GM1-ganglioside β-galactosidase in leukocytes and cultured fibroblasts

Raghavan, Subharekha; Gajewski, Anna; Kolodny, Edwin H.

doi:10.1016/0009-8981(77)90412-0

Cited by 13 publications

(4 citation statements)

References 21 publications

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“…The activity of G M1 (II 3 -␣ -Neu5Ac-Gg 4 Cer) ␤ -galactosidase (EC [enzyme commission number] 3.2.1.23) was measured by using 4-MUF-␤ -D-galactopyranoside as a substrate (23); the activity of G M2 (II 3 -␣ -Neu5Ac-Gg 3 Cer) ␤ -hexosaminidase (EC 3.2.1.52) was measured by using 4-MUF-glucosaminide as a substrate (24), or 4-MUF-N-acetylglucosamine-6-sulfate for izoenzyme ␤ -hexosaminidase A activity (25). The activity of arylsulfatase A (ASA; EC 3.1.6.1) involved in sulfatide degradation was measured by spectrophotometry ( ϭ 515 nm), using chromogenic substrate p -nitrocatechol sulfate (26).…”

Section: Determination Of Lysosomal Enzyme Activities In Leukocytesmentioning

confidence: 99%

Leukocyte Lysosomal Enzymes in Alzheimer's Disease and Down's Syndrome

Kalanj-Bognar¹,

Rundek²,

Furač³

et al. 2002

The Journals of Gerontology Series A: Biological Sciences and M

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Previous studies suggested the possibility of accelerated lysosomal degradation of brain gangliosides in Alzheimer's disease (AD). As AD pathology affects both neural and nonneural tissues, the aim of this study was to determine possible changes of glycosphingolipid metabolism in available peripheral cells in AD and Down's syndrome (DS). The activities of several lysosomal enzymes involved in catabolism of gangliosides and sulfatides were measured in leukocytes from subjects with dementia of the Alzheimer type, DS, and age-matched controls, by fluorimetry and spectrophotometry using specific substrates. The results showed a statistically significant increase of ␤ -galactosidase activity in both dementia of the Alzheimer type and DS leukocytes when compared with age-matched controls ( p Ͻ .01 and p Ͻ .05, respectively; Student's t test). Not significantly increased activities of ␤ -galactosidase, ␤ -hexosaminidase, ␤ -hexosaminidase A, and slightly decreased activity of arylsulfatase A were observed in control leukocytes with aging. Our results indicate that a metabolic dysfunction and the acceleration of at least some lysosomal catabolic pathways are present in AD and DS nonneural cells. ć LZHEIMER'S disease (AD) is a neurodegenerative disorder of unknown etiology. The hypothesis of disturbed glycosphingolipid (GSL) metabolism in AD is based on several facts. First, previous studies showed changes in content and composition of brain gangliosides in Alzheimer's disease (1), and they suggested the possibility of accelerated lysosomal degradation of GSLs in AD brains (2). Second, numerous studies have shown that pathologic processes and biochemical disturbances in AD affect not only neural but also nonneural tissues (3-10). Third, the majority of biochemical disorders in neural and nonneural tissue result in seriously damaged structure, function, and fluidity of cell membranes (11-15). Finally, gangliosidesGSLs containing sialic acids-are incorporated in all animal cell membranes, with the highest content, variety, and specific regional distribution found in the human brain tissue. As important membrane constituents, GSLs confer to its structural rigidity and are also (themselves or as intermediate metabolites) key players in processes such as cell growth, proliferation, differentiation, cell recognition, and apoptosis (16-20).Down's syndrome (DS) is used as a comparative model for the study of AD: the same clinical symptoms of dementia and the neuropathological hallmarks characteristic of AD are present in older DS individuals (21). Also, similar biochemical disorders have been observed in both AD and DS nonneural tissues (7).The aim of this study was to determine possible changes of GSL metabolism in available peripheral cells derived from subjects with a clinical diagnosis of dementia of the Alzheimer type (DAT) and DS. M ETHODS SubjectsBlood samples were collected at the Department for Neurology, University Hospital "Sestre milosrdnice" in Zagreb, from 19 patients (7 men and 12 women, age range 47-86 years; Tab...

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Section: Determination Of Lysosomal Enzyme Activities In Leukocytesmentioning

confidence: 99%

Leukocyte Lysosomal Enzymes in Alzheimer's Disease and Down's Syndrome

Kalanj-Bognar¹,

Rundek²,

Furač³

et al. 2002

The Journals of Gerontology Series A: Biological Sciences and M

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“…Although a very low concentration of heparin may be present during assays measuring plasma enzyme activities, only traces of heparin (if any) are expected after the leukocyte preparation procedure. However, there are many biochemical genetic laboratories which use heparin as anticoagulant (3,4,(19)(20)(21), and some reference centers recommend the collection of blood with heparin concentrations as high as 10% (Shin Y, personal communication). In the present study, higher lysosomal enzyme activities were observed when blood was collected with 10% heparin.…”

Section: Discussionmentioning

confidence: 99%

Effect of collection, transport, processing and storage of blood specimens on the activity of lysosomal enzymes in plasma and leukocytes

Burin

Dutra-Filho

Brum³

et al. 2000

Braz J Med Biol Res

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This study was designed to evaluate the effect of different conditions of collection, transport and storage on the quality of blood samples from normal individuals in terms of the activity of the enzymes ß-glucuronidase, total hexosaminidase, hexosaminidase A, arylsulfatase A and ß-galactosidase. The enzyme activities were not affected by the different materials used for collection (plastic syringes or vacuum glass tubes). In the evaluation of different heparin concentrations (10% heparin, 5% heparin, and heparinized syringe) in the syringes, it was observed that higher doses resulted in an increase of at least 1-fold in the activities of ß-galactosidase, total hexosaminidase and hexosaminidase A in leukocytes, and ß-glucuronidase in plasma. When the effects of time and means of transportation were studied, samples that had been kept at room temperature showed higher deterioration with time (72 and 96 h) before processing, and in this case it was impossible to isolate leukocytes from most samples. Comparison of heparin and acid citrate-dextrose (ACD) as anticoagulants revealed that ß-glucuronidase and hexosaminidase activities in plasma reached levels near the lower normal limits when ACD was used. In conclusion, we observed that heparin should be used as the preferable anticoagulant when measuring these lysosomal enzyme activities, and we recommend that, when transport time is more than 24 h, samples should be shipped by air in a styrofoam box containing wet ice.

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“…GH, ganglioside p-galactosidase activity was assayed by the method of Raghavan et a1 [25] and 4-methylumbelliferyl P-galactosidase by previously reported methods…”

Section: Methodsmentioning

confidence: 99%

“…The [:'HI-labeled gangliosides were fractionated by column chromatography [34] and the [?HI GM, ganglioside fractions further purified by preparative thin-layer chromatography using the tetrahydrofurad water/potassium chloride solvent system described by Eberlein and Gucken [5]. The specific activity of ["HI G M , ganglioside was adjusted to 2,500 cpm per nanomole with unlabeled GMi ganglioside.GH, ganglioside p-galactosidase activity was assayed by the method of Raghavan et a1 [25] and 4-methylumbelliferyl P-galactosidase by previously reported methods Proteins were determined by the method of Lowry et a1 [14]. Urinary oligosaccharides were assayed qualitatively…”

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confidence: 99%

G_M1 Gangliosidosis: Phenotypic variation in a single family

Farrell

Ochs

1981

Annals of Neurology

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Infantile, juvenile, and adult forms of GM1 gangliosidosis have been well characterized. Certain genetic and biochemical studies have suggested that the phenotypic variation found in GM1 gangliosidosis results from different allelic mutations affecting the GM1 ganglioside beta-galactosidase locus and that different combinations of these mutations accounts for the clinical heterogeneity of this illness. A family in which both the infantile and juvenile forms of GM1 gangliosidosis occurred, the children sharing a common mutation of their acid beta-galactosidase activity, supports the allelic nature of these different clinical forms of the disease. From the observations made in this unique family, additional phenotypes of GM1 gangliosidosis might be anticipated.

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GM1-ganglioside β-galactosidase in leukocytes and cultured fibroblasts

Cited by 13 publications

References 21 publications

Leukocyte Lysosomal Enzymes in Alzheimer's Disease and Down's Syndrome

Leukocyte Lysosomal Enzymes in Alzheimer's Disease and Down's Syndrome

Effect of collection, transport, processing and storage of blood specimens on the activity of lysosomal enzymes in plasma and leukocytes

G_M1 Gangliosidosis: Phenotypic variation in a single family

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GM1-ganglioside β-galactosidase in leukocytes and cultured fibroblasts

Cited by 13 publications

References 21 publications

Leukocyte Lysosomal Enzymes in Alzheimer's Disease and Down's Syndrome

Leukocyte Lysosomal Enzymes in Alzheimer's Disease and Down's Syndrome

Effect of collection, transport, processing and storage of blood specimens on the activity of lysosomal enzymes in plasma and leukocytes

GM1 Gangliosidosis: Phenotypic variation in a single family

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G_M1 Gangliosidosis: Phenotypic variation in a single family