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Literature summary for 3.1.26.13 extracted from

  • Farias, R.V.; Vargas, D.A.; Castillo, A.E.; Valenzuela, B.; Cote, M.L.; Roth, M.J.; Leon, O.
    Expression of an Mg2+-dependent HIV-1 RNase H construct for drug screening (2011), Antimicrob. Agents Chemother., 55, 4735-4741.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.26.13

Cloned(Commentary)

Cloned (Comment) Organism
expression of RNase H, in an N-terminally His-tagged construct designed to encode the p51 subunit joined by a linker to the thumb (T), connection (C), and RNase H (R) domains of p66, in Escherichia coli strain MIC2067(DE3) lacking endogenous RNase HI and HII as soluble protein. The construct G provides sufficient RNase H activity to complement Escherichia coli growth at 42°C Human immunodeficiency virus 1

Protein Variants

Protein Variants Comment Organism
E706Q site-directed mutagenesis of HIV-1 reverse transcriptase, inactive mutant Human immunodeficiency virus 1
E706Q site-directed mutagenesis of the recombinant mutant construct G, E706 in construct G corresponds to E478 in HIV-1 reverse transcriptase, inactive mutant Human immunodeficiency virus 1
additional information construction of an N-terminally His-tagged mutant p51-G-TCR construct designed to encode the p51 subunit joined by a linker to the thumb (T), connection (C), and RNase H (R) domains of p66, the p51-G-TCR RNase H construct displays Mg2+-dependent activity using a fluorescent nonspecific assay and shows the same cleavage pattern as HIV-1 reverse transcriptase on substrates that mimic the tRNA removal required for second-strand transfer reactions. The RNase H of the p51-G-TCR RNase H construct and wild-type HIV-1 reverse transcriptase have similar Kms for an RNA-DNA hybrid substrate and show similar inhibition kinetics to two known inhibitors of the HIV-1 reverse transcriptase RNase H, molecular modeling Human immunodeficiency virus 1

Inhibitors

Inhibitors Comment Organism Structure
ARK-2452
-
 Human immunodeficiency virus 1
beta-thujaplicinol
-
 Human immunodeficiency virus 1

Metals/Ions

Metals/Ions Comment Organism Structure
K+ required Human immunodeficiency virus 1
Mg2+ dependent on Human immunodeficiency virus 1

Organism

Organism UniProt Comment Textmining
Human immunodeficiency virus 1
-
 HIV-1
-

Purification (Commentary)

Purification (Comment) Organism
soluble recombinant RNase H, in an N-terminally His-tagged construct, p51-G-TCR construct, designed to encode the p51 subunit joined by a linker to the thumb (T), connection (C), and RNase H (R) domains of p66, from Escherichia coli strain MIC2067(DE3) by nickel affinity chromatography Human immunodeficiency virus 1

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information RNA-DNA substrate in a tRNA removal assay, the cleavage patterns for the recombinant HIV-1 reverse transcriptase and mutant p51-G-TCR construct correspond to the release of the 11mer RNA, corresponding with the authentic cleavage identified in vivo. The relative activity of the isolated HIV-1 RNaseH(p51-G-TCR) is comparable to that of the full-length HIV reverse transcriptase Human immunodeficiency virus 1 ?
-
 ?

Synonyms

Synonyms Comment Organism
RNase H
-
 Human immunodeficiency virus 1

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Human immunodeficiency virus 1

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Human immunodeficiency virus 1

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.000176
-
 pH 8.0, 37°C, purified recombinant mutant p51-G-TCR construct Human immunodeficiency virus 1 beta-thujaplicinol
0.000186
-
 pH 8.0, 37°C, purified recombinant mutant p51-G-TCR construct Human immunodeficiency virus 1 ARK-2452
0.000257
-
 pH 8.0, 37°C, purified recombinant wild-type HIV-1 reverse transcriptase RNase H Human immunodeficiency virus 1 beta-thujaplicinol
0.00047
-
 pH 8.0, 37°C, purified recombinant wild-type HIV-1 reverse transcriptase RNase H Human immunodeficiency virus 1 ARK-2452