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American Journal of Human Genetics Jul 1980I-cell fibroblasts with a multiple intracellular lysosomal enzyme deficiency were hybridized with cells from patients with different types of single lysosomal enzyme...
I-cell fibroblasts with a multiple intracellular lysosomal enzyme deficiency were hybridized with cells from patients with different types of single lysosomal enzyme defects. Fusion with G(M2) gangliosidosis, type 2, (Sandhoff disease) fibroblasts resulted in a restoration of the hexosaminidase activity, in a normalization of the electrophoretic mobility of the isoenzymes, and in a decreased activity in the medium. Fusion of I-cells with fibroblasts from G(M1) gangliosidosis, type 1, led to enhancement of beta-galactosidase (beta-gal) activity. This complementation must be the result of the presence of normal polypeptide chains in I-cells, whereas the other cell types provide a factor that causes the intracellular retention of the enzymes. Restoration of beta-gal was also observed in heterokaryons after fusion of I-cells with beta-galactosidase/neuraminidase-deficient (beta-gal(-)/neur(-)) variants, indicating that the neuraminidase(s) and the posttranslational modification of beta-gal are affected in a different way in I-cell disease and in beta-gal(-)/neur(-) variants. Fusion of I-cells with mannosidosis fibroblasts resulted in a restoration of the acidic form of alpha-mannosidase and in a decrease of the extracellular activity of both this enzyme and the hexosaminidase enzyme, indicating that fusion of I-cells with different types of fibroblasts with a single lysosomal enzyme deficiency not only leads to complementation for one particular enzyme but also to a correction of the basic defect in I-cells.
Topics: Cell Fusion; Cells, Cultured; Fibroblasts; Gangliosidoses; Genetic Complementation Test; Hexosaminidases; Humans; Hybrid Cells; Lysosomes; Mannosidases; Mucolipidoses; Sandhoff Disease; beta-Galactosidase
PubMed: 6772024
DOI: No ID Found -
The Journal of Clinical Investigation Sep 1975Patients with mannosidosis, an inherited deficiency of lysosomal alpha-mannosidase, accumulate large amounts of mannose-rich oligosaccharides (the "core" of the...
Patients with mannosidosis, an inherited deficiency of lysosomal alpha-mannosidase, accumulate large amounts of mannose-rich oligosaccharides (the "core" of the carbohydrate units of many glocoproteins) in brain and liver and excrete these partial degradation products in their urine. A profound alpha-mannosidase deficiency was demonstrated in fibroblasts cultured from a skin biopsy obtained from a child with mannosidosis. Further, abnormal glycopeptides rich in mannose and similar to oligosaccharides found in the patient's urine were isolated from fibroblast extracts by a variety of chromatographic procedures and by virtue of their binding to a concanavalin A-Sepharose 4B affinity column. This storage material contained mannose, N-acetylglucosamine, and asparagine in the ratio 3 : 1 : 1 together with a few toher amino acids and had a molecular weight of approximately 1,100. There was no evidence for excretion of storage material by mannosidosis fibroblasts or for any abnormality in cell surface glycoprotein composition. The glycopeptide nature of the storage material isolated from cultured skin fibroblasts may be attributed to the low level of N-aspartyl-beta-glucosamindase (EC 3.5.1.-) activity in these cells.
Topics: Asparaginase; Aspartylglucosylaminase; Cells, Cultured; Child, Preschool; Disaccharidases; Extracellular Space; Fibroblasts; Glucosamine; Glycopeptides; Glycoproteins; Glycosaminoglycans; Glycosphingolipids; Humans; Male; Mannose; Mannosidases; Mucopolysaccharidoses; Skin
PubMed: 125765
DOI: 10.1172/JCI108142 -
The Biochemical Journal Feb 1986Two homologous series of urinary oligosaccharides were identified by h.p.l.c. and fast-atom-bombardment mass spectrometry in feline alpha-mannosidosis. The predominant...
Two homologous series of urinary oligosaccharides were identified by h.p.l.c. and fast-atom-bombardment mass spectrometry in feline alpha-mannosidosis. The predominant series has the composition Man2-8GlcNAc2 and a minor series the composition Man2-9GlcNAc. The structure of the most abundant oligosaccharide, which accounts for over 80% of the urinary oligosaccharide, was shown to be alpha-D-Manp(1----3)[alpha-D-Manp-(1----6)]beta-D-Manp -(1----4)-beta-D-GlcpNA c-(1----4)-D-GlcNAc by gas chromatography and mass spectrometry. Such a structure is consistent with the incomplete catabolism of complex N-linked glycans due to a deficiency of alpha-D-mannosidase in tissue lacking an endohexosaminidase activity.
Topics: Animals; Cats; Chromatography, Gas; Chromatography, Gel; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Methylation; Oligosaccharides; alpha-Mannosidosis
PubMed: 3707532
DOI: 10.1042/bj2330899 -
FEBS Letters Oct 1983Two major oligosaccharides were isolated by preparative HPLC from the urine of a locoweed-fed sheep. Analysis by gas-liquid chromatography and mass-spectrometry...
Two major oligosaccharides were isolated by preparative HPLC from the urine of a locoweed-fed sheep. Analysis by gas-liquid chromatography and mass-spectrometry indicated compositions of (Man)4(GlcNAc)2 and (Man)5(GlcNAc)2, respectively. Structures were determined by digestion with alpha-D-mannosidase and endo-beta-N-acetylglucosaminidases D and H, and comparison of the products by HPLC with synthetic standards, and oligosaccharides isolated from human mannosidosis urine. Incubation with an exo-beta-N-acetylglucosaminidase was without effect.
Topics: Acetylglucosamine; Animals; Carbohydrate Conformation; Carbohydrate Sequence; Chromatography, High Pressure Liquid; Female; Gas Chromatography-Mass Spectrometry; Mannose; Oligosaccharides; Plant Poisoning; Sheep; Sheep Diseases
PubMed: 6628681
DOI: 10.1016/0014-5793(83)81173-9 -
Annals of Saudi Medicine 2004
Topics: Anticonvulsants; Child, Preschool; Consanguinity; Epilepsy; Female; Humans; Intellectual Disability; Speech Disorders; beta-Mannosidosis
PubMed: 15573858
DOI: 10.5144/0256-4947.2004.393 -
European Journal of Biochemistry Nov 1975Abnormally high amounts of low molecular weight mannose-rich carbohydrate material were found in the urine of an Angus calf with mannosidosis. At least five...
Abnormally high amounts of low molecular weight mannose-rich carbohydrate material were found in the urine of an Angus calf with mannosidosis. At least five oligosaccharide fractions were detected by paper chromatography. The most abundant compound was purified by gel chromatography, zone electrophoresis, and two consecutive preparative paper chromatographic steps. The yield was 10 mg/liter of urine. From structural studies including nuclear magnetic resonance spectroscopy, optical rotation, sugar analysis, methylation analysis, and partial enzymatic degradation the following structure was deduced: alpha-D-Manp-(1 leads to 6)-beta-D-Manp-(1 leads to 4)-beta-D-GlcNAcp-(1 leads to 4)-beta-D-GlcNAcp-(1 leads to 4)-D-GlcNAc. This oligosaccharide is distinct from all the oligosaccharides previously described which are excreted by patients with mannosidosis.
Topics: Acetylglucosamine; Animals; Carbohydrate Metabolism, Inborn Errors; Cattle; Cattle Diseases; Magnetic Resonance Spectroscopy; Mannose; Mannosides; Oligosaccharides
PubMed: 1204626
DOI: 10.1111/j.1432-1033.1975.tb02488.x