title: [Use of fluorescence-labelled cerebroside as a substrate for galactosylceramidase of human skin fibroblasts].
abstract: Possible use of the fluorescently labelled cerebroside, 1-O-(beta-D-galactosyl)-2-N-[6-(2-antroyl)hexanoyl]-4-sphingeni n (Gal-A-sphingenin) as a substrate for galactosylceramidase (GC) from human skin fibroblasts was investigated. Studies involving TLC and fluorimetric methods revealed enzymatic splitting of Gal-A-sphingenin whose degree correlated with the amount of the enzyme and incubation time. Some kinetic parameters of GC were determined, using Gal-A-sphingenin as substrate. It was shown that the values of specific activity of GC (1.6 nmol/mg protein/hr) and Km (0.025 mM) for Gal-A-sphingenin agree well with the corresponding values obtained with the use of the galactocerebroside as a natural substrate. In experiments with mixtures of Gal-A-sphingening and galactocerebroside used as substrates in different molar ratios, it was shown that the enzyme splits each of the substrates with equal velocity. The results of experiments with the enzyme samples from skin fibroblasts of healthy individuals and of patients with GM1-gangliosidosis (GM1-beta-D-galactosidase deficiency) suggest that Gal-A-sphingenin is a specific substrate for GC.
mesh_terms: D004789:Enzyme Activation; D005347:Fibroblasts / Q000201:enzymology; D005456:Fluorescent Dyes; D005696:Galactosidases / Q000378:metabolism*; D005698:Galactosylceramidase / Q000378:metabolism*; D006801:Humans; D011609:Psychosine / Q000031:analogs & derivatives* / Q000378:metabolism; D012867:Skin / Q000201:enzymology*; D013110:Sphingosine / Q000031:analogs & derivatives*; D013379:Substrate Specificity; D013661:Tay-Sachs Disease / Q000201:enzymology
chemical_list: D005456:Fluorescent Dyes; D011609:Psychosine; C044168:1-O-galactosyl-2-N-(6-(2-anthroyl)hexanoyl)-4-sphingenine; D005696:Galactosidases; D005698:Galactosylceramidase; D013110:Sphingosine
vernacular_title: Ispol'zovanie fluorestsentno mechennogo tserebrozida v kachestve substrata dlia galaktoziltseramidazy fibroblastov kozhi cheloveka.