Steady state kinetics and inhibition of HIV-1 reverse transcriptase by a non-nucleoside dipyridodiazepinone, BI-RG-587, using a heteropolymeric template.
Nucleic acids research (1991), Volume 19, Page 3035
Abstract:
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.
Polymerases:
Topics:
Kinetic Parameters
Status:
new | topics/pols set | partial results | complete | validated |
Results:
Polymerase | Reference | Property | Result | Context |
---|---|---|---|---|
HIV RT | Kopp EB1991 | KM | 10nM | Reaction: Nucleotide incorporation; Substrate: dCTP; Technique: Steady State |
HIV RT | Kopp EB1991 | KM | 40nM | Reaction: Nucleotide incorporation; Substrate: dTTP; Technique: Steady State |
HIV RT | Kopp EB1991 | KM | 20nM | Reaction: Nucleotide incorporation; Substrate: dGTP; Technique: Steady State |
HIV RT | Kopp EB1991 | KM | 100nM | Reaction: Nucleotide incorporation; Substrate: dATP; Technique: Steady State |
HIV RT | Kopp EB1991 | kcat | 0.5 /minute | Reaction: Nucleotide incorporation; Substrate: dCTP; Technique: Steady State |
HIV RT | Kopp EB1991 | kcat | 0.43 /minute | Reaction: Nucleotide incorporation; Substrate: dTTP; Technique: Steady State |
HIV RT | Kopp EB1991 | kcat | 0.12 /minute | Reaction: Nucleotide incorporation; Substrate: dGTP; Technique: Steady State |
HIV RT | Kopp EB1991 | kcat | 0.77 /minute | Reaction: Nucleotide incorporation; Substrate: dATP; Technique: Steady State |