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++ +

H Uchida; Y X Chen; H Morinaga; Y Hayashi; A Matsuda; T Obata; Y Endo; T Sasaki

doi:10.1248/bpb.22.83

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++ +

Biol Pharm Bull. 1999 Jan;22(1):83-6. doi: 10.1248/bpb.22.83.

Authors

H Uchida¹, Y X Chen, H Morinaga, Y Hayashi, A Matsuda, T Obata, Y Endo, T Sasaki

Affiliation

¹ Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Fukui University, Japan.

PMID: 9989668
DOI: 10.1248/bpb.22.83

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-beta-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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Arabinonucleosides*
Carcinoma, Ehrlich Tumor / enzymology*
Chromatography, Affinity
Cytosine / analogs & derivatives*
Deoxycytidine Kinase / isolation & purification*
Deoxycytidine Kinase / metabolism
Electrophoresis, Polyacrylamide Gel
Kinetics

Substances

Arabinonucleosides
Cytosine
Deoxycytidine Kinase
sapacitabine