Phosphorylation of dGMP analogs by vaccinia virus TMP kinase and human GMP kinase.
Vaccinia virus thymidylate kinase, although similar in sequence to human TMP kinase, has broader substrate specificity and phosphorylates (E)-5-(2-bromovinyl)-dUMP and dGMP. Modified guanines such as glyoxal-dG, 8-oxo-dG, O(6)-methyl-dG, N(2)-ethyl-dG and N(7)-methyl-dG were found present in cancer cell DNA. Alkylated and oxidized dGMP analogs were examined as potential substrates for vaccinia TMP kinase and also for human TMP and GMP kinases. Molecular models obtained from structure-based docking rationalized the enzymatic data. All tested nucleotides are found surprisingly substrates of vaccinia TMP kinase and also of human GMP kinase. Interestingly, O(6)-methyl-dGMP is the only analog specific for the vaccinia enzyme. Thus, O(6)-Me-dGMP could be useful for designing new compounds of medical interest either in antipoxvirus therapy or in experimental combined gene/chemotherapy of cancer. These results also provide new insights regarding dGMP analog reaction with human GMP kinase and their slow recycling by salvage pathway nucleotide kinases.
Auvynet
Constance
C
Université Pierre et Marie Curie, UR, Paris, France.
Topalis
Dimitri
D
Caillat
Christophe
C
Munier-Lehmann
Hélène
H
Seclaman
Edward
E
Balzarini
Jan
J
Agrofoglio
Luigi André
LA
Kaminski
Pierre Alexandre
PA
Meyer
Philippe
P
Deville-Bonne
Dominique
D
El Amri
Chahrazade
C
D000998:Antiviral Agents / Q000737:chemistry; D003848:Deoxyguanine Nucleotides / Q000378:metabolism*; D015195:Drug Design; D051528:Guanylate Kinases / Q000378:metabolism*; D006801:Humans; D009703:Nucleoside-Phosphate Kinase / Q000378:metabolism*; D010766:Phosphorylation; D013379:Substrate Specificity; D014616:Vaccinia virus / Q000201:enzymology*
D000998:Antiviral Agents; D003848:Deoxyguanine Nucleotides; C007257:2'-deoxyguanosine 5'-phosphate; D009703:Nucleoside-Phosphate Kinase; D051528:Guanylate Kinases; C020093:dTMP kinase