Aerts J M, Donker-Koopman W E, Brul S, Van Weely S, Sa Miranda M C, Barranger J A, Tager J M, Schram A W
E. C. Slater Institute for Biochemical Research, University of Amsterdam, The Netherlands.
Biochem J. 1990 Jul 1;269(1):93-100. doi: 10.1042/bj2690093.
In Gaucher disease (glucosylceramide lipidosis), deficiency of glucocerebrosidase causes pathological storage of glucosylceramide, particularly in the spleen. A comparative biochemical and immunological analysis has therefore been made of glucocerebrosidase in spleens from normal subjects (n = 4) and from Gaucher disease patients with non-neuronopathic (n = 5) and neuronopathic (n = 5) phenotypes. The spleens from all Gaucher disease patients showed markedly decreased glucocerebrosidase activity. Discrimination of different phenotypes of Gaucher disease was not possible on the basis of the level of residual enzyme activity, or by measurements, using the immunopurified enzyme, of kinetic constants, pI or molecular mass forms. A severe decrease was found in the specific activity of glucocerebrosidase purified to homogeneity from the spleen of a patient with the non-neuronopathic phenotype of Gaucher disease, as compared with that of the enzyme purified from the spleen of a normal subject. This finding was confirmed by an immunological method developed for accurate assessment of the relative enzyme activity per molecule of glucocerebrosidase protein. The method revealed that the residual enzyme in the spleens of all investigated patients with a non-neuronopathic course of Gaucher disease had a more than 7-fold decreased activity of glucocerebrosidase (measured in the presence of taurocholate) per molecule of enzyme, and that the concentration of glucocerebrosidase molecules in the spleens of these patients was near normal. Observations made with immunoblotting experiments were consistent with these findings. In contrast, in the spleens of patients with neuronopathic phenotypes of Gaucher disease, the concentration of glucocerebrosidase molecules was severely decreased.
在戈谢病（葡糖脑苷脂贮积症）中，葡糖脑苷脂酶缺乏导致葡糖脑苷脂的病理性蓄积，尤其是在脾脏中。因此，对正常受试者（n = 4）以及非神经病变型（n = 5）和神经病变型（n = 5）戈谢病患者脾脏中的葡糖脑苷脂酶进行了比较生化和免疫学分析。所有戈谢病患者的脾脏均显示葡糖脑苷脂酶活性明显降低。基于残余酶活性水平，或使用免疫纯化酶对动力学常数、pI或分子量形式进行测量，均无法区分戈谢病的不同表型。与从正常受试者脾脏中纯化的酶相比，从一名非神经病变型戈谢病患者脾脏中纯化至均一性的葡糖脑苷脂酶的比活性严重降低。这一发现通过一种开发用于准确评估每分子葡糖脑苷脂酶蛋白相对酶活性的免疫学方法得到了证实。该方法显示，所有研究的非神经病变型戈谢病病程患者脾脏中的残余酶每分子酶（在牛磺胆酸盐存在下测量）的葡糖脑苷脂酶活性降低了7倍以上，且这些患者脾脏中葡糖脑苷脂酶分子的浓度接近正常。免疫印迹实验的观察结果与这些发现一致。相比之下，在神经病变型戈谢病患者的脾脏中，葡糖脑苷脂酶分子的浓度严重降低。
