1nn1
Crystal structure of human thymidylate kinase with ddTMP and AppNHp
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OverviewOverview
Nucleoside analogue prodrugs are dependent on efficient intracellular, stepwise phosphorylation to their triphosphate form to become, therapeutically active. In many cases it is this activation pathway that, largely determines the efficacy of the drug. To gain further understanding, of the determinants for efficient conversion by the enzyme thymidylate, kinase (TMPK) of clinically important thymidine monophosphate analogues to, the corresponding diphosphates, we solved the crystal structures of the, enzyme, with either ADP or the ATP analogue AppNHp at the phosphoryl donor, site, in complex with TMP, AZTMP (previous work), NH2TMP, d4TMP, ddTMP, and FLTMP (this work) at the phosphoryl acceptor site. In conjunction with, steady-state kinetic data, our structures shed light on the effect of, 3'-substitutions in the nucleoside monophosphate (NMP) sugar moiety on the, catalytic rate. We observe a direct correlation between the rate of, phosphorylation of an NMP and its ability to induce a closing of the, enzyme's phosphate-binding loop (P-loop). Our results show the drastic, effects that slight modifications of the substrates exert on the enzyme's, conformation and, hence, activity and suggest the type of substitutions, that are compatible with efficient phosphorylation by TMPK.
About this StructureAbout this Structure
1NN1 is a Single protein structure of sequence from Homo sapiens with MG, 2DT and ANP as ligands. Active as dTMP kinase, with EC number 2.7.4.9 Full crystallographic information is available from OCA.
ReferenceReference
Structures of human thymidylate kinase in complex with prodrugs: implications for the structure-based design of novel compounds., Ostermann N, Segura-Pena D, Meier C, Veit T, Monnerjahn C, Konrad M, Lavie A, Biochemistry. 2003 Mar 11;42(9):2568-77. PMID:12614151
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