Purification, immunological and biochemical characterization of Ap4A binding protein from Xenopus laevis oocytes.
Purification, immunological and biochemical characterization of Ap4A binding protein from Xenopus laevis oocytes.
复制标题
非洲爪蟾卵母细胞 Ap4A 结合蛋白的纯化、免疫学和生化表征。
DOI：
10.1093/nar/16.7.2913
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发表时间：
1988
影响因子：
14.9
通讯作者：
L. Tarrago
中科院分区：
文献类型：
--
作者：
L. Zourgui;D. Baltz;T. Baltz;F. Oukerro;L. Tarrago
二腺苷 5',5'''-P1,P4-四磷酸 (Ap4A) 结合蛋白特异性结合 Ap4A。该蛋白质是从非洲爪蟾卵母细胞中纯化出来的，通过凝胶过滤估计分子量为 100,000。在纯化的第一阶段，发现 Ap4A 结合活性与 DNA 聚合酶 α-DNA 引物酶相关，形成异质高分子量复合物。针对纯化的 Ap4A 结合蛋白制备了单克隆抗体。该抗体部分中和 Ap4A 结合活性。使用免疫印迹技术，已表明该抗体能够识别天然或 SDS 变性的 Ap4A 结合蛋白。单克隆抗体与90,000的多肽发生免疫反应，这与凝胶色谱获得的分子量一致，表明来自非洲爪蟾卵母细胞的天然Ap4A结合蛋白可能是单体蛋白。
Diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) binding protein specifically binds Ap4A. The protein has been purified from Xenopus laevis oocytes and presents an estimated molecular weight of 100,000 by gel filtration. In the first stages of the purification, the Ap4A binding activity is found associated to DNA polymerase alpha-DNA primase, forming heterogeneous high molecular weight complexes. A monoclonal antibody has been prepared against the purified Ap4A binding protein. The antibody partially neutralizes the Ap4A binding activity. Using the immunoblot technique, it has been shown that the antibody is able to recognize either native or SDS-denatured Ap4A binding protein. The monoclonal antibody immunoreacted with a polypeptide of 90,000 which coincides with the molecular weight obtained by gel chromatography and indicates that the native Ap4A binding protein from Xenopus oocytes is probably a monomeric protein.