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

Varda Barash; Daphna Flhor; Bilha Morag; Avihu Boneh; Orly N. Elpeleg; Chaim Gilon

doi:10.1016/0009-8981(91)90368-M

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

Varda Barash^*, Daphna Flhor, Bilha Morag, Avihu Boneh, Orly N. Elpeleg, Chaim Gilon

^*Corresponding author for this work

Institute of Chemistry

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

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.

Original language	English
Pages (from-to)	175-181
Number of pages	7
Journal	Clinica Chimica Acta
Volume	201
Issue number	3
DOIs	https://doi.org/10.1016/0009-8981(91)90368-M
State	Published - 30 Sep 1991

Keywords

Aspartoacylase
Canavan's disease
Leukodystrophy
Spongy degeneration of brain

Access to Document

10.1016/0009-8981(91)90368-M

Cite this

@article{1a47ae6839f24683aa79d62f7d643caf,

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

abstract = "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.",

keywords = "Aspartoacylase, Canavan's disease, Leukodystrophy, Spongy degeneration of brain",

author = "Varda Barash and Daphna Flhor and Bilha Morag and Avihu Boneh and Elpeleg, {Orly N.} and Chaim Gilon",

year = "1991",

month = sep,

day = "30",

doi = "10.1016/0009-8981(91)90368-M",

language = "אנגלית",

volume = "201",

pages = "175--181",

journal = "Clinica Chimica Acta",

issn = "0009-8981",

publisher = "Elsevier B.V.",

number = "3",

}

TY - JOUR

T1 - A radiometric assay for aspartoacylase activity in human fibroblasts

T2 - application for the diagnosis of Canavan's disease

AU - Barash, Varda

AU - Flhor, Daphna

AU - Morag, Bilha

AU - Boneh, Avihu

AU - Elpeleg, Orly N.

AU - Gilon, Chaim

PY - 1991/9/30

Y1 - 1991/9/30

N2 - 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.

AB - 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.

KW - Aspartoacylase

KW - Canavan's disease

KW - Leukodystrophy

KW - Spongy degeneration of brain

UR - http://www.scopus.com/inward/record.url?scp=0026040840&partnerID=8YFLogxK

U2 - 10.1016/0009-8981(91)90368-M

DO - 10.1016/0009-8981(91)90368-M

M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???

C2 - 1756590

AN - SCOPUS:0026040840

SN - 0009-8981

VL - 201

SP - 175

EP - 181

JO - Clinica Chimica Acta

JF - Clinica Chimica Acta

IS - 3

ER -

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

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this