase is conserved across species during evolution and suggest a significant role for aspartoacylase and N-acetylaspartic acid in normal brain biology.

Original languageEnglish (US)
Pages (from-to)129-135
Number of pages7
JournalJournal of Neurochemistry
Volume56
Issue number1
StatePublished - 1991
Externally publishedYes

Fingerprint

Purification
Brain
Dithiothreitol
Aspartic Acid
Canavan Disease
aspartoacylase
D-Aspartic Acid
Enzyme Stability
Mercaptoethanol
Divalent Cations
Enzymes
Metabolism
Detergents
Catalyst activity
Catalytic Domain
Proteins
Carbon
Monomers
Derivatives
Substrates

Keywords

  • Aminoacylase II
  • Canavan disease
  • Myelin
  • N-acetylaspartic acid
  • Spongy degeneration

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Purification, characterization, and localization of aspartoacylase from bovine brain. / Kaul, Rajinder; Casanova, Jose; Johnson, Anne B.; Tang, Peter; Matalon, Reuben.

In: Journal of Neurochemistry, Vol. 56, No. 1, 1991, p. 129-135.

Research output: Contribution to journalArticle

Kaul, R, Casanova, J, Johnson, AB, Tang, P & Matalon, R 1991, 'Purification, characterization, and localization of aspartoacylase from bovine brain', Journal of Neurochemistry, vol. 56, no. 1, pp. 129-135.
Kaul, Rajinder ; Casanova, Jose ; Johnson, Anne B. ; Tang, Peter ; Matalon, Reuben. / Purification, characterization, and localization of aspartoacylase from bovine brain. In: Journal of Neurochemistry. 1991 ; Vol. 56, No. 1. pp. 129-135.
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