title: In situ assessment of beta-hexosaminidase activity.
abstract: We have adapted two methods to evaluate the beta-hexosaminidase (HEX) enzymatic activity in cultured cells, based on the use of (i) the fluorogenic substrate 4-methylumbelliferyl-6-sulfo-2-acetamido-2- deoxy-beta-D-glucopyranoside (MU-GlcNAc-6-SO4) and (ii) the naphthol AS-BI-N-acetyl-beta-D-glucosaminide and hexazotized pararosaniline. We demonstrate that both methods could be used for the HEX isoenzymes by comparing wild-type and mutant human fibroblast cell lines, deficient for either an alpha or beta subunit from Tay-Sachs and Sandhoff patients. This in situ cytochemical assessment of HEX activity offers a rapid evaluation to study the expression of this enzyme in a heterogeneous cell population such as in gene transfer experiments.
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D002478:Cells, Cultured; D005347:Fibroblasts / Q000201:enzymology; D005965:Glucuronates / Q000378:metabolism; D006651:Histocytochemistry; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D009284:Naphthols; D012497:Sandhoff Disease / Q000201:enzymology*; D013050:Spectrometry, Fluorescence; D013661:Tay-Sachs Disease / Q000201:enzymology; D001619:beta-N-Acetylhexosaminidases / Q000235:genetics / Q000378:metabolism*
chemical_list: D005965:Glucuronates; D007527:Isoenzymes; D009284:Naphthols; C097027:naphthol AS-BI-N-acetyl-beta-glucuronide; D006923:Hymecromone; C043647:4-methylumbelliferyl-6-sulfo-2-acetamido-2-deoxy-beta-glucopyranoside; D001619:beta-N-Acetylhexosaminidases; C008584:naphthol AS; D000117:Acetylglucosamine