Diadenosine tetraphosphate binding protein from human HeLa cells: purification and characterization.
Diadenosine tetraphosphate binding protein from human HeLa cells: purification and characterization.
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来自人 HeLa 细胞的二腺苷四磷酸结合蛋白：纯化和表征。
作者：
Vishwanatha,JK;Wei,Z
内布拉斯加大学医学中心生物化学系，奥马哈，内布拉斯加州68198-4525，1991年8月27日接收; 1991年10月24日接收修订版摘要：普遍存在的二核苷酸P1**，/> 4-二已经提出（腺苷-5/）四磷酸（Ap 4A）参与DNA复制和细胞增殖、DNA修复、血小板聚集，和血管紧张。一种特异性结合Ap 4A的蛋白质与HeLa细胞中DNA聚合酶a（pol a2）的多蛋白形式相关。从HeLa细胞中分离纯化了Ap 4A结合蛋白。Ap 4A结合蛋白是疏水性的，通过疏水相互作用色谱法在丁基琼脂糖上从pol a2复合物中分离，随后通过色谱法在Mono-Q和Superose-12 FPLC柱上纯化至均一。Ap 4A结合活性在凝胶过滤后洗脱为单一对称峰，分子量为200 kDa。在非变性条件下进行聚丙烯酰胺凝胶电泳时，纯化的蛋白质作为200 kDa的单一蛋白质迁移。在变性条件下电泳后，结合活性被分解为45和22 kDa的两种多肽，命名为A！A2分别。A和A2可以使用同双功能交联剂辛二酸二琥珀酰亚胺酯交联。交联蛋白迁移作为一个单一的蛋白质的210 kDa的聚丙烯酰胺凝胶在变性条件下，这表明这两个多肽是一个单一的蛋白质的亚基。经Scatchard分析，该蛋白的解离常数为0.25 pM，表明Ap 4A结合蛋白与其配体具有很高的亲和力。结合活性不需要ATP。非离子去污剂Triton X-100是稳定纯化蛋白质所必需的。氨基酸组成分析表明，A1和A2是不同的。
Department of Biochemistry, University of Nebraska Medical Center, Omaha, Nebraska 68198-4525 Received August 27, 1991; Revised Manuscript Received October 24, 1991 abstract: The ubiquitous dinucleotide P1**,/> 4-di (adenosine-5/) tetraphosphate (Ap4A) has been proposed to be involved in DNA replication and cell proliferation, DNA repair, platelet aggregation, and vascular tonus. A protein binding specifically to Ap4A is associated with a multiprotein form of DNA polymerase a (pol a2) in HeLa cells. TheAp4A binding protein from HeLa cells has been purified to homogeneity starting from pol a2 complex. The Ap4A binding protein is hydrophobic and is resolved from the pol a2 complex by hydrophobic interaction chromatography on butyl-Sepharose and subsequently purified to homogeneity by chromatography on Mono-Q and Superose-12 FPLC columns. The Ap4A bindingactivity elutes as a single symmetrical peak upon gel filtration with a molecular mass of 200 kDa. Upon polyacrylamide gel electrophoresis under nondenaturing conditions, the purified protein migrates as a single protein of 200 kDa. Upon electrophoresis under denaturing conditions, the binding activity is resolved into two polypeptides of 45 and 22 kDa, designated as A! and A2, respectively. A, and A2 can be cross-linked using the homobifunctional cross-linking agent disuccinimidyl suberate. The cross-linked protein migrates as a single protein of 210 kDa on polyacrylamide gels under denaturing conditions, suggesting that these two polypeptides are subunits of a single protein. The purified protein binds Ap4Aefficiently, and by Scatchard analysis, we have determined a dissociation constant of 0.25 pM, indicating high affinity of Ap4A binding protein to its ligand. ATP is not required for the binding activity. The nonionic detergent Triton X-100 is necessary for stabilizing the purified protein. Amino acid compositionanalysis indicates that A, and A2 are distinct.
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DOI：
10.1016/s0021-9258(18)61070-1
发表时间：
1987-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
P. Matsudaira
通讯作者：
P. Matsudaira
非洲爪蟾卵母细胞 Ap4A 结合蛋白的纯化、免疫学和生化表征。
影响因子：
14.9
作者：
L. Zourgui;D. Baltz;T. Baltz;F. Oukerro;L. Tarrago
通讯作者：
L. Tarrago
通过将二腺苷 5,5-P1, P4-四磷酸 (Ap4A) 显微注射到非洲爪蟾卵母细胞中刺激 DNA 合成。
影响因子：
2.7
作者：
L. Zourgui;D. Tharaud;A. Solari;S. Litvak;L. Tarrago
通讯作者：
L. Tarrago
小牛胸腺的腺苷(5)四磷酸(5)腺苷结合蛋白。
DOI：
--
发表时间：
1984
期刊：
European Journal of Biochemistry
影响因子：
--
作者：
E. Rapaport;L. Feldman
通讯作者：
L. Feldman
细胞内腺苷 (5) 四磷酸 (5) 腺苷的急剧增加与真核细胞 DNA 合成的开始相关。
DOI：
10.1111/j.1432-1033.1984.tb07897.x
发表时间：
1984
期刊：
European journal of biochemistry
影响因子：
--
作者：
C. Weinmann;A. Hedl;I. Grummt;W. Albert;F. Ferdinand;R. Friis;G. Pierron;W. Moll;F. Grummt
通讯作者：
F. Grummt