Sucrose gradient analysis of phospholipid-activated beta-glucosidase in type 1 and type 2 Gaucher's disease.
Sucrose gradient analysis of phospholipid-activated beta-glucosidase in type 1 and type 2 Gaucher's disease.
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1 型和 2 型戈谢病中磷脂激活的 β-葡萄糖苷酶的蔗糖梯度分析。
DOI：
10.1016/0003-9861(85)90173-0
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发表时间：
1985
影响因子：
3.9
通讯作者：
Glew,RH
中科院分区：
文献类型：
--
作者：
Garrett,KO;Prence,EM;Glew,RH
利用蔗糖密度梯度，从非神经系统（1型）和神经系统（2型）戈谢病患者的对照脾和脾脏分离的水解溶酶体β-葡萄糖苷酶的差异进行了研究。这三种酶的沉降特性不同，对磷脂酰丝氨酸和热稳定因子的活化反应也不同。对照β-葡萄糖苷酶作为一个明显的45000 - da物种沉积，其活性取决于在测定培养基中加入外源牛磺酸去氧胆酸钠。用磷脂酰丝氨酸和热稳定因子的混合物进行预孵育，将对照酶转化为快速沉积形式，在缺乏外源胆汁盐的情况下表现出相当大的活性。1型戈谢病患者脾β-葡萄糖苷酶在蔗糖梯度上的表观分子量为154,000。与对照酶一样，这种形式的活性依赖于胆汁盐。在用磷脂酰丝氨酸和热稳定因子进行预孵育后，来自1型病例的β-葡萄糖苷酶也转化为快速沉积形式，在没有牛磺酸脱氧胆酸钠的情况下比在有胆盐的情况下更活跃。来自2型戈谢病患者的脾脏β-葡萄糖苷酶在蔗糖梯度最密集的区域呈宽活性峰沉积，似乎比对照组或1型戈谢病患者的β-葡萄糖苷酶大得多。这种大型物种的活性强烈依赖于胆汁盐，并且不受磷脂酰丝氨酸和热稳定因子预孵育酶的影响。用异向盐硫氰酸钠（0.15m）将从1型戈谢病患者分离的脾脏β-葡萄糖苷酶转化为沉积较慢的物种。用NaSCN处理后，对照酶稍微深入到蔗糖梯度中。硫氰酸盐治疗对2型戈谢病脾β-葡萄糖苷酶无影响。
Using sucrose density gradients, differences in delipidated lysosomal β-glucosidase isolated from control spleen and spleen from patients with nonneurologic (type 1) and neurologic (type 2) Gaucher's disease have been examined. The three enzymes differ in sedimentation properties as well as in their responsiveness to activation by phosphatidylserine and heat-stable factor. The control β-glucosidase sedimented as an apparent 45,000-Da species whose activity was dependent upon the inclusion of exogenous sodium taurodeoxycholate in the assay medium. Preincubation with a mixture of phosphatidylserine and heat-stable factor converted the control enzyme to a faster-sedimenting form which exhibited considerable activity in the absence of exogenous bile salt. Spleen β-glucosidase from a patient with type 1 Gaucher's disease exhibited an apparent molecular weight of 154,000 on sucrose gradients. Like the control enzyme, the activity of this form was bile salt dependent. Upon preincubation with phosphatidylserine and heat-stable factor, β-glucosidase from the type 1 case was also converted to a faster-sedimenting form which was more active in the absence of sodium taurodeoxycholate than in the presence of the bile salt. Spleen β-glucosidase from the patient with type 2 Gaucher's disease sedimented as a broad peak of activity in the most dense regions of the sucrose gradients, appearing to be much larger than the β-glucosidase from either the control or the type 1 Gaucher's disease patient. The activity of this large species was strongly dependent upon bile salt, and was not affected by preincubation of the enzyme with phosphatidylserine and heat-stable factor. Using the chaotropic salt, sodium thiocyanate (0.15m), the spleen β-glucosidase isolated from the type 1 Gaucher's disease case was converted to a slower-sedimenting species. The control enzyme sedimented slightly farther into the sucrose gradients upon treatment with the NaSCN. Thiocyanate treatment had no effect on the spleen β-glucosidase isolated from the case of type 2 Gaucher's disease.