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

研究成果: Article

1 引用 (Scopus)

抄録

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.

元の言語English
ページ(範囲)742-744
ページ数3
ジャーナルJournal of Biochemistry
106
発行部数5
DOI
出版物ステータスPublished - 01-01-1989
外部発表Yes

Fingerprint

Type I DNA Topoisomerase
Burkitt Lymphoma
Purification
Distillation columns
Ammonium Sulfate
Enzyme activity
Enzymes
Fractionation
Chromatography
Electrophoresis
Complex Mixtures
Sodium Dodecyl Sulfate
Glycerol
Gel Chromatography
Monomers
Molecular Weight
Gels
Molecular weight
Cells
Cell Line

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

これを引用

Suzuki, Motoshi ; Takagi, Eishi ; Kojima, Kiyohide ; Izuta, Shunji ; Yoshida, Shonen. / Rapid purification and structural study of DNA topoisomerase I from human burkitt lymphoma raji cells. :: Journal of Biochemistry. 1989 ; 巻 106, 番号 5. pp. 742-744.
@article{6de84c0ab9a84f90a7a69606d29cedde,
title = "Rapid purification and structural study of DNA topoisomerase I from human burkitt lymphoma raji cells",
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.",
author = "Motoshi Suzuki and Eishi Takagi and Kiyohide Kojima and Shunji Izuta and Shonen Yoshida",
year = "1989",
month = "1",
day = "1",
doi = "10.1093/oxfordjournals.jbchem.a122926",
language = "English",
volume = "106",
pages = "742--744",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "5",

}

Rapid purification and structural study of DNA topoisomerase I from human burkitt lymphoma raji cells. / Suzuki, Motoshi; Takagi, Eishi; Kojima, Kiyohide; Izuta, Shunji; Yoshida, Shonen.

:: Journal of Biochemistry, 巻 106, 番号 5, 01.01.1989, p. 742-744.

研究成果: Article

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 - 1989/1/1

Y1 - 1989/1/1

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.

UR - http://www.scopus.com/inward/record.url?scp=0024807864&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024807864&partnerID=8YFLogxK

U2 - 10.1093/oxfordjournals.jbchem.a122926

DO - 10.1093/oxfordjournals.jbchem.a122926

M3 - Article

C2 - 2559076

AN - SCOPUS:0024807864

VL - 106

SP - 742

EP - 744

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 5

ER -