Exploring DNA-binding proteins with in vivo chemical cross-linking and mass spectrometry.
Exploring DNA-binding proteins with in vivo chemical cross-linking and mass spectrometry.
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DOI：
10.1021/pr8009319
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发表时间：
2009-04
影响因子：
4.4
通讯作者：
Wang, Yinsheng
中科院分区：
文献类型：
--
作者：
Qiu, Haibo;Wang, Yinsheng
关键词：
DNA结合蛋白是所有物种蛋白质组的重要组成部分，在转录、DNA复制、重组、修复等与DNA相关的活动中起着至关重要的作用。尽管许多DNA结合蛋白已经被鉴定出来，但许多参与基因调控和DNA修复的蛋白可能仍然未知，因为它们与DNA的动态和/或弱相互作用。在本报告中，我们描述了一种利用体内甲醛交联和LC-MS/MS综合鉴定dna结合蛋白的方法。dna结合蛋白可以通过分离dna -蛋白复合物纯化，并通过逆转交联从复合物中释放出来。通过这种方法，我们能够鉴定出一百多种DNA结合蛋白，如参与转录、基因调控、DNA复制和修复的蛋白，以及大量可能与DNA和DNA结合蛋白相关的蛋白。该方法应普遍适用于其他核酸结合蛋白的研究，在蛋白质组学水平上对基因调控、DNA损伤反应与修复以及其他许多关键生物过程的综合研究具有很大的潜力。
DNA-binding proteins are very important constituents of proteomes of all species and play crucial roles in transcription, DNA replication, recombination, repair and other activities associated with DNA. Although a number of DNA-binding proteins have been identified, many proteins involved in gene regulation and DNA repair are likely still unknown because of their dynamic and/or weak interactions with DNA. In this report, we described an approach for the comprehensive identification of DNA-binding proteins with in vivo formaldehyde cross-linking and LC-MS/MS. DNA-binding proteins could be purified via the isolation of DNA-protein complexes and released from the complexes by reversing the cross-linking. By using this method, we were able to identify more than one hundred DNA-binding proteins, such as proteins involved in transcription, gene regulation, DNA replication and repair, and a large number of proteins which are potentially associated with DNA and DNA-binding proteins. This method should be generally applicable to the investigation of other nucleic acid-binding proteins, and hold great potential in the comprehensive study of gene regulation, DNA damage response and repair, as well as many other critical biological processes at proteomic level.
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影响因子：
14.9
作者：
McCarthy FM;Bridges SM;Wang N;Magee GB;Williams WP;Luthe DS;Burgess SC
通讯作者：
Burgess SC
DOI：
10.1006/meth.1999.0879
发表时间：
1999-11-01
期刊：
METHODS-A COMPANION TO METHODS IN ENZYMOLOGY
影响因子：
--
作者：
Kuo, MH;Allis, CD
通讯作者：
Allis, CD
影响因子：
2.9
作者：
Malmström, J;Larsen, K;Marko-Varga, G
通讯作者：
Marko-Varga, G
影响因子：
4.8
作者：
Barker, S;Weinfeld, M;Murray, D
通讯作者：
Murray, D
影响因子：
3.4
作者：
Fuentes, M;Mateo, C;Fernandez-Lafuente, R
通讯作者：
Fernandez-Lafuente, R