Activation of topoisomerase II during partial purification by heparin-Sepharose chromatography.
Partial purification of topoisomerase II from small samples (10(7)-10(8) 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 calatylic 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.
Boege
F
F
Medizinische Poliklinik der Universität Würzburg, Germany.
Gieseler
F
F
Müller
M
M
Biersack
H
H
Meyer
P
P
D002853:Chromatography, Liquid / Q000379:methods*; D004250:DNA Topoisomerases, Type II / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D018922:HL-60 Cells; D006493:Heparin; D006801:Humans; D012685:Sepharose
D006493:Heparin; D012685:Sepharose; D004250:DNA Topoisomerases, Type II