Stearoyl[1-14C]sulfogalactosylsphingosine ([14C]sulfatide) as substrate for cerebroside sulfatase assay

Dubois, G; Zalc, B.; Saux, Françoise Le; Baumann, Nicole

doi:10.1016/0003-2697(80)90159-1

Cited by 48 publications

(24 citation statements)

References 14 publications

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“…The substrate is of very high radiopurity (99.6%) and is relatively simple to prepare according to a published procedure (23). Previous studies have demonstrated that CS could be incorporated into cultured skin fibroblasts, and that it could induce the formation of metachromatic inclusions in cells from MLD patients (24).…”

Section: Discussionmentioning

confidence: 99%

Diagnosis of Metachromatic Leukodystrophy, Krabbe Disease, and Farber Disease after Uptake of Fatty Acid-labeled Cerebroside Sulfate into Cultured Skin Fibroblasts

Kudoh¹,

Wenger²

1982

J. Clin. Invest.

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A B S T R A C T ['4C]Stearic acid-labeled cerebroside sulfate (CS) was presented to cultured skin fibroblasts in the media. After endocytosis into control cells 86% was readily metabolized to galactosylceramide, ceramide, and stearic acid, which was reutilized in the synthesis of the major lipids found in cultured fibroblasts. Uptake and metabolism of the ['4C]CS into cells from typical and atypical patients and carriers of metachromatic leukodystrophy (MLD), Krabbe disease, and Farber disease were observed. Cells from patients with late infantile MLD could not metabolize the CS at all, while cells from an adult MLD patient and from a variant MLD patient could metabolize -40 and 15%, respectively, of the CS taken up. These results are in contrast to the in vitro results that demonstrated a severe deficiency of arylsulfatase A in the late infantile and adult patient and a partial deficiency (21-27% of controls) in the variant MLD patient. Patients with Krabbe disease could metabolize nearly 40% of the galactosylceramide produced in the lysosomes from the CS. This is in contrast to the near zero activity for galactosylceramidase measured in vitro. Carriers of Krabbe disease with galactosylceramidase activity near half normal in vitro and those with under 10% of normal activity were found to metabolize galactosylceramide in cells significantly slower than controls. This provides a method for differentiating affected patients from carriers with low enzyme activity in vitro. Cells from patients with Farber disease could catabolize only '-15% of the ceramide produced from

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Section: Discussionmentioning

confidence: 99%

Diagnosis of Metachromatic Leukodystrophy, Krabbe Disease, and Farber Disease after Uptake of Fatty Acid-labeled Cerebroside Sulfate into Cultured Skin Fibroblasts

Kudoh¹,

Wenger²

1982

J. Clin. Invest.

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“…None of the ' 4C-, 3H-, or 35S-labeled substrates employed for this purpose (Dubois et a!., 1980;Raghavan et a!., 1981;Porter et al, 1971) are available commercially and must, therefore, be prepared. The 5S-sulfate labeled material (C35S) is the simplest to prepare and use, but its short Fluorescent Substrate for Arylsulfatase 115 half-life (87 d) necessitates frequent preparation.…”

Section: Introductionmentioning

confidence: 99%

Synthesis and characterization of NBD-PS: A fluorescent analog of cerebroside sulfate for diagnosis of arylsulfatase a deficiency disorders

Louis

Widen

Tsay

et al. 1991

Molecular and Chemical Neuropathology

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N-[7-Nitrobenz-2-oxa-1,3-diazol-4-yl]psychosine sulfate (NBD-PS), a fluorescent analog of cerebroside sulfate (CS), was synthesized and tested as an alternative to the radiolabeled forms of CS used for assaying arylsulfatase A (ASA) in its physiological role as a cerebroside sulfate sulfohydrolase. NBD-PS simulates the natural substrate for ASA. Protocols have been developed for its use in differentiating low enzyme activities in diagnostic samples. Hydrolysis of NBD-PS is specific for ASA and optimal assay parameters were identical to those determined for CS. Differentiations between each of the major phenotypes for ASA activity were possible in the set of samples tested. One particular advantage was the ability to discriminate between individuals exhibiting arylsulfatase A pseudodeficiency and the truly deficient individuals with metachromatic leukodystrophy. Differential diagnosis was possible with fibroblast extracts by an assay that is more sensitive than procedures employing radioisotopes. Reaction products may be analyzed quantitatively by HPLC, or semiquantitatively with TLC. NBD-PS provides a simpler, safer, and more cost-effective means of performing natural substrate enzyme assays for ASA. Phenotyping with the fluorescence assay is an effective alternative to the laborious radioactive CS preparations and tissue culture loading studies that have previously been necessary.

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“…Sulfatide was N-deacylated and reacylated with stearoyl chloride [17,18] and the main products purifed again. The acetylated product was desulfated with p-toluenesulfonic acid [20].…”

Section: Preparation Of Acceptor Moleculesmentioning

confidence: 99%

“…Galactocerebroside was submitted to alkaline Ndeacylation and reacylation with stearoyl chloride [17,18] and the main products purifed again. Tritylation was made according to [19], the tritylated product was acetylated and detritylated [20].…”

Section: Preparation Of Acceptor Moleculesmentioning

confidence: 99%

Galactofuranosylated Galactocerebrosides, a New Drug Delivery System for Ceramides to Colon

Iga¹,

Iga²

2008

TOOCJ

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Stearoyl[1-14C]sulfogalactosylsphingosine ([14C]sulfatide) as substrate for cerebroside sulfatase assay

Cited by 48 publications

References 14 publications

Diagnosis of Metachromatic Leukodystrophy, Krabbe Disease, and Farber Disease after Uptake of Fatty Acid-labeled Cerebroside Sulfate into Cultured Skin Fibroblasts

Diagnosis of Metachromatic Leukodystrophy, Krabbe Disease, and Farber Disease after Uptake of Fatty Acid-labeled Cerebroside Sulfate into Cultured Skin Fibroblasts

Synthesis and characterization of NBD-PS: A fluorescent analog of cerebroside sulfate for diagnosis of arylsulfatase a deficiency disorders

Galactofuranosylated Galactocerebrosides, a New Drug Delivery System for Ceramides to Colon

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