title: Molecular forms of beta-N-acetylhexosaminidase in Epstein-Barr virus-transformed lymphoid cell lines from normal subjects and patients with Tay-Sachs disease.
abstract: In whole leukocytes and in lymphocytes from normal subjects, the percentage activity of heat-stable beta-N-acetylhexosaminidase (30 +/- 5% and 45 +/- 5%, respectively) was higher than in the transformed lymphoid cell line (19 +/- 3%). In Tay-Sachs transformed cells as well as non-transformed beta-N-acetylhexosaminidase was almost completely heat-stable (95 - 98%). In the transformed cells from normal subjects, the beta-N-acetylhexosaminidase B (Hex B) activity (5% of total) was significantly lower than in blood lymphocytes (average 25 - 30% of total activity), whereas Hex A and Hex I were similar in the either cell type. Blood lymphocytes and lymphoid cell lines established from a Tay-Sachs patient lacked heat-labile Hex A and expressed high heat-stable Hex I and Hex B activities (3-6-fold). After neuraminidase treatment, Hex A peak sharpened while Hex I peaks switched to higher pI than normal Hex I, in the region of Hex B. PreHex A/S pI was not affected. Hydrolytic properties using the both substrates (4-methylumbelliferyl-2-acetamido-2-deoxy-beta-D-glucopyranoside and 4-methylumbelliferyl-2-acetamido-2-deoxy-beta-D-galactopyranoside) of each molecular form were similar in transformed and non-transformed cells. Data derived from the use of a mixture of substrates were consistent with the model which proposes a common active site for either substrate in the case of preHex A, Hex B and Hex I, but not for Hex A. Thus Epstein-Barr virus-transformed lymphoid cell lines represent an accurate model system for studies on Tay-Sachs disease.
mesh_terms: D002460:Cell Line; D002472:Cell Transformation, Viral; D004854:Herpesvirus 4, Human; D054818:Hexosaminidase A; D054819:Hexosaminidase B; D006596:Hexosaminidases / Q000378:metabolism*; D006801:Humans; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000378:metabolism; D007962:Leukocytes / Q000201:enzymology; D008214:Lymphocytes / Q000201:enzymology*; D009439:Neuraminidase; D013661:Tay-Sachs Disease / Q000201:enzymology*; D001619:beta-N-Acetylhexosaminidases
chemical_list: D007527:Isoenzymes; D006596:Hexosaminidases; D009439:Neuraminidase; D054818:Hexosaminidase A; D054819:Hexosaminidase B; D001619:beta-N-Acetylhexosaminidases