大肠杆菌的recBC脱氧核糖核酸酶：亚基蛋白的分离与特性鉴定及酶的重组
The recBC deoxyribonuclease of Escherichia coli: isolation and characterization of the subunit proteins and reconstitution of the enzyme.
作者信息
Lieberman R P, Oishi M
出版信息
Proc Natl Acad Sci U S A. 1974 Dec;71(12):4816-20. doi: 10.1073/pnas.71.12.4816.
Abstract
After dissociation of the E. coli recBc DNase (ATP-dependent DNase) with concentrated NaCl, two subunit proteins were isolated by ion exchange chromatography. Combination and subsequent incubation of the subunits resulted in the appearance of the original DNase. The subunit proteins, designated alpha and beta, have s(20,omega) of 4.1 S and 8.1 S, respectively. The alpha subunit possesses neither the ATP-dependent Dnase nor the DNA-dependent ATPase of the original enzyme. The beta subunit contains a low level of both enzymatic activities in a ratio markedly different from that of the original enzyme. The beta subunit complemented extracts from both recB and recC mutant strains to produce recBC DNase, while the alpha subunit did not complement either extract. These results suggest that recB and recC genes are both required for the production of beta subunit and that the recBC DNase molecule contains a protein component (alpha) that is not determined by either the recB or the recC gene.
摘要
用浓氯化钠解离大肠杆菌recBc DNA酶（ATP依赖性DNA酶）后，通过离子交换色谱法分离出两种亚基蛋白。亚基的组合及随后的孵育导致原始DNA酶的出现。这两种亚基蛋白分别命名为α和β，其沉降系数s(20,ω)分别为4.1 S和8.1 S。α亚基既不具有原始酶的ATP依赖性DNA酶活性，也不具有DNA依赖性ATP酶活性。β亚基含有低水平的这两种酶活性，其比例与原始酶明显不同。β亚基可补充recB和recC突变株的提取物以产生recBC DNA酶，而α亚基不能补充任何一种提取物。这些结果表明，recB和recC基因对于β亚基的产生都是必需的，并且recBC DNA酶分子包含一种不由recB或recC基因决定的蛋白质成分（α）。
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