TY - JOUR
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 - http://www.scopus.com/inward/record.url?scp=0026523907&partnerID=8YFLogxK
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 -
