Pyrimidine salvage enzymes in human tonsil lymphocytes

II. Purification and properties of deoxycytidine kinase.

K. Szyfter, M. Sasvári, T. Spasokukotskaja, F. Antoni, M. Staub

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Human tonsillar lymphocytes incorporate about 7-10 times more [3H]-deoxy-thymidine than [3H]-deoxycytidine at the same extracellular nucleoside concentration, and both incorporations could be inhibited by 10(-5) M arabinosyl-cytosine to 99%. On the other hand, the crude extract of these cells had about 10 times more deoxycytidine kinase than deoxythymidine kinase specific activity, as it was reported previously as well. Therefore, the differences in the labelling of these cells by [3H]-deoxycytidine and [3H]-thymidine cannot be simple due to the different levels of phosphorylating enzymes in the cells. The deoxycytidine kinase of human tonsillar lymphocytes was purified 38.6-fold by DEAE-Sephadex chromatography, and some kinetic and regulatory properties of the purified enzyme were investigated. Deoxycytidine kinase was eluted as a single peak from DEAE-Sephadex column and also from Sephadex G-100 column indicating a molecular weight of about 60 000; no isoenzymes could be detected. During the purification procedure an increase of the enzyme activity was observed suggesting the inhibition of the enzyme in the cells. An apparent Km of 13 microM for deoxycytidine and Ki of 55 microM for arabinosyl-cytosine were found for the tonsillar deoxycytidine kinase. The enzyme was strongly inhibited by dCTP, but not by the other deoxyribonucleoside triphosphates.

Original languageEnglish
Pages (from-to)173-182
Number of pages10
JournalActa Biochimica et Biophysica Academiae Scientiarum Hungaricae
Volume20
Issue number3-4
Publication statusPublished - 1985

Fingerprint

Deoxycytidine Kinase
Palatine Tonsil
Deoxycytidine
Lymphocytes
Enzymes
DEAE-Dextran
Cytosine
Thymidine
Deoxyribonucleosides
Thymidine Kinase
Complex Mixtures
Nucleosides
Isoenzymes
Chromatography
Molecular Weight
pyrimidine

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Pyrimidine salvage enzymes in human tonsil lymphocytes : II. Purification and properties of deoxycytidine kinase. / Szyfter, K.; Sasvári, M.; Spasokukotskaja, T.; Antoni, F.; Staub, M.

In: Acta Biochimica et Biophysica Academiae Scientiarum Hungaricae, Vol. 20, No. 3-4, 1985, p. 173-182.

Research output: Contribution to journalArticle

@article{7232d33352d041c8b379a3341c2fc7af,
title = "Pyrimidine salvage enzymes in human tonsil lymphocytes: II. Purification and properties of deoxycytidine kinase.",
abstract = "Human tonsillar lymphocytes incorporate about 7-10 times more [3H]-deoxy-thymidine than [3H]-deoxycytidine at the same extracellular nucleoside concentration, and both incorporations could be inhibited by 10(-5) M arabinosyl-cytosine to 99{\%}. On the other hand, the crude extract of these cells had about 10 times more deoxycytidine kinase than deoxythymidine kinase specific activity, as it was reported previously as well. Therefore, the differences in the labelling of these cells by [3H]-deoxycytidine and [3H]-thymidine cannot be simple due to the different levels of phosphorylating enzymes in the cells. The deoxycytidine kinase of human tonsillar lymphocytes was purified 38.6-fold by DEAE-Sephadex chromatography, and some kinetic and regulatory properties of the purified enzyme were investigated. Deoxycytidine kinase was eluted as a single peak from DEAE-Sephadex column and also from Sephadex G-100 column indicating a molecular weight of about 60 000; no isoenzymes could be detected. During the purification procedure an increase of the enzyme activity was observed suggesting the inhibition of the enzyme in the cells. An apparent Km of 13 microM for deoxycytidine and Ki of 55 microM for arabinosyl-cytosine were found for the tonsillar deoxycytidine kinase. The enzyme was strongly inhibited by dCTP, but not by the other deoxyribonucleoside triphosphates.",
author = "K. Szyfter and M. Sasv{\'a}ri and T. Spasokukotskaja and F. Antoni and M. Staub",
year = "1985",
language = "English",
volume = "20",
pages = "173--182",
journal = "Ideggyogyaszati Szemle",
issn = "0019-1442",
publisher = "Ifjusagi Lap-es Konyvkiado Vallalat",
number = "3-4",

}

TY - JOUR

T1 - Pyrimidine salvage enzymes in human tonsil lymphocytes

T2 - II. Purification and properties of deoxycytidine kinase.

AU - Szyfter, K.

AU - Sasvári, M.

AU - Spasokukotskaja, T.

AU - Antoni, F.

AU - Staub, M.

PY - 1985

Y1 - 1985

N2 - Human tonsillar lymphocytes incorporate about 7-10 times more [3H]-deoxy-thymidine than [3H]-deoxycytidine at the same extracellular nucleoside concentration, and both incorporations could be inhibited by 10(-5) M arabinosyl-cytosine to 99%. On the other hand, the crude extract of these cells had about 10 times more deoxycytidine kinase than deoxythymidine kinase specific activity, as it was reported previously as well. Therefore, the differences in the labelling of these cells by [3H]-deoxycytidine and [3H]-thymidine cannot be simple due to the different levels of phosphorylating enzymes in the cells. The deoxycytidine kinase of human tonsillar lymphocytes was purified 38.6-fold by DEAE-Sephadex chromatography, and some kinetic and regulatory properties of the purified enzyme were investigated. Deoxycytidine kinase was eluted as a single peak from DEAE-Sephadex column and also from Sephadex G-100 column indicating a molecular weight of about 60 000; no isoenzymes could be detected. During the purification procedure an increase of the enzyme activity was observed suggesting the inhibition of the enzyme in the cells. An apparent Km of 13 microM for deoxycytidine and Ki of 55 microM for arabinosyl-cytosine were found for the tonsillar deoxycytidine kinase. The enzyme was strongly inhibited by dCTP, but not by the other deoxyribonucleoside triphosphates.

AB - Human tonsillar lymphocytes incorporate about 7-10 times more [3H]-deoxy-thymidine than [3H]-deoxycytidine at the same extracellular nucleoside concentration, and both incorporations could be inhibited by 10(-5) M arabinosyl-cytosine to 99%. On the other hand, the crude extract of these cells had about 10 times more deoxycytidine kinase than deoxythymidine kinase specific activity, as it was reported previously as well. Therefore, the differences in the labelling of these cells by [3H]-deoxycytidine and [3H]-thymidine cannot be simple due to the different levels of phosphorylating enzymes in the cells. The deoxycytidine kinase of human tonsillar lymphocytes was purified 38.6-fold by DEAE-Sephadex chromatography, and some kinetic and regulatory properties of the purified enzyme were investigated. Deoxycytidine kinase was eluted as a single peak from DEAE-Sephadex column and also from Sephadex G-100 column indicating a molecular weight of about 60 000; no isoenzymes could be detected. During the purification procedure an increase of the enzyme activity was observed suggesting the inhibition of the enzyme in the cells. An apparent Km of 13 microM for deoxycytidine and Ki of 55 microM for arabinosyl-cytosine were found for the tonsillar deoxycytidine kinase. The enzyme was strongly inhibited by dCTP, but not by the other deoxyribonucleoside triphosphates.

UR - http://www.scopus.com/inward/record.url?scp=0022177320&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022177320&partnerID=8YFLogxK

M3 - Article

VL - 20

SP - 173

EP - 182

JO - Ideggyogyaszati Szemle

JF - Ideggyogyaszati Szemle

SN - 0019-1442

IS - 3-4

ER -