The predominant form of mammalian DNA topoisomerase Iin vivo has a molecular mass of 100 kDa |
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| Authors: | Scott Samuels Nobuyoshi Shimizu |
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| Institution: | (1) Department of Molecular and Cellular Biology, University of Arizona, 85721 Tucson, Arizona, USA;(2) Laboratory of Vectors and Pathogens, Rocky Mountain Laboratories, 59840 Hamilton, MT, USA;(3) Department of Molecular Biology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, 160 Tokyo, Japan |
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| Abstract: | DNA topoisomerase I was purified to apparent homogeneity from human HeLa cells as a single polypeptide with a molecular mass of 100 kDa, as assayed by both gel filtration column chromatography and SDS-polyacrylamide gel electrophoresis. No smaller forms of the enzyme were detected in the purified fraction. Therefore, smaller forms, which have been observed by other investigators, are likely to be the result of proteolysis during isolation and are not relevant to thein vivo activity of DNA topoisomerase I.Abbreviations 2-ME
2-Mercaptoethanol
- DTT
Dithiothreitol
- PMSF
Phenylmethylsulfonyl Fluoride
- SDS
Sodium Dodecyl Sulfate |
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| Keywords: | HeLa phosphorylation proteolysis purification smaller forms topoisomerase I |
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