Resolution of the diadenosine 5',5"'-P1,P4-tetraphosphate binding subunit from a multiprotein form of HeLa cell DNA polymerase alpha.
Resolution of the diadenosine 5',5"'-P1,P4-tetraphosphate binding subunit from a multiprotein form of HeLa cell DNA polymerase alpha.
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从 HeLa 细胞 DNA 聚合酶 α 的多蛋白形式中分离出二腺苷 5,5"-P1,P4-四磷酸结合亚基。
DOI：
10.1073/pnas.80.16.4931
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发表时间：
1983
影响因子：
11.1
通讯作者：
Zamecnik,P
中科院分区：
文献类型：
--
作者：
Baril,E;Bonin,P;Burstein,D;Mara,K;Zamecnik,P
一个二腺苷5',5 ' '-P1， p4 -四磷酸（Ap4A）结合亚基从高分子量（640,000）多蛋白形式的DNA聚合酶α[脱氧核苷三磷酸：DNA核苷酸转移酶（DNA-定向），EC 2.7.7.7]从HeLa细胞[DNA聚合酶α 2的Lamothe， P., Baril， B., Chi， A., Lee， L. & Baril， E. (1981) Proc. Natl.。学会科学。[j]。在纯化过程中，Ap4A结合活性与DNA聚合活性相结合。丁糖糖的疏水色谱分析了DNA聚合酶的Ap4A结合活性。Ap4A的结合活性本质上是蛋白质的，因为用蛋白酶K处理分离的结合活性会消除Ap4A的结合，但对DNase或RNase处理不敏感。用[32P]Ap4A进行光亲和标记后，用聚丙烯酰胺凝胶电泳和NaDodSO4/聚丙烯酰胺凝胶电泳测定的Ap4A结合蛋白在非变性条件下的分子量分别为92000和47000。该蛋白的结合活性对Ap4A具有高度特异性。
A diadenosine 5',5"'-P1,P4-tetraphosphate (Ap4A) binding subunit has been resolved from a high molecular weight (640,000) multiprotein form of DNA polymerase alpha [deoxynucleoside triphosphate:DNA nucleotidyltransferase (DNA-directed), EC 2.7.7.7] from HeLa cells [DNA polymerase alpha 2 of Lamothe, P., Baril, B., Chi, A., Lee, L. & Baril, E. (1981) Proc. Natl. Acad. Sci. USA 78, 4723-4727]. The Ap4A binding activity copurifies with the DNA polymerizing activity during the course of purification. Hydrophobic chromatography on butylagarose resolves the Ap4A binding activity from the DNA polymerase. The Ap4A binding activity is protein in nature since the binding of Ap4A is abolished by treatment of the isolated binding activity with proteinase K but is insensitive to treatment with DNase or RNase. The molecular weight of the Ap4A binding protein, as determined by polyacrylamide gel electrophoresis under nondenaturing conditions or by NaDodSO4/polyacrylamide gel electrophoresis after photoaffinity labeling of the protein with [32P]Ap4A is 92,000 or 47,000. The binding activity of this protein is highly specific for Ap4A.