1zmx
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(New page: 200px<br /><applet load="1zmx" size="450" color="white" frame="true" align="right" spinBox="true" caption="1zmx, resolution 3.10Å" /> '''Crystal structure of...)
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Revision as of 05:25, 21 November 2007
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Crystal structure of D. melanogaster deoxyribonucleoside kinase N64D mutant in complex with thymidine
Overview
The Drosophila melanogaster deoxyribonucleoside kinase (Dm-dNK) double, mutant N45D/N64D was identified during a previous directed evolution, study. This mutant enzyme had a decreased activity towards the natural, substrates and decreased feedback inhibition with dTTP, whereas the, activity with 3'-modified nucleoside analogs like 3'-azidothymidine (AZT), was nearly unchanged. Here, we identify the mutation N64D as being, responsible for these changes. Furthermore, we crystallized the mutant, enzyme in the presence of one of its substrates, thymidine, and the, feedback inhibitor, dTTP. The introduction of the charged Asp residue, appears to destabilize the LID region (residues 167-176) of the enzyme by, electrostatic repulsion and no hydrogen bond to the 3'-OH is made in the, substrate complex by Glu172 of the LID region. This provides a binding, space for more bulky 3'-substituents like the azido group in AZT but, influences negatively the interactions between Dm-dNK, substrates and, feedback inhibitors based on deoxyribose. The detailed picture of the, structure-function relationship provides an improved background for future, development of novel mutant suicide genes for Dm-dNK-mediated gene, therapy.
About this Structure
1ZMX is a Single protein structure of sequence from Drosophila melanogaster with SO4 and THM as ligands. Active as Deoxynucleoside kinase, with EC number 2.7.1.145 Full crystallographic information is available from OCA.
Reference
Structural basis for the changed substrate specificity of Drosophila melanogaster deoxyribonucleoside kinase mutant N64D., Welin M, Skovgaard T, Knecht W, Zhu C, Berenstein D, Munch-Petersen B, Piskur J, Eklund H, FEBS J. 2005 Jul;272(14):3733-42. PMID:16008571
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