Purification of DNA repair protein complexes from mammalian cells.
Purification of DNA repair protein complexes from mammalian cells.
复制标题
DOI：
10.1016/j.xpro.2021.100348
复制
发表时间：
2021-03-19
期刊：
影响因子：
--
通讯作者：
Cohn MA
中科院分区：
文献类型：
--
作者：
Liang CC;Cohn MA
细胞具有多种DNA修复途径来应对各种DNA损伤。通常，DNA修复蛋白作为大的蛋白质复合物起作用。在这里，我们描述了一个协议，以纯化DNA修复蛋白复合物从哺乳动物细胞核。该方法允许纯化含有稳定以及瞬时相关蛋白质的蛋白质复合物，其随后可以通过质谱分析来鉴定。该方案可用于揭示DNA修复途径的功能和机制。有关本方案使用和执行的完整信息，请参见。从哺乳动物细胞系纯化DNA修复蛋白复合物的方案允许纯化稳定和瞬时缔合的蛋白亚基也可以纯化与DNA结合的蛋白复合物细胞具有多种DNA修复途径以解决各种DNA损伤。通常，DNA修复蛋白作为大的蛋白质复合物起作用。在这里，我们描述了一个协议，以纯化DNA修复蛋白复合物从哺乳动物细胞核。该方法允许纯化含有稳定以及瞬时缔合蛋白质的蛋白质复合物，其随后可以通过质谱分析鉴定。该方案可用于揭示DNA修复途径的功能和机制。
Cells possess multiple DNA repair pathways to tackle a variety of DNA lesions. Often, DNA repair proteins function as large protein complexes. Here, we describe a protocol to purify DNA repair protein complexes from nuclei of mammalian cells. The method permits purification of protein complexes containing stable as well as transiently associated proteins, which subsequently can be identified by mass-spectrometry analysis. This protocol can be applied to uncover the functions and mechanism of DNA repair pathways. For complete information on the use and execution of this protocol, please refer to. Protocol to purify DNA repair protein complexes from a mammalian cell line Permits purification of both stable and transiently associated protein subunits Protein complexes bound to DNA can also be purified Cells possess multiple DNA repair pathways to tackle a variety of DNA lesions. Often, DNA repair proteins function as large protein complexes. Here, we describe a protocol to purify DNA repair protein complexes from nuclei of mammalian cells. The method permits purification of protein complexes containing stable as well as transiently associated proteins, which subsequently can be identified by mass spectrometry analysis. This protocol can be applied to uncover the functions and mechanism of DNA repair pathways.
登录
查看更多内容
影响因子：
--
作者：
Aparicio, Oscar;Geisberg, Joseph V;Struhl, Kevin
通讯作者：
Struhl, Kevin
影响因子：
4.4
作者：
Singh, Abhishek A.;Schuurman, Karianne;Zwart, Wilbert
通讯作者：
Zwart, Wilbert
DOI：
10.1007/978-1-61779-376-9_7
发表时间：
2012
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Tian, Bing;Yang, Jun;Brasier, Allan R.
通讯作者：
Brasier, Allan R.
DOI：
10.1073/pnas.1902931116
发表时间：
2020-01-07
影响因子：
11.1
作者：
Mintseris, Julian;Gygi, Steven P.
通讯作者：
Gygi, Steven P.
影响因子：
2.7
作者：
Zeng, Ping-Yao;Vakoc, Christopher R.;Berger, Shelley L.
通讯作者：
Berger, Shelley L.