X-ray analysis of azido-thymidine diphosphate binding to nucleoside diphosphate kinase.
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-A 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 beta- and gamma-phosphate, which plays an important part in phosphate transfer.
Xu
Y
Y
Laboratoire d'Enzymologie et de Biochimie Structurales, Unité Propre de Recherche 9063, Centre National de la Recherche Scientifique, 91198 Gif-sur-Yvette, France.
Sellam
O
O
Moréra
S
S
Sarfati
S
S
Biondi
R
R
Véron
M
M
Janin
J
J
D000818:Animals; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004023:Dictyostelium; D011756:Diphosphates / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry* / Q000378:metabolism; D015215:Zidovudine / Q000737:chemistry* / Q000378:metabolism
D011756:Diphosphates; D015215:Zidovudine; D009701:Nucleoside-Diphosphate Kinase