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PMID: 2480079 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Phosphorylation of 3'-azido-2',3'-dideoxyuridine and preferential inhibition of human and simian immunodeficiency virus reverse transcriptases by its 5'-triphosphate.

Antimicrobial agents and chemotherapy ·Vol. 33 ·No. 10 ·1989-10-00 ·Pages 1729-34

Eriksson BF, Chu CK, Schinazi RF

Abstract

3'-Azido-2',3'-dideoxyuridine-5'-triphosphate was found to be a potent and highly selective inhibitor of human immunodeficiency virus type 1 and simian immunodeficiency virus reverse transcriptases. The affinity of 3'-azido-2',3'-dideoxyuridine-5'-triphosphate for the reverse transcriptases was similar to that observed for 3'-azido-3'-deoxythymidine-5'-triphosphate. Both compounds were competitive inhibitors with respect to the normal substrate dTTP and served at least 100 times better as substrates than did dTTP. In contrast, cellular DNA polymerase alpha showed an about 60-times-higher preference for dTTP as substrate than for either inhibitor. The phosphorylation of thymidine in human peripheral blood mononuclear cell extracts was inhibited in a competitive manner by both 3'-azido-2',3'-dideoxyuridine and 3'-azido-3'-deoxythymidine, with apparent inhibition constants of 290 and 3.4 microM, respectively. The Michaelis-Menten constant, Km, for thymidine was 7.0 microM. 3'-Azido-2',3'-dideoxyuridine and 3'-azido-3'-deoxythymidine both served as substrates, with apparent Km values of 67 and 1.4 microM, respectively. The maximal rates of phosphorylation with 3'-azido-2',3'-dideoxyuridine and 3'-azido-3'-deoxythymidine were 40 and 30%, respectively, of the rate with thymidine. The different affinities of 3'-azido-2',3'-dideoxyuridine and 3'-azido-3'-deoxythymidine for the thymidine kinase and the Km values observed with these compounds as substrates may explain the difference in effects on human immunodeficiency virus type 1 replication in infected peripheral blood mononuclear cells observed when equimolar concentrations of the two compounds are compared.

MeSH Terms
Cells, Cultured Chromatography, High Pressure Liquid DNA Polymerase II/antagonists & inhibitors HIV/drug effects,enzymology Humans Kinetics Phosphorylation Phosphotransferases/metabolism Reverse Transcriptase Inhibitors Simian Immunodeficiency Virus/drug effects,enzymology Thymidine Kinase/metabolism Zidovudine/analogs & derivatives,pharmacology
Chemicals
Reverse Transcriptase Inhibitors Zidovudine Phosphotransferases Thymidine Kinase nucleoside phosphotransferase DNA Polymerase II 3'-azido-2',3'-dideoxyuridine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Eriksson B F
Veterans Administration Medical Center (Atlanta), Decatur, Georgia 30033.
Chu C K
Schinazi R F
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Article Info
Journal
Antimicrobial agents and chemotherapy
Abbr.
Antimicrob Agents Chemother
ISSN
0066-4804
DOI
10.1128/AAC.33.10.1729
Published
1989-10-00
Pages
1729-34
Language
English
Region
United States
NLM ID
0315061
PMCID
PMC172746
Subset
IM
Grants
NIAID NIH HHS · AI 25899 · United States
NIAID NIH HHS · AI 26055 · United States
External Links
PubMed source DOI: 10.1128/AAC.33.10.1729 PMC Full Text View journal details
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