However, this is not the case with dCTP, which appeared to act as a competitive in- hibitor of the phosphate donor (ATP) and as a noncompetitive inhibitor of the phosphate acceptor (deoxycytidine or cytosine arabinoside). These results could be explained if one assumes that there is a second site of inhibition of dCTP. Although the inhibition produced by dCTP appeared to be primarily competitive with the phosphate donor, it is possible that the inhibition may have been also weakly competitive with the phosphate ac- ceptor, cytosine arabinoside being more sensitive to this in- hibition than deoxycytidine. The similarity in structure of deoxycytidine and dCTP would favor this proposal. This type of inhibition has been observed by Okazaki and Kornberg (18), who reported that dTTP appeared to act as a competitive in- hibitor of the phosphate acceptor (deoxythymidine) of deoxy- thymidine kinase of Escherkhiu coli.
In kinetic studies with deoxycytidine kinase, it was found that both deoxycytidine and cytosine arabinoside were competitive inhibitors of each other. However, deoxycytidine was a much more potent competitive inhibitor than cytosine arabinodide. This result is probably due to the greater afhnity of deoxycyti- dine for the phosphate acceptor site than cytosine arabinoside and to the phosphorylation and release of cytosine arabinoside from the acceptor site, thus making this site more accessible to deoxycytidine.
The data presented in this paper suggest that the reversal by deoxycytidine of the inhibitory effects produced by cytosine arabinoside on mammalian cells (3, 7) must involve to some ex- tent interactions with deoxycytidine kinase. The competitive inhibition of the phosphorylation of cytosine arabinoside by deoxycytidine and the inhibition of deoxycytidine kinase by cytosine deoxynucleotides (8, 9) would result in a decreased con- version of cytosine arabinoside to its active form. The increased sensitivity of cytosine arabinoside to the inhibitory effects of cytosine deoxynucleotides suggests that tumor cells with a larger intracellular pool of these deoxynucleotides should be more resistant to cytosine arabinoside.
Acknowledgments-We wish to thank Mrs. Joyce Cramer and Mrs. Barbara Morcaldi for their excellent technical assistance
However, this is not the case with dCTP, which appeared to act as a competitive in- hibitor of the phosphate donor (ATP) and as a noncompetitive inhibitor of the phosphate acceptor (deoxycytidine or cytosine arabinoside). These results could be explained if one assumes that there is a second site of inhibition of dCTP. Although the inhibition produced by dCTP appeared to be primarily competitive with the phosphate donor, it is possible that the inhibition may have been also weakly competitive with the phosphate ac- ceptor, cytosine arabinoside being more sensitive to this in- hibition than deoxycytidine. The similarity in structure of deoxycytidine and dCTP would favor this proposal. This type of inhibition has been observed by Okazaki and Kornberg (18), who reported that dTTP appeared to act as a competitive in- hibitor of the phosphate acceptor (deoxythymidine) of deoxy- thymidine kinase of Escherkhiu coli.In kinetic studies with deoxycytidine kinase, it was found that both deoxycytidine and cytosine arabinoside were competitive inhibitors of each other. However, deoxycytidine was a much more potent competitive inhibitor than cytosine arabinodide. This result is probably due to the greater afhnity of deoxycyti- dine for the phosphate acceptor site than cytosine arabinoside and to the phosphorylation and release of cytosine arabinoside from the acceptor site, thus making this site more accessible to deoxycytidine.The data presented in this paper suggest that the reversal by deoxycytidine of the inhibitory effects produced by cytosine arabinoside on mammalian cells (3, 7) must involve to some ex- tent interactions with deoxycytidine kinase. The competitive inhibition of the phosphorylation of cytosine arabinoside by deoxycytidine and the inhibition of deoxycytidine kinase by cytosine deoxynucleotides (8, 9) would result in a decreased con- version of cytosine arabinoside to its active form. The increased sensitivity of cytosine arabinoside to the inhibitory effects of cytosine deoxynucleotides suggests that tumor cells with a larger intracellular pool of these deoxynucleotides should be more resistant to cytosine arabinoside.Acknowledgments-We wish to thank Mrs. Joyce Cramer and Mrs. Barbara Morcaldi for their excellent technical assistance
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