Assay procedures for ascorbic acid 2-sulfate sulfohydrolase

Stevens, Richard L; Fluharty, Arvan L.; Shapiro, Stanley S.; Miller, Ruby T.; Davis, Laurel L.; Kihara, Hayato

doi:10.1016/0003-2697(77)90374-8

Cited by 8 publications

(2 citation statements)

References 14 publications

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“…The AA2S assay was modified from Stevens et al [12]. For partially puri fied enzymes, 40 pi of enzyme was combined with 40 pi 20 mM AA2S and 10 mM EDTA in 0.1 M Na acetate, pH 4.5.…”

Section: Methodsmentioning

confidence: 99%

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Isolation and Characterization of Rat Hepatic Ascorbic Acid-2-Sulfatases

Thompson¹,

Daniel²

1987

Enzyme

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Ascorbic acid-2-sulfatase was isolated from rat liver by a multistep procedure. DEAE Sephacel ion-exchange chromatography resolved crude ascorbic acid-2-sulfatase into cationic and anionic fractions. These fractions were purified 75- and 230-fold, respectively. The comparative biochemical properties suggest that arylsulfatase B is responsible for the cationic ascorbic acid-2-sulfatase activity, while arylsulfatase A appears to be responsible for the anionic ascorbic acid-2-sulfatase activity. Partially purified arylsulfatase A hydrolyzed ascorbic acid-2-sulfate at 4% the rate of p-nitrocatechol sulfate hydrolysis, while arylsulfatase B hydrolyzed ascorbic acid-2-sulfate at 0.6% the p-nitrocatechol sulfate rate.

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“…The AA2S assay was modified from Stevens et al [12]. For partially puri fied enzymes, 40 pi of enzyme was combined with 40 pi 20 mM AA2S and 10 mM EDTA in 0.1 M Na acetate, pH 4.5.…”

Section: Methodsmentioning

confidence: 99%

“…After incubation the assay mixture was cen trifuged for 15 min in an Eppendorf microfuge at 40 C. The supernatant was then mixed with 0.075 mM DCIP and the decrease in absorbance fol lowed at 600 nm. An extinction coefficient of 18,000 was used for the calculation of enzyme activity [12],…”

Section: Methodsmentioning

confidence: 99%

Isolation and Characterization of Rat Hepatic Ascorbic Acid-2-Sulfatases

Thompson¹,

Daniel²

1987

Enzyme

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Hydrocortisone regulates arylsulfatase A (cerebroside-3-sulfate-3-sulfohydrolase) by decreasing the quantity of the enzyme in cultures of cells dissociated from embryonic mouse cerebra

Marcelo

Pieringer

1990

Neurochem Res

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Previous work from our laboratory (Biochem. J. 219:689-697 (1984] had shown that hydrocortisone stimulated the net accumulation of the myelin-specific sulfolipid in cultures of cells dissociated from embryonic mouse cerebra. This accumulation caused by hydrocortisone was shown to be due to a decrease of sulfolipid degradation by arylsulfatase A (ASA) and not due to a stimulation of its synthesis by a sulfotransferase. Both ASA activity and the turnover of sulfolipid were decreased by hydrocortisone to 60-62% of untreated cells. In current work the same decrease in enzyme activity was obtained and enzyme linked immunosorbent assays demonstrate that hydrocortisone decreased the number of ASA protein molecules to 61% of untreated cells [(-)hydrocortisone: 0.31 +/- 0.06 ng ASA/microgram protein; (+)hydrocortisone: 0.18 +/- 0.04 ng ASA/microgram protein]. This decrease in the number of ASA molecules correlates well with the decrease in both the enzyme activity and the sulfolipid turnover, which suggests that the major mode of inhibition of ASA activity by hydrocortisone involves a decrease in the concentration of ASA in the cells rather than some other mechanism of inhibition.

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Specific determination of arylsulfatase A activity

1986

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Arylsulfatase activities in biological materials are too low to be detected by the methods available hitherto. A sensitive and specific assay method for arylsulfatase A (AS-A) has been developed in the present study. Ascorbate-2-sulfate is known to be a specific natural substrate of AS-A; the ascorbic acid liberated by the action of AS-A was quantitatively determined using HPLC equipped with an amperometric detector. The method was used to analyze the activity of AS-A in biological materials.

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Assay procedures for ascorbic acid 2-sulfate sulfohydrolase

Cited by 8 publications

References 14 publications

Isolation and Characterization of Rat Hepatic Ascorbic Acid-2-Sulfatases

Isolation and Characterization of Rat Hepatic Ascorbic Acid-2-Sulfatases

Hydrocortisone regulates arylsulfatase A (cerebroside-3-sulfate-3-sulfohydrolase) by decreasing the quantity of the enzyme in cultures of cells dissociated from embryonic mouse cerebra

Specific determination of arylsulfatase A activity

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