Deoxycytidine kinase: properties of the enzyme from human leukemic granulocytes

C. N. Coleman, R. G. Stoller, J. C. Drake, B. A. Chabner

Research output: Contribution to journalArticle

Abstract

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 7.8μM for CdR and 25.6 μM for ara C, and optimal activity with ATP and GTP as phosphate donors. Ara C phosphorylation was strongly inhibited by CdR (K(i)=0.17μM) and dCTP (K(i)=7.3μM) and was weakly inhibited by ara CTP (K(i)=0.13mM). 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.

Original languageEnglish (US)
Pages (from-to)791-803
Number of pages13
JournalBlood
Volume46
Issue number5
StatePublished - 1975
Externally publishedYes

Fingerprint

Deoxycytidine Kinase
Cytarabine
Granulocytes
Phosphotransferases
Cytidine Deaminase
Enzymes
Arabinofuranosylcytosine Triphosphate
Uridine Kinase
Azacitidine
Granulocyte Precursor Cells
Ficoll
Deoxycytidine
Phosphorylation
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Chromatography
Guanosine Triphosphate
Cell Extracts
Complex Mixtures
Sedimentation
Metabolism

ASJC Scopus subject areas

  • Hematology

Cite this

Coleman, C. N., Stoller, R. G., Drake, J. C., & Chabner, B. A. (1975). Deoxycytidine kinase: properties of the enzyme from human leukemic granulocytes. Blood, 46(5), 791-803.

Deoxycytidine kinase : properties of the enzyme from human leukemic granulocytes. / Coleman, C. N.; Stoller, R. G.; Drake, J. C.; Chabner, B. A.

In: Blood, Vol. 46, No. 5, 1975, p. 791-803.

Research output: Contribution to journalArticle

Coleman, CN, Stoller, RG, Drake, JC & Chabner, BA 1975, 'Deoxycytidine kinase: properties of the enzyme from human leukemic granulocytes', Blood, vol. 46, no. 5, pp. 791-803.
Coleman CN, Stoller RG, Drake JC, Chabner BA. Deoxycytidine kinase: properties of the enzyme from human leukemic granulocytes. Blood. 1975;46(5):791-803.
Coleman, C. N. ; Stoller, R. G. ; Drake, J. C. ; Chabner, B. A. / Deoxycytidine kinase : properties of the enzyme from human leukemic granulocytes. In: Blood. 1975 ; Vol. 46, No. 5. pp. 791-803.
@article{1a6ee6f9a239483ca55528ff397e6439,
title = "Deoxycytidine kinase: properties of the enzyme from human leukemic granulocytes",
abstract = "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 7.8μM for CdR and 25.6 μM for ara C, and optimal activity with ATP and GTP as phosphate donors. Ara C phosphorylation was strongly inhibited by CdR (K(i)=0.17μM) and dCTP (K(i)=7.3μM) and was weakly inhibited by ara CTP (K(i)=0.13mM). 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.",
author = "Coleman, {C. N.} and Stoller, {R. G.} and Drake, {J. C.} and Chabner, {B. A.}",
year = "1975",
language = "English (US)",
volume = "46",
pages = "791--803",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "5",

}

TY - JOUR

T1 - Deoxycytidine kinase

T2 - properties of the enzyme from human leukemic granulocytes

AU - Coleman, C. N.

AU - Stoller, R. G.

AU - Drake, J. C.

AU - Chabner, B. A.

PY - 1975

Y1 - 1975

N2 - 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 7.8μM for CdR and 25.6 μM for ara C, and optimal activity with ATP and GTP as phosphate donors. Ara C phosphorylation was strongly inhibited by CdR (K(i)=0.17μM) and dCTP (K(i)=7.3μM) and was weakly inhibited by ara CTP (K(i)=0.13mM). 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.

AB - 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 7.8μM for CdR and 25.6 μM for ara C, and optimal activity with ATP and GTP as phosphate donors. Ara C phosphorylation was strongly inhibited by CdR (K(i)=0.17μM) and dCTP (K(i)=7.3μM) and was weakly inhibited by ara CTP (K(i)=0.13mM). 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.

UR - http://www.scopus.com/inward/record.url?scp=0016824038&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0016824038&partnerID=8YFLogxK

M3 - Article

C2 - 51655

AN - SCOPUS:0016824038

VL - 46

SP - 791

EP - 803

JO - Blood

JF - Blood

SN - 0006-4971

IS - 5

ER -