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

doi:10.1039/c2ra21758a

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

School of Biosciences

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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 language	English
Pages (from-to)	10598-10604
Number of pages	7
Journal	RSC advances
Volume	2
Issue number	28
Early online date	7 Sept 2012
DOIs	https://doi.org/10.1039/c2ra21758a
Publication status	Published - 14 Nov 2012

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

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",

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

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doi = "10.1039/c2ra21758a",

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T1 - Efficient purification of chromatin architectural proteins

T2 - histones, HMGB proteins and FKBP3 (FKBP25) immunophilin

AU - Foulger, Larus E.

AU - Sin, Connie Goh Then

AU - Zhuang, Q.Q.

AU - Smallman, Hugh

AU - Nicholson, James M.

AU - Lambert, Stanley J.

AU - Reynolds, Colin D.

AU - Dickman, Mark J.

AU - Wood, Christopher M.

AU - Baldwin, John P.

AU - Evans, Katie

PY - 2012/11/14

Y1 - 2012/11/14

N2 - 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

AB - 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

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JO - RSC advances

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ER -

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

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