Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
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
Hiroyuki UCHIDAYi-Xin CHENHidetaka MORINAGAYuko HAYASHIAkira MATSUDATohru OBATAYoshio ENDOTakuma SASAKI
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1999 Volume 22 Issue 1 Pages 83-86

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Abstract

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.

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© The Pharmaceutical Society of Japan
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