Partial purification and identification of hormone-sensitive lipase from chicken adipose tissue
Partial purification and identification of hormone-sensitive lipase from chicken adipose tissue
复制标题
DOI：
10.1006/bbrc.1997.6923
复制
发表时间：
1997-07-09
影响因子：
3.1
通讯作者：
Holm, C
中科院分区：
文献类型：
--
作者：
Anthonsen, MW;Degerman, E;Holm, C
与来自大鼠和人脂肪组织的La HSL相比，来自鸡脂肪组织的HSL在与cAMP依赖性蛋白激酶（cAMP-PK）磷酸化后表现出显著的活化。为了表征鸡HSL酶，使用pH 5.2沉淀和阴离子交换色谱法从鸡脂肪组织匀浆中纯化3500倍。Western blot分析表明，纯化的鸡HSL蛋白分子量均为86 kDa。HSL二酰基甘油脂肪酶活性抑制98%后，与抗大鼠ASE抗血清孵育，鸡HSE的比活性估计是大约相同的大鼠酶。此外，86 kDa多肽被GAMP-PR磷酸化至与重组大鼠酶的化学计量大约相同。因此，我们的结果表明，来自鸡脂肪组织的HSL在大小和比活性上与来自哺乳动物物种的HSL相当，而不是如先前报道的具有低1000倍比活性的较小的42 kDa多肽（Berglund，L.，Khoo，J.C.，詹森博士，和斯坦伯格，D.，1980 J.Biol.Chem.255，5420-5428）。(C)北京：科学出版社.
HSL from chicken adipose tissue exhibits remarkable activation upon phosphorylation with cAMP-dependent protein kinase (cAMP-PK) compared La HSL from rat and human adipose tissue. In order to characterize the chicken HSL enzyme, it was purified 3500 fold from a chicken adipose tissue homogenate using pH 5.2 precipitation and anion-exchange chromatography. The purified chicken HSL was identified as all 86 kDa protein using Western blot analysis. The HSL diacylglycerol lipase activity was inhibited by 98% upon incubation with anti-rat ASE antiserum, and the specific activity of chicken HSE was estimated to be approximately the same as for the rat enzyme. Furthermore, the 86 kDa polypeptide was phosphorylated by GAMP-PR to about the same stoichiometry as for the recombinant rat enzyme. Hence, our results demonstrate that HSL from chicken adipose tissue is comparable in size and specific activity to HSL from mammalian species, and not a smaller 42 kDa polypeptide with 1000-fold lower specific activity as previously reported (Berglund, L., Khoo, J. C., Jensen, D., and Steinberg, D., 1980 J. Biol. Chem. 255, 5420-5428). (C) 1997 Academic Press.