Activators of spleen glucocerebrosidase from controls and patients with various forms of Gaucher's disease

A. Basu, R. H. Glew, L. B. Daniels, L. S. Clark

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Glucocerebrosidase from normal human spleen, and spleen from cases of neurologic (types 2 and 3) and nonneurologic (type 1) Gaucher's disease, was delipidated and inactivated by extraction from membranes with sodium cholate and ice-cold 1-butanol. Control glucocerebrosidase was stimulated markedly by large quantities (20-30 μg/assay) of phosphatidylserine (PS), or by a combination of smaller amounts (1-2 μg) of PS and 3 μg of a heat-stable factor (HSF) derived from the spleen of a patient with Gaucher's disease. The residual glucocerebrosidase from a nonneurologic case, but not a neurologic case, was also responsive to PS and HSF. The combination of HSF and PS decreased the K(m) of the normal enzyme for 4-methylum-belliferyl-β-D-glucopyranoside from 8.0 to 1.6 mM. These effectors also increased the reactivity of glucocerebrosidase to the inhibitor conduritol B expoxide; HSF alone had no effect (t 1/2 = 19 ± 0.5 min) whereas the maximum rate of inactivation (t 1/2 = 4.0 min) by conduritol B epoxide was achieved in the presence of a mixture of PS (1 μg) and HSF (3 μg). Phosphatidylglycerol (PG) and phosphatidic acid, also acidic phospholipids, were effective activators of glucocerebrosidase. Varying the fatty acid composition of PG had little effect on its ability to stimulate glucocerebrosidase activity. However, in the case of phosphatidylcholine (PC), a weaker activator than PG or PS, fatty acid composition had a significant impact on the ability of this neutral lipid to activate glucocerebrosidase; dilinoleoyl-PC and dicaproyl-PC were moderately effective activators, but distearoyl-PC and dioleoyl-PC were almost totally inactive. The mono-, and di-, and trisialogangliosides (G(M1), G(D1), and G(T1) were less than half as effective as PS as activators of glucocerebrosidase. These results indicate that acidic phospholipids and the heat-stable factor may both play a role in explaining the genetic heterogeneity of Gaucher's disease.

Original languageEnglish
Pages (from-to)1714-1719
Number of pages6
JournalJournal of Biological Chemistry
Volume259
Issue number3
StatePublished - 1 Jan 1984

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Glucosylceramidase
Gaucher Disease
Phosphatidylserines
Spleen
Hot Temperature
Phosphatidylglycerols
Phosphatidylcholines
1,2-linoleoylphosphatidylcholine
Nervous System
Phospholipids
Fatty Acids
Sodium Cholate
Phosphatidic Acids
1-Butanol
Genetic Heterogeneity
Ice
Chemical analysis
Assays
Membranes
Lipids

Cite this

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title = "Activators of spleen glucocerebrosidase from controls and patients with various forms of Gaucher's disease",
abstract = "Glucocerebrosidase from normal human spleen, and spleen from cases of neurologic (types 2 and 3) and nonneurologic (type 1) Gaucher's disease, was delipidated and inactivated by extraction from membranes with sodium cholate and ice-cold 1-butanol. Control glucocerebrosidase was stimulated markedly by large quantities (20-30 μg/assay) of phosphatidylserine (PS), or by a combination of smaller amounts (1-2 μg) of PS and 3 μg of a heat-stable factor (HSF) derived from the spleen of a patient with Gaucher's disease. The residual glucocerebrosidase from a nonneurologic case, but not a neurologic case, was also responsive to PS and HSF. The combination of HSF and PS decreased the K(m) of the normal enzyme for 4-methylum-belliferyl-β-D-glucopyranoside from 8.0 to 1.6 mM. These effectors also increased the reactivity of glucocerebrosidase to the inhibitor conduritol B expoxide; HSF alone had no effect (t 1/2 = 19 ± 0.5 min) whereas the maximum rate of inactivation (t 1/2 = 4.0 min) by conduritol B epoxide was achieved in the presence of a mixture of PS (1 μg) and HSF (3 μg). Phosphatidylglycerol (PG) and phosphatidic acid, also acidic phospholipids, were effective activators of glucocerebrosidase. Varying the fatty acid composition of PG had little effect on its ability to stimulate glucocerebrosidase activity. However, in the case of phosphatidylcholine (PC), a weaker activator than PG or PS, fatty acid composition had a significant impact on the ability of this neutral lipid to activate glucocerebrosidase; dilinoleoyl-PC and dicaproyl-PC were moderately effective activators, but distearoyl-PC and dioleoyl-PC were almost totally inactive. The mono-, and di-, and trisialogangliosides (G(M1), G(D1), and G(T1) were less than half as effective as PS as activators of glucocerebrosidase. These results indicate that acidic phospholipids and the heat-stable factor may both play a role in explaining the genetic heterogeneity of Gaucher's disease.",
author = "A. Basu and Glew, {R. H.} and Daniels, {L. B.} and Clark, {L. S.}",
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Activators of spleen glucocerebrosidase from controls and patients with various forms of Gaucher's disease. / Basu, A.; Glew, R. H.; Daniels, L. B.; Clark, L. S.

In: Journal of Biological Chemistry, Vol. 259, No. 3, 01.01.1984, p. 1714-1719.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Activators of spleen glucocerebrosidase from controls and patients with various forms of Gaucher's disease

AU - Basu, A.

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N2 - Glucocerebrosidase from normal human spleen, and spleen from cases of neurologic (types 2 and 3) and nonneurologic (type 1) Gaucher's disease, was delipidated and inactivated by extraction from membranes with sodium cholate and ice-cold 1-butanol. Control glucocerebrosidase was stimulated markedly by large quantities (20-30 μg/assay) of phosphatidylserine (PS), or by a combination of smaller amounts (1-2 μg) of PS and 3 μg of a heat-stable factor (HSF) derived from the spleen of a patient with Gaucher's disease. The residual glucocerebrosidase from a nonneurologic case, but not a neurologic case, was also responsive to PS and HSF. The combination of HSF and PS decreased the K(m) of the normal enzyme for 4-methylum-belliferyl-β-D-glucopyranoside from 8.0 to 1.6 mM. These effectors also increased the reactivity of glucocerebrosidase to the inhibitor conduritol B expoxide; HSF alone had no effect (t 1/2 = 19 ± 0.5 min) whereas the maximum rate of inactivation (t 1/2 = 4.0 min) by conduritol B epoxide was achieved in the presence of a mixture of PS (1 μg) and HSF (3 μg). Phosphatidylglycerol (PG) and phosphatidic acid, also acidic phospholipids, were effective activators of glucocerebrosidase. Varying the fatty acid composition of PG had little effect on its ability to stimulate glucocerebrosidase activity. However, in the case of phosphatidylcholine (PC), a weaker activator than PG or PS, fatty acid composition had a significant impact on the ability of this neutral lipid to activate glucocerebrosidase; dilinoleoyl-PC and dicaproyl-PC were moderately effective activators, but distearoyl-PC and dioleoyl-PC were almost totally inactive. The mono-, and di-, and trisialogangliosides (G(M1), G(D1), and G(T1) were less than half as effective as PS as activators of glucocerebrosidase. These results indicate that acidic phospholipids and the heat-stable factor may both play a role in explaining the genetic heterogeneity of Gaucher's disease.

AB - Glucocerebrosidase from normal human spleen, and spleen from cases of neurologic (types 2 and 3) and nonneurologic (type 1) Gaucher's disease, was delipidated and inactivated by extraction from membranes with sodium cholate and ice-cold 1-butanol. Control glucocerebrosidase was stimulated markedly by large quantities (20-30 μg/assay) of phosphatidylserine (PS), or by a combination of smaller amounts (1-2 μg) of PS and 3 μg of a heat-stable factor (HSF) derived from the spleen of a patient with Gaucher's disease. The residual glucocerebrosidase from a nonneurologic case, but not a neurologic case, was also responsive to PS and HSF. The combination of HSF and PS decreased the K(m) of the normal enzyme for 4-methylum-belliferyl-β-D-glucopyranoside from 8.0 to 1.6 mM. These effectors also increased the reactivity of glucocerebrosidase to the inhibitor conduritol B expoxide; HSF alone had no effect (t 1/2 = 19 ± 0.5 min) whereas the maximum rate of inactivation (t 1/2 = 4.0 min) by conduritol B epoxide was achieved in the presence of a mixture of PS (1 μg) and HSF (3 μg). Phosphatidylglycerol (PG) and phosphatidic acid, also acidic phospholipids, were effective activators of glucocerebrosidase. Varying the fatty acid composition of PG had little effect on its ability to stimulate glucocerebrosidase activity. However, in the case of phosphatidylcholine (PC), a weaker activator than PG or PS, fatty acid composition had a significant impact on the ability of this neutral lipid to activate glucocerebrosidase; dilinoleoyl-PC and dicaproyl-PC were moderately effective activators, but distearoyl-PC and dioleoyl-PC were almost totally inactive. The mono-, and di-, and trisialogangliosides (G(M1), G(D1), and G(T1) were less than half as effective as PS as activators of glucocerebrosidase. These results indicate that acidic phospholipids and the heat-stable factor may both play a role in explaining the genetic heterogeneity of Gaucher's disease.

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