大鼠脂肪组织胞质溶胶中甘油三酯脂肪酶在肝素琼脂糖上的亲和层析。

Affinity chromatography on heparin-sepharose of rat adipose tissue triglyceride lipase from cytosol.

作者信息

Vanhove A, Breton M, Polonovski J

出版信息

Adv Exp Med Biol. 1978;101:127-34. doi: 10.1007/978-1-4615-9071-2_12.

Abstract

Subfractionation of the fat free homogenate of rat adipose tissue showed that a high yield of triglyceride lipase was recovered reproducibly in the microsomal supernatant fraction (cytosol) when rat epididymal fat pads were homogenized in sucrose-EDTA-Tris medium. Triglyceride lipase was bound on heparin-Sepharose. Hydrolyzing activity towards triacylglycerol was eluted as a single, sharp peak in 0.7 M NaCl, 5 mM sodium barbital and 20% glycerol (pH 7.0). The triglyceride lipase was not inhibited by 1 M NaCl and not stimulated by the presence of fresh human serum. A lipoprotein-lipase activity was demonstrable in the cytosol when adipose tissue from fed rats were used. Fasting of the animals lowered this activity.

摘要

大鼠脂肪组织无脂匀浆的亚分级分离表明，当大鼠附睾脂肪垫在蔗糖 - 乙二胺四乙酸 - 三羟甲基氨基甲烷（sucrose-EDTA-Tris）培养基中匀浆时，微粒体上清液部分（胞质溶胶）可重复性地回收高产率的甘油三酯脂肪酶。甘油三酯脂肪酶结合在肝素 - 琼脂糖上。对三酰甘油的水解活性在0.7M氯化钠、5mM巴比妥酸钠和20%甘油（pH 7.0）中以单一尖锐峰的形式洗脱。甘油三酯脂肪酶不受1M氯化钠抑制，也不受新鲜人血清存在的刺激。当使用喂食大鼠的脂肪组织时，胞质溶胶中可证明有脂蛋白脂肪酶活性。动物禁食会降低这种活性。