Aspartoacylase is a regulated nuclear-cytoplasmic enzyme.

Abstract

Mutations in the gene for aspartoacylase (ASPA), which catalyzes deacetylation of N-acetyl-L-aspartate in the central nervous system (CNS), result in Canavan Disease, a fatal dysmyelinating disease. Consistent with its role in supplying acetate for myelin lipid synthesis, ASPA is thought to be cytoplasmic. Here we describe the occurrence of ASPA within nuclei of rat brain and kidney, and in cultured rodent oligodendrocytes. Immunohistochemistry showed cytoplasmic and nuclear ASPA staining, the specificity of which was demonstrated by its absence from tissues of the Tremor rat, an ASPA-null mutant. Subcellular fractionation analysis revealed low enzyme activity against NAA in nuclear fractions from normal rats. Whereas two recent reports have indicated that ASPA exists as a dimer, size-exclusion chromatography of subcellular fractions showed ASPA is an active monomer in both subcellular fractions. Western blotting detected ASPA as a single 38 kD band. Because ASPA is small enough to passively diffuse into the nucleus, we constructed, expressed, and detected in COS-7 cells a green fluorescent protein-human ASPA (GFP-hASPA) fusion protein larger than the permissible size for the nuclear pore complex. GFP-hASPA was enzymatically active and showed mixed nuclear-cytoplasmic distribution. We conclude that ASPA is a regulated nuclear-cytoplasmic protein that may have distinct functional roles in the two cellular compartments.

• Authors: Jeremy R Hershfield, Chikkathur N Madhavarao, John R Moffett, Joyce A Benjamins, James Y Garbern, Aryan Namboodiri
• Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology (FASEB J) Vol. 20 Issue 12 Pg. 2139-41 (Oct 2006) ISSN: 1530-6860 [Electronic] United States
• PMID: 16935940 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Green Fluorescent Proteins
• Amidohydrolases
• aspartoacylase

Topics

• Active Transport, Cell Nucleus
• Amidohydrolases (analysis, genetics, isolation & purification)
• Animals
• Brain (enzymology)
• Cell Fractionation
• Cell Nucleus (enzymology)
• Cytoplasm (enzymology)
• Green Fluorescent Proteins (genetics)
• Humans
• Kidney (enzymology)
• Rats
• Transfection