大鼠肝脏微粒体的甘油单酯脂肪酶、甘油三酯脂肪酶和磷脂酶A是不同的蛋白质实体吗？
Are monoglyceride-lipase, triglyceride-lipase and phospholipase A of rat liver microsomes distinct protein entities?
作者信息
Colbeau A, Ngo-Tri H, Chabert J, Vignais P M
出版信息
Biochimie. 1977;59(5-6):517-26. doi: 10.1016/s0300-9084(77)80057-6.
PMID:889935
Abstract
- Different extraction and purification techniques were employed for the separation of MG-lipase, TG-lipase and phospholipase A from rat liver microsomes. 2. Up to 60 per cent of the microsomal content of TG-lipase and phospholipase could be extracted with 1 M KCl or NaCl. MG-lipase was extracted more readily by detergents (e.g. emulphogen). 3. MG-lipase is more resistant to detergent and heat inactivation than TG-lipase and phospholipase A. It is retained, using the technique of affinity chromatography, on a column of CH-Sepharose coupled to monooleoylglycerol. In addition, MG-lipase was separated from TG-lipase by electrofocusing. 4. TG-lipase and phospholipase A were partially separated by gel filtration on Sephadex G-200 in the presence of 1 mM dithiothreitol and by chromatography on CH-Sepharose 4b. 5. On the basis of the present extraction and purification studies, it is concluded that mg-lipase is an enzyme protein distinct from TG-lipase and phospholipase A.
摘要
- 采用不同的提取和纯化技术从大鼠肝脏微粒体中分离单甘酯脂肪酶、甘油三酯脂肪酶和磷脂酶A。2. 高达60%的甘油三酯脂肪酶和磷脂酶的微粒体含量可用1M氯化钾或氯化钠提取。单甘酯脂肪酶更容易被去污剂（如乳化剂）提取。3. 单甘酯脂肪酶比甘油三酯脂肪酶和磷脂酶A对去污剂和热失活更具抗性。使用亲和色谱技术，它保留在偶联单油酰甘油的CH-琼脂糖柱上。此外，通过等电聚焦将单甘酯脂肪酶与甘油三酯脂肪酶分离。4. 在1mM二硫苏糖醇存在下，通过Sephadex G-200凝胶过滤和CH-琼脂糖4b柱色谱法将甘油三酯脂肪酶和磷脂酶A部分分离。5. 根据目前的提取和纯化研究，得出结论：单甘酯脂肪酶是一种与甘油三酯脂肪酶和磷脂酶A不同的酶蛋白。
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