Substrate/Inhibitor Properties of Human Deoxycytidine Kinase (dCK) and Thymidine Kinases (Tk1 And Tk2) Towards the Sugar Moiety of Nucleosides,Including O′-Alkyl Analogues |
| |
| Authors: | Borys Kierdaszuk Krzysztof Krawiec Zygmunt Kazimierczuk Ulla Jacobsson Nils G Johansson Birgitte Munch-petersen |
| |
| Institution: | 1. University of Warsaw, Institute of Experimental Physics, Department of Biophysics , 93 Zwirki i Wigury St., PL-02089, Warsaw, Poland;2. University of Warsaw, Institute of Experimental Physics, Department of Biophysics , 93 Zwirki i Wigury St., PL-02089, Warsaw, Poland;3. Agricultural University, Institute of Chemistry , 26/30 Rakowiecka St., PL-02528, Warsaw;4. Department of Organic Chemistry , Royal Institute of Technology , S-100 44, Stockholm, Sweden;5. Medivir AB , Lunastigen 7, S-14144, Huddinge, Sweden;6. University of Roskilde, Department of Life Sciences and Chemistry , DK-4000, Roskilde, Denmark |
| |
| Abstract: | Abstract Nucleoside analogues with modified sugar moieties have been examined for their substrate/inhibitor specificities towards highly purified deoxycytidine kinase (dCK) and thymidine kinases (tetrameric high-affinity form of TK1, and TK2) from human leukemic spleen. In particular, the analogues included the mono-and di-O′-methyl derivatives of dC, dU and dA, syntheses of which are described. In general, purine nucleosides with modified sugar rings were feebler substrates than the corresponding cytosine analogues. Sugar-modified analogues of dU were also relatively poor substrates of TK1 and TK2, but were reasonably good inhibitors, with generally lower Ki values vs TK2 than TK1. An excellent discriminator between TK1 and TK2 was 3′-hexanoylamino-2′,3′-dideoxythymidine, with a Ki of ～600 μM for TK1 and ～0.1 μM for TK2. 3′-OMe-dC was a superior inhibitor of dCK to its 5′-O-methyl congener, consistent with possible participation of the oxygen of the (3′)-OH or (3′)-OMe as proton acceptor in hydrogen bonding with the enzyme. Surprisingly α-dT was a good substrate of both TK1 and TK2, with Ki values of 120 and 30 μM for TK1 and TK2, respectively; and a 3′-branched α-L-deoxycytidine analogue proved to be as good a substrate as its α-D-counterpart. Several 5 ′-substituted analogues of dC were |
| |
| Keywords: | |
|
|
