PURIFICATION OF AN ENDONUCLEASE FROM VENOM OF BOTHROPS ATROX
PURIFICATION OF AN ENDONUCLEASE FROM VENOM OF BOTHROPS ATROX
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DOI：
10.1021/bi00908a016
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发表时间：
1962-01-01
期刊：
影响因子：
2.9
通讯作者：
LASKOWSKI, M
中科院分区：
文献类型：
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作者：
GEORGATSOS, JG;LASKOWSKI, M
磷酸二酯酶广泛用于核苷酸序列的建立，其可能的污染物性质值得研究。从42%的丙酮沉淀物（制备毒液磷酸二酯酶的副产物）中纯化了一种肉毒杆菌毒液的核酸酶，纯度约为1000倍。由此获得的酶制剂可以以相似的速率分裂核糖核酸和脱氧核糖核酸。该酶在pH 5.0时具有最佳活性，不需要镁。它作为一种核酸内切酶作用于DNA，并主要产生三或更高的寡核苷酸，所有这些核苷酸终止于3[图像]-单酯化磷酸。在消化的早期阶段，de-Gp-Gp1是最易受影响的键。随着酶解的进行，对相邻碱基的特异性降低，底物链的长度变得更加显著。经过彻底的消化后，得到了所有四个碱基在末端位置几乎随机分布的片段。
Since phosphodiesterase is widely used for establishing nucleotide sequence, the properties of its possible contaminants deserve study. A nuclease of the venom of Bothrops atrox has been purified approximately 1000-fold from the 42% acetone precipitate, which is a by-product of the preparation of venom phosphodiesterase. The preparation of enzyme thus obtained can split both ribo- and deoxyribo-nucleic acids at a similar rate. The enzyme has an optimal activity at pH 5.0 and requires no magnesium. It acts on DNA as an endonuclease, and produces predominantly tri- or higher oligonucleotides all of which terminate in 3[image]-monoesterified phosphate. At the early stages of digestion de-Gp-Gp1 is the most susceptible bond. As the digestion progresses the specificity in respect to the adjacent bases decreases, and the length of the substrate chain becomes more significant. After an exhaustive digestion fragments are obtained in which all four bases in terminal positions occur in an almost random distribution.