Cofractionation of HMGB proteins with histone dimers

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• 作者：Qinqin Zhuang ; Hugh Smallman ; Stanley J. Lambert ; Sirirath S. Sodngam ; Colin D. Reynolds ; Katie Evans ; Mark J. Dickman ; John P. Baldwin ; Christopher M. Wood
• 关键词：Histone octamer ; Histone dimer ; Histone tetramer ; HMGB interactions ; Protein interaction domains ; Pull-down assays
• 刊名：Analytical Biochemistry
• 出版年：15 February, 2014
• 年：2014
• 卷：447
• 期：Complete
• 页码：98-106
• 全文大小：1511 K

文摘

An effective and flexible method is presented that can be used to investigate cofractionation of groups of nuclear proteins. The method was used to analyze chromatin-related proteins, of which high-mobility group B (HMGB) proteins consistently cofractionated by cation-exchange chromatography with the histone dimer (H2A-H2B). This led to the hypothesis that the two form a complex, further suggested by gel filtration, in which the HMGBs with core histones eluted as a defined high-molecular-weight peak. A necessary requirement for further studying protein interactions is that the constituents are of the highest possible purity and the pure histone dimers and tetramers used in this study were derived from pure histone octamers with their native marks. There is a growing interest in protein-protein interactions and an increasing focus on protein-interaction domains: most frequently, pull-down assays are used to examine these. The technology presented here can provide an effective system that complements pull-down assays.