Activation of topoisomerase II during partial purification by heparin-Sepharose chromatography

Fritz Boege; Frank Gieseler; Michaela Müller; Harald Biersack; Peter Meyer

doi:10.1016/0021-9673(92)87222-T

Activation of topoisomerase II during partial purification by heparin-Sepharose chromatography

Fritz Boege^*, Frank Gieseler, Michaela Müller, Harald Biersack, Peter Meyer

^*Corresponding author for this work

University of Würzburg

6 Citations (Scopus)

Abstract

Partial purification of topoisomerase II from small samples (10⁷-10⁸ cells) of human leukaemic cells was achieved by isolation of cell nuclei, hyper-osmotic extraction of nuclear proteins, sorption of nuclear proteins by heparin-Sepharose and elution with potassium phosphate. Similar results were obtained by gradient and batchwise elution. The catalytic activity of topoisomerase increased ca. eightfold after removal of ca. 95% of the contaminating nuclear proteins. The conserved enzymatic activity after partial purification indicates that the enzyme was not damaged. The half-life of enzymatic activity is increased by the chromatographic procedure. Owing to its high yield and technical simplicity, this could be a candidate procedure for the study of topoisomerase II in patient-derived blood samples.

Original language	English
Journal	Journal of Chromatography A
Volume	625
Issue number	1
Pages (from-to)	67-71
Number of pages	5
ISSN	0021-9673
DOIs	https://doi.org/10.1016/0021-9673(92)87222-T
Publication status	Published - 13.11.1992

Funding

This work was supported by the Wilhelm Sander-Stiftung, Grant 90.038.01, and the Deutsche For-schungsgemeinschaft,S FB 172, C9. Excellent tech- nical assistance was rendered by Michael Clark. The authors thank Professor LF. . Liu, Johns Hopkins University, Baltimore, for the gift of the anti-topoisomerase II antibody.

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@article{7476775e99394c0da0c20421ce582f91,

title = "Activation of topoisomerase II during partial purification by heparin-Sepharose chromatography",

abstract = "Partial purification of topoisomerase II from small samples (107-108 cells) of human leukaemic cells was achieved by isolation of cell nuclei, hyper-osmotic extraction of nuclear proteins, sorption of nuclear proteins by heparin-Sepharose and elution with potassium phosphate. Similar results were obtained by gradient and batchwise elution. The catalytic activity of topoisomerase increased ca. eightfold after removal of ca. 95\% of the contaminating nuclear proteins. The conserved enzymatic activity after partial purification indicates that the enzyme was not damaged. The half-life of enzymatic activity is increased by the chromatographic procedure. Owing to its high yield and technical simplicity, this could be a candidate procedure for the study of topoisomerase II in patient-derived blood samples.",

author = "Fritz Boege and Frank Gieseler and Michaela M{\"u}ller and Harald Biersack and Peter Meyer",

note = "Funding Information: This work was supported by the Wilhelm Sander-Stiftung, Grant 90.038.01, and the Deutsche For-schungsgemeinschaft,S FB 172, C9. Excellent tech- nical assistance was rendered by Michael Clark. The authors thank Professor LF. . Liu, Johns Hopkins University, Baltimore, for the gift of the anti-topoisomerase II antibody.",

year = "1992",

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day = "13",

doi = "10.1016/0021-9673(92)87222-T",

language = "English",

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T1 - Activation of topoisomerase II during partial purification by heparin-Sepharose chromatography

AU - Boege, Fritz

AU - Gieseler, Frank

AU - Müller, Michaela

AU - Biersack, Harald

AU - Meyer, Peter

N1 - Funding Information: This work was supported by the Wilhelm Sander-Stiftung, Grant 90.038.01, and the Deutsche For-schungsgemeinschaft,S FB 172, C9. Excellent tech- nical assistance was rendered by Michael Clark. The authors thank Professor LF. . Liu, Johns Hopkins University, Baltimore, for the gift of the anti-topoisomerase II antibody.

PY - 1992/11/13

Y1 - 1992/11/13

N2 - Partial purification of topoisomerase II from small samples (107-108 cells) of human leukaemic cells was achieved by isolation of cell nuclei, hyper-osmotic extraction of nuclear proteins, sorption of nuclear proteins by heparin-Sepharose and elution with potassium phosphate. Similar results were obtained by gradient and batchwise elution. The catalytic activity of topoisomerase increased ca. eightfold after removal of ca. 95% of the contaminating nuclear proteins. The conserved enzymatic activity after partial purification indicates that the enzyme was not damaged. The half-life of enzymatic activity is increased by the chromatographic procedure. Owing to its high yield and technical simplicity, this could be a candidate procedure for the study of topoisomerase II in patient-derived blood samples.

AB - Partial purification of topoisomerase II from small samples (107-108 cells) of human leukaemic cells was achieved by isolation of cell nuclei, hyper-osmotic extraction of nuclear proteins, sorption of nuclear proteins by heparin-Sepharose and elution with potassium phosphate. Similar results were obtained by gradient and batchwise elution. The catalytic activity of topoisomerase increased ca. eightfold after removal of ca. 95% of the contaminating nuclear proteins. The conserved enzymatic activity after partial purification indicates that the enzyme was not damaged. The half-life of enzymatic activity is increased by the chromatographic procedure. Owing to its high yield and technical simplicity, this could be a candidate procedure for the study of topoisomerase II in patient-derived blood samples.

UR - https://www.scopus.com/pages/publications/0026523907

U2 - 10.1016/0021-9673(92)87222-T

DO - 10.1016/0021-9673(92)87222-T

M3 - Journal articles

C2 - 12126111

AN - SCOPUS:0026523907

SN - 0021-9673

VL - 625

SP - 67

EP - 71

JO - Journal of Chromatography A

JF - Journal of Chromatography A

IS - 1

ER -

Activation of topoisomerase II during partial purification by heparin-Sepharose chromatography

Abstract

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