Binding of HeLa DNA‐polymerase α to blue dextran: A possible artefact in molecular weight determination by gel filtration
Binding of HeLa DNA‐polymerase α to blue dextran: A possible artefact in molecular weight determination by gel filtration
复制标题
HeLa DNA聚合酶α与蓝色右旋糖酐的结合：凝胶过滤分子量测定中可能的假象
DOI：
10.1016/0014-5793(76)80166-4
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发表时间：
1976
期刊：
影响因子：
3.5
通讯作者：
P. Jeanteur
中科院分区：
文献类型：
--
作者：
C. Brissac;M. Rucheton;C. Brunei;P. Jeanteur
我们以前曾描述过通过聚（A）-琼脂糖色谱法从HeLa细胞中分离主要可溶性胞质蛋白组分[1]。用0.2 M NaCl从poly（A）柱上洗脱该蛋白质级分，SDS-丙烯酰胺凝胶电泳沿着显示含有38 000道尔顿的主要种类，并有几条次要条带。在系统地寻找这一部分中的酶的过程中，我们寻找了依赖于DNA的DNA聚合酶活性。本文件报告了这种活动的存在情况。它的物理化学和酶学特征，即（Y）DNA聚合酶的那些[2，3]，清楚地将其与上述蛋白质级分中存在的38000道尔顿的主要多肽区分开来。在本研究过程中，观察到聚合酶可以与用于测量Sephadex柱空隙体积的蓝色葡聚糖结合。本文的目的是引起人们对这一假象的注意，这一假象可能会在分子形状方面误导凝胶过滤数据的解释[4]。
We have previously described the isolation of a major soluble cytoplasmic protein fraction from HeLa cells by chromatography on poly (A)-Sepharose [1]. This protein fraction is eluted from the poly (A) column by 0.2 M NaCl and appears to contain a major species of 38 000 daltons by SDS-acrylamide gel electrophoresis along with several minor bands. In the course of a systematic search for enzymes in this fraction, we have looked for a DNA-dependent DNA-polymerase activity. The present paper reports on the presence of such an activity. Its physico-chemical and enzymological characteristics, which are those of the (Y DNA-polymerase [2, 3], clearly distinguish it from the major polypeptide of 38 000 daltons present in the above protein fraction.During the course of this study, it was observed that the polymerase could bind to the Blue Dextran used to measure the void volume of Sephadex columns. It is the purpose of this paper to bring attention to this artefact which may be misleading in the interpretation of gel filtration data in terms of molecular shape [4].