Efficient purification of chromatin architectural proteins: histones, HMGB proteins and FKBP3 (FKBP25) immunophilin

Larus E. Foulger, Connie Goh Then Sin, Q.Q. Zhuang, Hugh Smallman, James M. Nicholson, Stanley J. Lambert, Colin D. Reynolds, Mark J. Dickman, Christopher M. Wood*, John P. Baldwin, Katie Evans

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

A two-step process of high ionic strength lysis of chicken erythrocyte cell nuclei followed by cation-exchange chromatography has separated at very high yield all the histone and HMGB (high-mobility group B) nuclear proteins, except the less-soluble histone tetramers. Surprisingly high yields of the nuclear immunophilin FKBP3 (FKBP25) and Hsp70 (heat-shock protein 70) co-fractionate with HMGB1 and HMGB3. Furthermore, these proteins can be separated by anion-exchange chromatography. The purified nuclear proteins retain their native, post-translational modification (PTM) marks, including those associated with chromatin-fibre remodelling. These marks are intimately associated with the control of the cell cycle. The methods herein are therefore of value for targeting these and other nuclear proteins for future proteomic studies in healthy and diseased cells. This journal is

Original languageEnglish
Pages (from-to)10598-10604
Number of pages7
JournalRSC advances
Volume2
Issue number28
Early online date7 Sep 2012
DOIs
Publication statusPublished - 14 Nov 2012

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    Foulger, L. E., Sin, C. G. T., Zhuang, Q. Q., Smallman, H., Nicholson, J. M., Lambert, S. J., Reynolds, C. D., Dickman, M. J., Wood, C. M., Baldwin, J. P., & Evans, K. (2012). Efficient purification of chromatin architectural proteins: histones, HMGB proteins and FKBP3 (FKBP25) immunophilin. RSC advances, 2(28), 10598-10604. https://doi.org/10.1039/c2ra21758a