Conzelmann E, Kytzia H J, Navon R, Sandhoff K
Am J Hum Genet. 1983 Sep;35(5):900-13.
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).
我们开发了一种灵敏的检测方法,用于评估在存在天然激活蛋白但无去污剂的情况下,培养的成纤维细胞提取物分解神经节苷脂GM2的能力。该方法能够可靠地测定低至正常对照0.1%的残余活性,随后被用于测量几种己糖胺酶变体的成纤维细胞中神经节苷脂GM2水解酶的活性。由此测定的残余活性与各自先证者的临床状况密切相关:婴儿型泰-萨克斯病(为正常对照的0.1%)、晚婴儿型(0.5%)、成人GM2神经节苷脂沉积症(2%-4%)以及“低己糖胺酶”的健康先证者(11%和20%)。相比之下,用合成底物4-MU-GlcNAc测定的β-己糖胺酶A水平不能用于诊断目的(所研究的晚婴儿型患者保留了对照活性的80%)。