DEOXYRIBONUCLEASE-II PURIFIED FROM THE ISOLATED LYSOSOMES OF PORCINE SPLEEN AND FROM PORCINE LIVER HOMOGENATES - COMPARISON WITH DEOXYRIBONUCLEASE-II PURIFIED FROM PORCINE SPLEEN HOMOGENATES
DEOXYRIBONUCLEASE-II PURIFIED FROM THE ISOLATED LYSOSOMES OF PORCINE SPLEEN AND FROM PORCINE LIVER HOMOGENATES - COMPARISON WITH DEOXYRIBONUCLEASE-II PURIFIED FROM PORCINE SPLEEN HOMOGENATES
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DOI：
10.1016/0167-4781(89)90124-3
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发表时间：
1989-01-23
期刊：
影响因子：
--
通讯作者：
LU, KS
中科院分区：
文献类型：
--
作者：
LIAO, TH;LIAO, WC;LU, KS
猪脾DNA酶II（EC 3.1.22.1）是表征最好的DNA酶II之一，其位于溶酶体中的亚细胞内，因为该酶与两种溶酶体标记酶组织蛋白酶D和酸性磷酸酶共沉积。从猪脾分离的溶酶体中纯化的DNase II的理化性质，包括亚基结构，对碘乙酸灭活的敏感性，天然分子量和色谱行为，与从整个猪脾匀浆中纯化的相同的酶没有区别。也可以用0.05 M H2SO 4或0.1 M NaCl从猪肝中提取DNA酶II，并通过一系列柱层析从任一提取物中纯化。来自任一提取物的纯化的肝DNA酶II具有相同的亚基结构（α-链，35，000和β先生-链，Mr 10，000）作为猪脾的纯化DNA酶II。两种肝脏提取物以及脾和胃粘膜提取物含有DNase II，其在Sephadex G-100凝胶过滤、非变性条件下酸性聚丙烯酰胺凝胶电泳和等电聚焦上具有非常相似的性质。这些数据有力地表明，对于同一种动物，不同组织中的DNase II活性与相同结构的蛋白质分子相关。
Porcine spleen DNase II (EC 3.1.22.1), one of the best-characterized DNase II, is subcellularly located in lysosomes because the enzyme is co-sedimented with two of the lysosomal marker enzymes, cathepsin D and acid phosphatase. The physicochemical properties, including the subunit structure, sensitivity to iodoacetate inactivation, native molecular weight and chromatographic behavior, of the DNase II purified from the isolated lysosomes of porcine spleen are indistinguishable from those of the same enzyme purified from the whole porcine spleen homogenate. DNase II can also be extracted from porcine liver with 0.05 M H2SO4 or 0.1 M NaCl and purified from either extract by a series of column chromatographies. The purified liver DNase II from either extract has the same subunit structure (.alpha.-chain, Mr 35,000 and .beta.-chain, Mr 10,000) as the purified DNase II of porcine spleen. The two liver extracts as well as the extracts of spleen and gastric mucosa contain DNase II with very similar properties on Sephadex G-100 gel filtration, on acid polyacrylamide gel electrophoresis under non-denaturing conditions, and on isoelectric focusing. The data strongly suggest that, for the same species of animal, the DNase II activities in various tissues are associated with protein molecules of identical structure.