A new analytical bench assay for the determination of arylsulfatase a activity toward galactosyl-3-sulfate ceramide

Implication for metachromatic leukodystrophy diagnosis

Francesco Morena, Ilaria Di Girolamo, Carla Emiliani, Angela Gritti, Alessandra Biffi, Sabata Martino

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Here, we present the design and validation of a new assay for the diagnosis of metachromatic leukodystrophy. The method is highly specific, simple, reproducible, and straightforward. In our spectrophotometric method, the determination of arylsulfatase A (ARSA) activity toward the natural substrate, galactosyl-3-sulfate ceramide (or sulfatide), is performed using neat sulfatide without chemical modification. This confers to the assay high analytical specificity. The hydrolyzed sulfatide is monitored upon inclusion of the colorimetric reagent Azure A. The nonhydrolyzed sulfatide-Azure A is recovered and measured at a wavelength of λ = 650 nm. Thus, ARSA activity toward the sulfatide is obtained by subtracting the nonhydrolyzed sulfatide from the total sulfatide used in the enzyme reaction (sulfatide-Azure A present in a parallel assay performed in the absence of ARSA). Within a clinical context, our method definitely discriminated between healthy subject samples and metachromatic leukodystrophy patient samples, and, therefore, it is suitable for diagnostic applications and for monitoring the efficacy of therapeutic treatments in patients or animal models.

Original languageEnglish
Pages (from-to)473-481
Number of pages9
JournalAnalytical Chemistry
Volume86
Issue number1
DOIs
Publication statusPublished - Jan 7 2014

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Arylsulfatases
Sulfoglycosphingolipids
Ceramides
Sulfates
Assays
Cerebroside-Sulfatase
Chemical modification
Animals
Wavelength
Monitoring

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

A new analytical bench assay for the determination of arylsulfatase a activity toward galactosyl-3-sulfate ceramide : Implication for metachromatic leukodystrophy diagnosis. / Morena, Francesco; Di Girolamo, Ilaria; Emiliani, Carla; Gritti, Angela; Biffi, Alessandra; Martino, Sabata.

In: Analytical Chemistry, Vol. 86, No. 1, 07.01.2014, p. 473-481.

Research output: Contribution to journalArticle

Morena, Francesco ; Di Girolamo, Ilaria ; Emiliani, Carla ; Gritti, Angela ; Biffi, Alessandra ; Martino, Sabata. / A new analytical bench assay for the determination of arylsulfatase a activity toward galactosyl-3-sulfate ceramide : Implication for metachromatic leukodystrophy diagnosis. In: Analytical Chemistry. 2014 ; Vol. 86, No. 1. pp. 473-481.
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