Rapid purification and structural study of DNA topoisomerase I from human burkitt lymphoma raji cells

Motoshi Suzuki, Eishi Takagi, Kiyohide Kojima, Shunji Izuta, Shonen Yoshida

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)742-744
Number of pages3
JournalJournal of Biochemistry
Volume106
Issue number5
DOIs
Publication statusPublished - 11-1989
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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