title: Alpha-L-fucosidase in cultured bone marrow fibroblasts from fucosidosis patients.
abstract: Bone marrow fibroblasts were cultured from two patients with fucosidosis type 2, six control subjects, and three patients with other lysosomal disorders. Optimal conditions for measuring alpha-L-fucosidase activity in lysates of these cells with the fluorogenic substrate 4-methylumbelliferyl-alpha-L-fucoside were established. The pH profile of normal bone marrow fibroblasts showed three peaks and a shoulder of enzymatic activity, with maximum activity at pH 4.75. In cells derived from fucosidosis patients two peaks of apparent alpha-L-fucosidase activity were obtained; the pH optimum was 4.5. alpha-L-Fucosidase activity (mean +/- SD) in the fucosidosis and control bone marrow fibroblasts was 2.5 and 312.4 +/- 10.9 nmoles 4-methylumbelliferone per milligram protein per hour, respectively. A reduction in the apparent specific enzymatic activity in the fucosidosis cells was observed by using increasing concentrations of cellular protein in the assay system. Mixing experiments between normal and fucosidosis cells gave the expected activities. These findings indicate that cultured bone marrow fibroblasts can be used for the diagnosis and study of fucosidosis.
mesh_terms: D001853:Bone Marrow / Q000473:pathology; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D005347:Fibroblasts / Q000201:enzymology; D005776:Gaucher Disease / Q000201:enzymology; D006801:Humans; D007966:Leukodystrophy, Metachromatic / Q000201:enzymology; D008361:Mannosidases / Q000032:analysis; D009083:Mucopolysaccharidoses / Q000201:enzymology; D005644:alpha-L-Fucosidase; D043323:alpha-Mannosidase; D001616:beta-Galactosidase / Q000032:analysis
chemical_list: D008361:Mannosidases; D001616:beta-Galactosidase; D043323:alpha-Mannosidase; D005644:alpha-L-Fucosidase