COMPARISON OF THE ABILITY OF PHOSPHOLIPIDS FROM RAT-LIVER LYSOSOMES TO RECONSTITUTE GLUCOCEREBROSIDASE
COMPARISON OF THE ABILITY OF PHOSPHOLIPIDS FROM RAT-LIVER LYSOSOMES TO RECONSTITUTE GLUCOCEREBROSIDASE
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DOI：
10.1016/0003-9861(86)90238-9
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发表时间：
1986-03-01
影响因子：
3.9
通讯作者：
HUTERER, S
中科院分区：
文献类型：
--
作者：
BASU, A;GLEW, RH;HUTERER, S
研究了大鼠肝脏葡萄糖脑苷脂酶的原位脂质激活剂。通过在蔗糖和甲曲酰胺梯度中连续等体积离心纯化大鼠肝脏溶酶体（相对于粗匀浆的 42.9 倍）。用氯仿:甲醇(2:1)提取脂质，并通过一维薄层色谱分离磷脂。溶酶体制剂的磷脂含量为0.28μmol脂质磷/mg蛋白质。磷脂酰胆碱是主要的非酸性磷脂（39.3%）。在酸性磷脂中，磷脂酰肌醇和磷脂酰丝氨酸的含量最多（分别为 12.0% 和 19.7%）。在来自戈谢脾的热稳定因子存在下，单独测试了解析的磷脂重建丁醇脱脂的大鼠肝脏葡萄糖脑苷脂酶活性的能力。单独或在热稳定因子存在下，磷脂酰丝氨酸和磷脂酰肌醇是葡萄糖脑苷脂酶最有效的激活剂。源自大鼠肝三体或兔肺巨噬细胞的双(单酰基甘油)磷酸酯在重建β-葡萄糖苷酶活性中也有效。
The in situ lipid activator of rat liver glucocerebrosidase was investigated. Rat liver lysosomes were purified (42.9-fold relative to the crude homogenate) by sequential isopycnic centrifugation in sucrose and metrizamide gradients. Lipids were extracted with chloroform:methanol (2:1) and phospholipids were separated by one-dimensional thin-layer chromatography. The phospholipid content of the lysosome preparation was 0.28 .mu.mol lipid phosphorus/mg protein. Phosphatidylcholine was present as the major nonacidic phospholipid (39.3%). Of the acidic phospholipids, phosphatidylinositol and phosphatidylserine were present in the greatest amounts (12.0 and 19.7%, respectively). The resolved phospholipids were tested separately and in the presence of a heat-stable factor from Gaucher spleen for their ability to reconstitute butanol-delipidated rat liver glucocerebrosidase activity. Alone or in the presence of the heat-stable factor, phosphatidylserine and phosphatidylinositol were the most effective activators of glucocerebrosidase. Bis(monoacylglyceryl) phosphate derived from rat liver tritosomes or rabbit lung macrophages was also effective in reconstituting .beta.-glucosidase activity.