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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  • Isolation of Deoxycytidine Kinase from Ehrlich Carcinoma Cells by Affinity Chromatography Based on a Substrate Analog, 2'-C-Cyano-2'-deoxy-1-.BETA.-D-arabinofuranosyl-N4-palmitoylcytosine.
  • Isolation of Deoxycytidine Kinase from Ehrlich Carcinoma Cells

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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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