Cerebroside sulfatase determination in cultured human fibroblasts

Porter, Myna T.; Fluharty, Arvan L.; Flor, Syliva D. De La; Kihara, Hayato

doi:10.1016/0005-2744(72)90178-7

Cited by 65 publications

(17 citation statements)

References 19 publications

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“…The optimal TDC-promoted sulfohydrolase rate was approximately fourfold greater than the CSS activator facilitated system (data not shown). This bile salt shows a sharp optimum for the CSS reaction at about 1.25 mg/ml, just below its critical micellar concentration [7]. A similar TDC concentration optimum was seen for the myelin-bound substrate.…”

Section: Resultsmentioning

confidence: 59%

“…They succeeded in identifying a low-molecular-weight, heatstable auxiliary protein which allowed ARS-A to act on CS, but did not affect the ability of the enzyme to act on water-soluble synthetic substrates. It was later learned that certain bile salts were also capable of facilitating CS breakdown by ARS-A [6,7]. When it was established that CS was not truly soluble in aqueous buffers, but existed as large aggregates even in optically clear CS suspensions [8], the activator role was considered to be that of lipid dispersal.…”

Section: Introductionmentioning

confidence: 99%

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Activator-Dependent Hydrolysis of Myelin Cerebroside Sulfate by Arylsulfatase A

Louis¹,

Fluharty²

1991

Dev Neurosci

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A purified myelin preparation containing [³⁵S]-labeled cerebroside sulfate (CS) was biosynthesized in developing rat brain and tested as a model of a physiological substrate for CS hydrolysis by arylsulfatase A. Particular attention was directed to the involvement of the CS sulfatase activator protein in facilitating the catabolic process. Although arylsulfatase A alone was incapable of desulfating CS in either purified CS suspensions or the physiological membrane, activator-induced hydrolysis of myelin CS exhibited concentration dependency, pH optimum, and relative insensitivity to salts in a manner similar to purified lipid suspensions. Exogenous protein demonstrated concentration-dependent inhibition. Slower rates of hydrolysis observed for the myelin membrane substrate are proposed to be a consequence of myelin membrane configuration and competition for activator by other lipoidal constituents.

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Section: Resultsmentioning

confidence: 59%

Section: Introductionmentioning

confidence: 99%

Activator-Dependent Hydrolysis of Myelin Cerebroside Sulfate by Arylsulfatase A

Louis¹,

Fluharty²

1991

Dev Neurosci

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“…4, B and C). Conventional ASA activity assays circumvent the need for Sap B by using either a detergent which mimics the mobilization function of Sap B (9,34) or by using water-soluble substrates like the sulfate ester pNCS (21,30). In both experimental set-ups the reaction rate is determined by the catalytic rate of ASA (Fig.…”

Section: Discussionmentioning

confidence: 99%

Saposin B-dependent Reconstitution of Arylsulfatase A Activity in Vitro and in Cell Culture Models of Metachromatic Leukodystrophy

Matzner

Breiden

Schwarzmann

et al. 2009

Journal of Biological Chemistry

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“…The fibroblast loading test was carried out as described by Kihara et al (1980), using [35S] labeled cerebroside sulfate prepared in rats (Fluharty et al 1974). Cerebroside sulfatase activity was determined by type I activation (Porter et al 1972) and by type I1 activation (Fluharty et al 1980). Activity toward 4-methylumbelliferone sulfate was assayed as described by Fluharty & Edmond (1978).…”

Section: Methodsmentioning

confidence: 99%

“…A requirement for a protein activator for cerebroside sulfatase activity in cell-free assays was discovered by Mehl & Jatzkewitz (1964). The cerebroside sulfatase activator function can also be fulfilled by low concentrations of sodium taurodeoxycholate (Porter et al 1972) or sodium cholate (Percy et al 1972). We have designated this mode of activation as type I activation to differentiate it from type I1 activation whioh is promoted by high concentrations of cholate.…”

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confidence: 99%

Metachromatic leukodystrophy caused by a partial cerebroside sulfatase defect

Kihara¹,

Fluharty²,

O’Brien

et al. 1982

Clinical Genetics

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A patient with neuropathy and myopathy since infancy but whose neuropathy had been stable for a number of years showed a profound deficiency of arylsulfatase A in leukocytes and urine. Urine contained material that stained metachromatically and cochromatographed with cerebroside sulfate. In contrast, cultured fibroblasts contained about 10–20 % of normal arylsulfatase A with properties identical to properties of normal fibroblast enzyme, except that it showed no cerebroside sulfatase activity. Growing fibroblasts in the cerebroside sulfate loading test had an attenuated rate of sulfatide hydrolysis. A reexamination of the cerebroside sulfatase reaction revealed that while only limited hydrolysis occurred with low concentrations of taurodeoxycholate or chelate (type I activation), significant hydrolysis of the natural substrate did take place with high concentrations of cholate (type II activation). This suggests that there is a partial cerebroside sulfatase defect in this atypical form of metachromatic leukodystrophy.

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Cerebroside sulfatase determination in cultured human fibroblasts

Cited by 65 publications

References 19 publications

Activator-Dependent Hydrolysis of Myelin Cerebroside Sulfate by Arylsulfatase A

Activator-Dependent Hydrolysis of Myelin Cerebroside Sulfate by Arylsulfatase A

Saposin B-dependent Reconstitution of Arylsulfatase A Activity in Vitro and in Cell Culture Models of Metachromatic Leukodystrophy

Metachromatic leukodystrophy caused by a partial cerebroside sulfatase defect

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