X-Ray Analysis of Azido-Thymidine Diphosphate Binding to Nucleoside Diphosphate Kinase

• Published in: Proceedings of the National Academy of Sciences - PNAS 1997-07, Vol.94 (14), p.7162-7165
• Main Authors: Xu, Yingwu, Sellam, Olivier, Morera, Solange, Sarfati, Simon, Biondi, Ricardo, Veron, Michel, Janin, Joel
• Format: Article
• Language: English
• Subjects: Active sites; Animals; Binding Sites; Biochemistry; Biological Sciences; Crystallography, X-Ray; Dictyostelium; Dictyostelium discoideum; Diphosphates; Diphosphates - chemistry; Diphosphates - metabolism; Enzyme substrates; Enzymes; Hydrogen bonds; Molecular Sequence Data; Mutation; Net domestic product; Nucleoside-Diphosphate Kinase - chemistry; Nucleoside-Diphosphate Kinase - metabolism; Nucleosides; Nucleotides; Phosphates; X-rays; Zidovudine - chemistry; Zidovudine - metabolism
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.94.14.7162

To be effective as antiviral agent, AZT (3′-azido-3′-deoxythymidine) must be converted to a triphosphate derivative by cellular kinases. The conversion is inefficient and, to understand why AZT diphosphate is a poor substrate of nucleoside diphosphate (NDP) kinase, we determined a 2.3- angstrom x-ray structure of a complex with the N119A point mutant of Dictyostelium NDP kinase. It shows that the analog binds at the same site and, except for the sugar ring pucker which is different, binds in the same way as the natural substrate thymidine diphosphate. However, the azido group that replaces the 3′OH of the deoxyribose in AZT displaces a lysine side chain involved in catalysis. Moreover, it is unable to make an internal hydrogen bond to the oxygen bridging the β - and γ -phosphate, which plays an important part in phosphate transfer.