3'-PHOSPHATASE ACTIVITY IN T4 POLYNUCLEOTIDE KINASE
3'-PHOSPHATASE ACTIVITY IN T4 POLYNUCLEOTIDE KINASE
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DOI：
10.1021/bi00642a027
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发表时间：
1977-01-01
期刊：
影响因子：
2.9
通讯作者：
UHLENBECK, OC
中科院分区：
文献类型：
--
作者：
CAMERON, V;UHLENBECK, OC
[噬菌体] T4多核苷酸激酶的纯化导致活性的共纯化，该活性将在ATP不存在的情况下从多种脱氧核糖核苷酸和核糖核苷酸中特异性地去除3“-末端磷酸。这种磷酸酶活性需要Mg，最适pH为6.0，并且对脱氧核糖核苷酸比核糖核苷酸更有活性。T4多核苷酸激酶和3 ''-磷酸酶活性通过梯度洗脱柱层析法在DEAE-纤维素、磷酸纤维素和羟基磷灰石上进行共纯化。这两种活性都包含在Sephadex G-200中并在Sephadex G-200上共迁移。在pH9.2的聚丙烯酰胺凝胶电泳中，这两种活性与主要蛋白带一起共迁移。这两种活性对35 ℃下的热失活平行响应。C和ATP（仅为激酶的底物）保护两种活性免于热失活。这两种活性可能是同一个蛋白质分子的功能。
The purification of [phage] T4 polynucleotide kinase results in the copurification of an activity which will specifically remove the 3''-terminal phosphate from a variety of deoxyribonucleotides and ribonucleotides in the absence of ATP. This phosphatase activity requires Mg, has a pH optimum of 6.0, and is more active with deoxyribonucleotides than ribonucleotides. T4 polynucleotide kinase and the 3''-phosphatase activity copurify by gradient elution column chromatography on DEAE-cellulose, phosphocellulose and hydroxylapatite. The 2 activities are included in and comigrate on Sephadex G-200. Polyacrylamide gel electrophoresis at pH 9.2 results in comigration of the 2 activities together with the major protein band. The 2 activities respond in parallel to heat inactivation at 35.degree. C and ATP, a substrate for the kinase only, protects both activities from heat inactivation. The 2 activities are probably functions of the same protein molecule.