Ganglioside GM2 N-acetyl-beta-D-galactosaminidase activity in cultured fibroblasts of late-infantile and adult GM2 gangliosidosis patients and of healthy probands with low hexosaminidase level.
A sensitive assay was developed to assess the ability of extracts from cultured fibroblasts to catabolize ganglioside GM2, in the presence of the natural activator protein but without detergents. This method, which permitted the reliable determination of residual activities as low as 0.1% of normal controls, was then used to measure ganglioside GM2 hydrolase activities in fibroblasts from several hexosaminidase variants. The residual activities thus determined correlated well with the clinical status of the respective proband: infantile Tay-Sachs (0.1% of normal controls), late-infantile (0.5%), and adult GM2 gangliosidoses (2%-4%) and healthy probands with "low hexosaminidase" (11% and 20%). In contrast, beta-hexosaminidase A levels as measured with the synthetic substrate 4-MU-GlcNAc could not be relied on for diagnostic purposes (the late-infantile patient studied retained 80% of the activity of controls).
D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D049289:G(M2) Activator Protein; D005678:G(M2) Ganglioside; D014644:Genetic Variation; D006023:Glycoproteins; D006596:Hexosaminidases / Q000172:deficiency* / Q000378:metabolism*; D006801:Humans; D007525:Isoelectric Focusing; D008297:Male; D011488:Protein Deficiency; D049231:Saposins; D049128:Sphingolipid Activator Proteins; D013661:Tay-Sachs Disease / Q000201:enzymology*
D049289:G(M2) Activator Protein; D006023:Glycoproteins; C483345:PSAP protein, human; D049231:Saposins; D049128:Sphingolipid Activator Proteins; D005678:G(M2) Ganglioside; C028960:GM2-N-acetyl-beta-D-galactosaminidase; D006596:Hexosaminidases