d'Azzo A, Proia R L, Kolodny E H, Kaback M M, Neufeld E F
J Biol Chem. 1984 Sep 10;259(17):11070-4.
We have previously described the kinetics of association of the alpha- and beta-subunits of beta-hexosaminidase A in intact cultured human fibroblasts, using biosynthetic labeling and immunoprecipitation with antisera that distinguish between monomeric and associated alpha-chains (Proia, R. L., d'Azzo, A., and Neufeld, E. F. (1984) J. Biol. Chem. 259, 3350-3354). We now show lack of alpha-beta association in fibroblasts of several individuals deficient in beta-hexosaminidase A (5 patients with nonclassic forms of Tay-Sachs disease and 2 asymptomatic siblings). Defective association was accompanied by markedly reduced (less than one-tenth of normal) conversion of the alpha-chain precursor of Mr = 67,000 to the mature lysosomal form of Mr = 54,000. Analysis by hybridization with fibroblasts lacking the alpha- or beta-chain showed that the association defect resided in the alpha-chain. Most of the cell strains studied also had decreased synthesis of the alpha-chain, suggesting compound heterozygosity with the Ashkenazi Tay-Sachs (no synthesis) allele. An unusual feature of the association defect is the variability in the resulting clinical manifestations, even within families, implying that other factors determine the adequacy of the residual associated beta-hexosaminidase A in vivo.
我们先前已利用生物合成标记和能区分单体α链和结合型α链的抗血清进行免疫沉淀,描述了完整培养的人成纤维细胞中β-己糖胺酶A的α亚基和β亚基的结合动力学(Proia, R. L., d'Azzo, A., and Neufeld, E. F. (1984) J. Biol. Chem. 259, 3350 - 3354)。我们现在发现,在几名β-己糖胺酶A缺乏的个体的成纤维细胞中,α-β结合缺失(5例非典型泰-萨克斯病患者和2名无症状同胞)。结合缺陷伴随着Mr = 67,000的α链前体向Mr = 54,000的成熟溶酶体形式的显著减少(不到正常的十分之一)。与缺乏α链或β链的成纤维细胞杂交分析表明,结合缺陷存在于α链中。大多数研究的细胞系中α链的合成也减少,提示与阿什肯纳兹泰-萨克斯(无合成)等位基因存在复合杂合性。结合缺陷的一个不寻常特征是,即使在家族内部,所产生的临床表现也存在变异性,这意味着其他因素决定了体内残余结合型β-己糖胺酶A的充足性。