VISUALIZATION OF POLY(ADP-RIBOSE) SYNTHETASE ASSOCIATED WITH POLYNUCLEOSOMES BY IMMUNOELECTRON MICROSCOPY
VISUALIZATION OF POLY(ADP-RIBOSE) SYNTHETASE ASSOCIATED WITH POLYNUCLEOSOMES BY IMMUNOELECTRON MICROSCOPY
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DOI：
10.1016/0167-4889(86)90239-9
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发表时间：
1986-03-14
期刊：
影响因子：
--
通讯作者：
POIRIER, GG
中科院分区：
文献类型：
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作者：
LEDUC, Y;DEMURCIA, G;POIRIER, GG
已纯化出对聚ADP核糖合成酶具有高特异性的抗体。抗血清和非免疫血清中存在非特异性结合组蛋白的部分已通过免疫印迹和组蛋白- sepharose色谱分离得到证实。没有非特异性结合部分的抗体用小牛胸腺蛋白提取物进行Western blot分析，发现仅与116 kDa的条带反应。与纯化的拓扑异构酶I没有反应，该弱活性是用聚adp核糖合成酶制剂进行共纯化的。通过间接金标法，将特异IgG片段用于观察聚adp核糖合成酶与染色质的相互作用。这项免疫显微镜研究表明，合成酶位于多核小体的内部，并优先与核心核小体相关。
Antibodies showing a high specificity for poly(ADP ribose) synthetase have been purified. A fraction binding nonspecifically to histones present in antiserum and non-immune serum has been demonstrated by immunoblotting and separated by histone-Sepharose chromatography. The antibody without the nonspecific binding fraction was analyzed by Western blot with calf thymus protein extract and was found to react only with a band at 116 kDa. There was no reaction with purified topoisomerase I, this weak activity was copurified with poly(ADP-ribose) synthetase preparation. The specific IgG fraction has been used for the visualization of the interaction of poly(ADP-ribose) synthetase with chromatin by indirect gold-labelling. This immunomicroscopic study suggest that the synthetase is located in the inner part of polynucleosomes and would be associated preferentially with the core nucleosome.