Deoxycytidine kinase: properties of the enzyme from human leukemic granulocytes.
Deoxycytidine kinase, which phosphorylates deoxycytidine (CdR) and its analog, cytosine arabinoside (ara-C), has been purified 71-fold from human leukemic cells. Biochemical properties of the partially purified enzyme included a molecular weight of 68,000, Kms of 7.8 muM for CdR and 25.6 muM for ara-C, and optimal activity with ATP and GTP as phosphate donors. Ara-C phosphorylation was strongly inhibited by CdR (Ki = 0.17 muM) and dCTP (Ki = 7.3 muM) and was weakly inhibited by ara-CTP (Ki = 0.13 mM). Purification by calcium phosphate gel elution and DEAE chromatography effectively separated this enzyme from cytidine deaminase, which deaminates both CdR and ara-C, and from uridine-cytidine kinase, the enzyme which phosphorylates 5-azacytidine. CdR kinase activity was found to decrease and cytidine deaminase to increase with maturation of normal and leukemic granulocytes. Myeloblasts purified by Ficoll sedimentation revealed an average kinase activity of 15.4 U/mg protein in acute myelocytic leukemia and 12.3 U/mg protein in blastic crisis of chronic myelocytic leukemia (CML). The average ratio of CdR kinase to deaminase activity in crude cell extracts varied from 0.197 in AML and 0.089 in blastic crisis to 0.0004 in normal granulocytes, reflecting the changes which take place with cellular maturation. The absolute levels of kinase and deaminase and the ratio of these two enzymes varied considerably among patients with AML, indicating that quantitative differences may be found in the metabolism of CdR and its analogs in leukemic cells.
D001374:Azacitidine / Q000494:pharmacology; D001798:Blood Proteins / Q000032:analysis; D002469:Cell Separation; D002474:Cell-Free System; D003561:Cytarabine / Q000494:pharmacology; D003564:Cytidine Deaminase / Q000097:blood; D003841:Deoxycytidine; D006098:Granulocytes / Q000201:enzymology*; D006801:Humans; D007938:Leukemia / Q000097:blood*; D007951:Leukemia, Myeloid / Q000097:blood; D015470:Leukemia, Myeloid, Acute / Q000097:blood; D007962:Leukocytes / Q000201:enzymology*; D008970:Molecular Weight; D010710:Phosphates / Q000378:metabolism; D010770:Phosphotransferases / Q000097:blood*; D014020:Tissue Extracts; D014541:Uridine Kinase / Q000097:blood
D001798:Blood Proteins; D010710:Phosphates; D014020:Tissue Extracts; D003561:Cytarabine; D003841:Deoxycytidine; D010770:Phosphotransferases; D014541:Uridine Kinase; D003564:Cytidine Deaminase; D001374:Azacitidine