TY - JOUR
T1 - Rapid purification and structural study of DNA topoisomerase I from human burkitt lymphoma raji cells
AU - Suzuki, Motoshi
AU - Takagi, Eishi
AU - Kojima, Kiyohide
AU - Izuta, Shunji
AU - Yoshida, Shonen
PY - 1989/11
Y1 - 1989/11
N2 - We have developed a rapid purification method for DNA topoisomerase I from Raji cells, a human Burkitt lymphoma cell line, using ammonium sulfate fractionation followed by chromatography on a Mono S column (FPLC, Pharmacia). By this method, the enzyme could be purified to near homogeneity within one day. Electrophoresis on sodium dodecyl sulfate polyacrylamide gel revealed that the final preparation is mainly composed of a 100-kDa protein. The major enzyme activity sedimented through a glycerol density gradient at 5.7S, accompanied with a minor peak at 8.7S. The former may correspond to the monomer of the 100-kDa polypeptide, and the latter, to its dimeric form. The gel filtration study of the crude extract revealed an active molecular species of 200 kDa, in addition to 100 kDa, and lower molecular weight forms. These results suggest that DNA topoisomerase I is largely in monomeric form, but also has a minor population of the dimeric form.
AB - We have developed a rapid purification method for DNA topoisomerase I from Raji cells, a human Burkitt lymphoma cell line, using ammonium sulfate fractionation followed by chromatography on a Mono S column (FPLC, Pharmacia). By this method, the enzyme could be purified to near homogeneity within one day. Electrophoresis on sodium dodecyl sulfate polyacrylamide gel revealed that the final preparation is mainly composed of a 100-kDa protein. The major enzyme activity sedimented through a glycerol density gradient at 5.7S, accompanied with a minor peak at 8.7S. The former may correspond to the monomer of the 100-kDa polypeptide, and the latter, to its dimeric form. The gel filtration study of the crude extract revealed an active molecular species of 200 kDa, in addition to 100 kDa, and lower molecular weight forms. These results suggest that DNA topoisomerase I is largely in monomeric form, but also has a minor population of the dimeric form.
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U2 - 10.1093/oxfordjournals.jbchem.a122926
DO - 10.1093/oxfordjournals.jbchem.a122926
M3 - Article
C2 - 2559076
AN - SCOPUS:0024807864
SN - 0021-924X
VL - 106
SP - 742
EP - 744
JO - Journal of Biochemistry
JF - Journal of Biochemistry
IS - 5
ER -