Simultaneous purification of DNA topoisomerase I and II from eukaryotic cells.
We have developed a procedure for the simultaneous purification of DNA topoisomerase I and II from calf thymus. Both enzymes were first extracted from isolated nucleoprotein complexes. After batchwise chromatography on hydroxylapatite the two enzyme activities were separated on a FPLC phenylsuperose column. The enzymes were further purified by a second chromatography on phenylsepharose (topo I) or FPLC Mono Q (topo II). The purification can be finished within three days, yielding 0.5-1.0 mg quantities of homogeneous, enzymatic active preparations of the two proteins from 200 g of starting material.
Strausfeld
U
U
Universität Konstanz, Fakultät für Biologie, FRG.
Richter
A
A
D000818:Animals; D002417:Cattle; D002477:Cells / Q000201:enzymology*; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004796:Clinical Enzyme Tests; D004264:DNA Topoisomerases, Type I / Q000302:isolation & purification*; D004250:DNA Topoisomerases, Type II / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D005057:Eukaryotic Cells / Q000201:enzymology*; D009698:Nucleoproteins / Q000032:analysis; D013950:Thymus Gland / Q000032:analysis
D009698:Nucleoproteins; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II