Glucocerebrosidase: reconstitution of activity from macromolecular components (<i>Short Communication)</i>

Ho, Mae‐Wan; O’Brien, John S.; Radin, Norman S.; Erickson, John S.

doi:10.1042/bj1310173

Cited by 82 publications

(24 citation statements)

References 11 publications

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“…As stated earlier, a low molecular weight glycoprotein from the spleen of Gaucher patients associates on in cubation with a membrane fragment from normal spleen to produce de novo glucocerebrosidase activity [34,35].…”

Section: Discussionmentioning

confidence: 99%

“…The two components responsible were isolated as a low molecular weight (20,000) glycoprotein (factor P) from the patients spleen, and a membrane frag ment (factor C) from controls. Incubation of the two together not only produced /j-glucosidase activity as measured with 4-methylumbelliferyl-/3-glucoside but also caused a more dramatic rise in specific glucocerbrosidase activity [35].…”

Section: ß-Glucosidasementioning

confidence: 99%

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Multiple Forms of Glycosidases in the Normal and Pathological States

Robinson¹

1974

Enzyme

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Multiple forms of α- and β-galactosidase, α- and β-glucosidase, α- and β-hexosaminidases, α-mannosidase, α-fucosidase, and α-iduronidase of human tissues are surveyed. Their relationship to known storage disorders of glycolipid, glycoprotein, and mucopolysaccharide metabolism are discussed. A sub-unit hypothesis is presented to explain the interrelationships of multiple forms of β-N-acetylhexosaminidases that may be of wider application to include other glycosidases.

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Section: Discussionmentioning

confidence: 99%

Section: ß-Glucosidasementioning

confidence: 99%

Multiple Forms of Glycosidases in the Normal and Pathological States

Robinson¹

1974

Enzyme

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“…It remains to be analyzed whether the activator proteins for the degradation of glucosylceramide by the lysosomal j-glucosidase, described by several authors [13,15,33,34], will stimulate the hydrolysis of liposomal glucosylceramide even better than anionic phospholipids in the detergent-free system. In contrast to other water-soluble lysosomal hydrolases, such as B-hexosaminidase A [35, 361 and sulphatide arylsulphatase [37,381, the highly purified watersoluble glucosylceramide fi-glucosidase does not need the presence of activator proteins to degrade the membranebound glucosylceramide, since it can interact directly with the amphiphilic substrate at the surface of the bilayer as shown here.…”

Section: -Muglcmentioning

confidence: 99%

Specificity of human glucosylceramide β‐glucosidase towards synthetic glucosylsphingolipids inserted into liposomes

Sarmientos

Schwarzmann

Sandhoff

1986

European Journal of Biochemistry

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The behaviour of highly purified glucosylceramide P-glucosidase (glucosylceramidase, EC 3.2.1.45) from human placenta [Furbish, F. S., Blair, H. E., Shiloach, J., Pentchev, P. G. & Brady, R.B. (1977) Proc. Natl Acad. Sci. USA 74, 3560-35631 was investigated in the absence of detergents with structurally modified glucosylceramides inserted into unilamellar liposomes. The reaction between the water-soluble enzyme and the liposomal substrates was significantly dependent on the structure of the lipophilic aglycon moiety of glycolipids: glucosyl-N-acetyl-sphingosines (D-erythro and L-threo) were better substrates than the corresponding glucosylceramides.The L-threo derivatives were poorer substrates with higher apparent K, values than the corresponding D-erythro derivatives. For glucosyl-3-keto-ceramide and glucosyl-dihydro-ceramide (D-erythro), higher K,,, values were found than for glucosylceramide. Sphingosine, glucosylsphingosine and glucosyl-N-acetyl-sphingosine were the most effective inhibitors of the hydrolysis of glucosylceramide. D-erythro-Ceramide and D-galactosyl-N-acetyl-Derythro-sphingosine inhibited the hydrolyis of amphiphilic glucosylceramide but not that of water-soluble 4-methyl-umbelliferyl-B-glucoside, suggesting a hydrophobic binding site of the enzyme for the aglycon moiety of its membrane-bound substrate.Dilution experiments suggested that at least a fraction of the enzyme associates with the liposomes and degrades the lipid substrate even in the absence of activator proteins.Acidic phospholipids incorporated into liposomes caused a powerful stimulation (30 -40-fold) of the glucosylceramide P-glucosidase, whereas acidic sphingolipids (sulphatide, gangliosides GMl and GD incorporated into liposomes stimulated this enzyme only moderately (3 -10-fold).Lysosomal glucosylceramide 0-glucosidase (glucosylceramidase, EC 3.2.1.45) catalyzes the hydrolysis of glucosylceramide to ceramide and glucose. The activity of this 'acid' b-glucosidase is deficient in patients with the various forms of Gaucher's disease [l, 21. In order to gain more insight into the molecular basis and into the pathobiochemistry of this inherited disease, the cDNA for the human enzyme has been cloned [3], its post-translational modifications have been studied [4,5] and its activity with the synthetic substrate 4-methylumbelliferyl-P-D-glucopyranoside (4-MuGlc) [6, 7] has been investigated as well as its specificity for glucosylceramide in the presence of detergents [8 -111. A fluorescent derivative of glycosylceramide has been used for assaying glucosylceramidase activity of cultured cells in the presence and in the absence of detergents [12].Kinetic studies with glucosylceramide as substrate were performed in the presence of detergents in order to solubilize In the present work it was tested whether the hghly purified human glucosylceramide B-glucosidase, which is water-soluble, also recognizes glucosylceramide as substrate directly when the latter is inserted into lipid bilayers, in the absence of dispersing detergents or acti...

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“…Glucocerebrosidase was identified with acid f-glucosidase activity measured with the synthetic substrate 4-methylumbelliferyl ,-glucoside (Ho, 1973). Both enzyme activities depended on the association of two inactive macromolecular components, factors P and C (Ho & O'Brien, 1971;Ho et al, 1973a). Factor P was a soluble heat-stable glycoprotein fraction isolated from the spleen of a patient with adult Gaucher's disease.…”

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confidence: 99%

Glucocerebrosidase: reconstitution from macromolecular components depends on acidic phospholipids (Short Communication)

Light

1973

Biochemical Journal

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Glucocerebrosidase: reconstitution of activity from macromolecular components (Short Communication)

Abstract: Glucocerebrosidase activity was reconstituted in vitro from a soluble glycoprotein factor (P) and a particle-bound factor (C). The physiological significance of the system is discussed.

Cited by 82 publications

References 11 publications

Multiple Forms of Glycosidases in the Normal and Pathological States

Multiple Forms of Glycosidases in the Normal and Pathological States

Specificity of human glucosylceramide β‐glucosidase towards synthetic glucosylsphingolipids inserted into liposomes

Glucocerebrosidase: reconstitution from macromolecular components depends on acidic phospholipids (Short Communication)

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