Samuels D S, Shimizu N
Department of Molecular and Cellular Biology, University of Arizona, Tucson 85721.
Mol Biol Rep. 1994 Mar;19(2):99-103. doi: 10.1007/BF00997154.
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 the in vivo activity of DNA topoisomerase I.
通过凝胶过滤柱色谱法和SDS-聚丙烯酰胺凝胶电泳测定,从人宫颈癌细胞系HeLa细胞中纯化出的DNA拓扑异构酶I呈现为单一多肽,分子量为100 kDa,达到了表观均一性。在纯化组分中未检测到该酶的较小形式。因此,其他研究者所观察到的较小形式可能是分离过程中蛋白水解的结果,与DNA拓扑异构酶I的体内活性无关。