Steady state kinetics and inhibition of HIV-1 reverse transcriptase by a non-nucleoside dipyridodiazepinone, BI-RG-587, using a heteropolymeric template
| Autor: | Peter M. Grob, Anthony Shrutkowski, John Miglietta, Mark T. Skoog, Elizabeth B. Kopp, Cheng-Kon Shih |
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| Jazyk: | angličtina |
| Rok vydání: | 1991 |
| Předmět: |
chemistry.chemical_classification
Base Sequence Stereochemistry Pyridines Molecular Sequence Data Processivity Templates Genetic Biology Molecular biology Reverse transcriptase A-site Kinetics chemistry Genetics HIV-1 RNA Viral Reverse Transcriptase Inhibitors Nucleotide Electrophoresis Polyacrylamide Gel Enzyme kinetics Steady state (chemistry) Nevirapine Binding site Nucleoside |
| Popis: | Steady state kinetics and inhibition by a dipyridodiazepinone of the reverse transcriptase from human immunodeficiency virus type 1 (HIV) were studied using a heteropolymeric RNA template with a sequence from the authentic initiation site on the HIV genome. For addition of the first deoxynucleotide to primer, kcat/KM is 0.05 (nM-min)-1 and KM is 10 nM. When all 4 deoxynucleotide triphosphates are present and processive synthesis occurs, catalysis is less efficient; kcat/KM = .0077 (nM-min)-1 and KM = 100 nM for dATP. These results are consistent with a rate determining conformation change involved in translocation of the enzyme along the template. Inhibition by the dipyridodiazepinone BI-RG-587 is noncompetitive with respect to both nucleotide and template-primer; this compound decreases Vmax but does not affect KM. Thus, this inhibitor binds to a site distinct from the substrate binding sites with Ki of 220 nM. Inhibition by BI-RG-587 results in a uniform decrease in amount of products of all lengths rather than a shift from longer to shorter products, suggesting the inhibitor does not affect processivity of reverse transcriptase. |
| Databáze: | OpenAIRE |
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