Cofractionation of HMGB proteins with histone dimers
| Autor: | Hugh Smallman, Katie Evans, John P. Baldwin, Christopher M. Wood, S.J. Lambert, Qinqin Zhuang, Mark J. Dickman, Colin D. Reynolds, Sirirath Sodngam |
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| Rok vydání: | 2014 |
| Předmět: |
Cell Nucleus
Erythrocytes Size-exclusion chromatography Biophysics Cell Biology Chemical Fractionation Biology Chromatography Ion Exchange Biochemistry Protein–protein interaction Histones Histone Histone H1 HMGB Proteins Histone H2A Histone methylation biology.protein Animals Histone code Histone octamer Protein Multimerization Protein Structure Quaternary Chickens Molecular Biology |
| Zdroj: | Analytical Biochemistry. 447:98-106 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2013.11.001 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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