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  • 标题:Isolation of Deoxycytidine Kinase from Ehrlich Carcinoma Cells by Affinity Chromatography Based on a Substrate Analog, 2'-C-Cyano-2'-deoxy-1-β-D-arabinofuranosyl-N4-palmitoylcytosine
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  • 作者:Hiroyuki UCHIDA ; Yi-Xin CHEN ; Hidetaka MORINAGA
  • 期刊名称:Biological and Pharmaceutical Bulletin
  • 印刷版ISSN:0918-6158
  • 电子版ISSN:1347-5215
  • 出版年度:1999
  • 卷号:22
  • 期号:1
  • 页码:83-86
  • DOI:10.1248/bpb.22.83
  • 出版社:The Pharmaceutical Society of Japan
  • 摘要:Deoxycytidine kinase from Ehrlich carcinoma cells was purified 10400-fold by ammonium sulfate fractionation and affinity chromatography using Sepharose 4B coupled to 2'-C-cyano-2'-deoxy-1-β-D-arabinofuranosyl-N4-palmitoylcytosine, with a yield of 45%. The purified enzyme preparation showed a single major band with a molecular weight of 32000 on SDS-PAGE. The enzyme phosphorylated deoxyadenosine, deoxyguanosine, cytidine, and several deoxycytidine analogues as well as deoxycytidine. Also, the kinetic parameters of the enzyme for the substrates were estimated.
  • 关键词:deoxycytidine kinase;purification;affinity chromatography;Ehrilich carcinoma cell;kinetic parameter
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