Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.1.13.2 extracted from

  • Hang, J.Q.; Rajendran, S.; Yang, Y.; Li, Y.; In, P.W.K.; Overton, H.; Parkes, K.E.B.; Cammack, N.; Martin, J.A.; Klumpp, K.
    Activity of the isolated HIV RNase H domain and specific inhibition by N-hydroxyimides (2004), Biochem. Biophys. Res. Commun., 317, 321-329.
    View publication on PubMed
Search for more literature for 3.1.13.2

Cloned(Commentary)

Cloned (Comment) Organism
gene HXB2, expression as maltose-binding-protein fusion protein containing a TEV cleavae site in Escherichia coli strain BL21(DE3), functional expression of His-tagged isolated RNase H domain in Escherichia coli strain BL21(DE3) Human Immunodeficiency Virus

Protein Variants

Protein Variants Comment Organism
E478Q site-directed mutagenesis, RNase H domain active site mutation, inactive mutant Human Immunodeficiency Virus
additional information deletion of the 3 C-terminal residues of RNase H domain leads to strong inhibition of RNA cleavage activity Human Immunodeficiency Virus

Inhibitors

Inhibitors Comment Organism Structure
2-hydroxy-4H-isoquinoline-1,3-dione specific inhibition of isolated RNase H domain and of the full length reverse transcriptase, IC50 value for the isolated RNase H domain is 0.00043 mM Human Immunodeficiency Virus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
 Human Immunodeficiency Virus
Mn2+
-
 Human Immunodeficiency Virus

Organism

Organism UniProt Comment Textmining
Human Immunodeficiency Virus
-
 gene HXB2
-

Purification (Commentary)

Purification (Comment) Organism
recombinant maltose-binding-protein fusion protein from Escherichia coli strain BL21(DE3), the fused protein is cleaved off, purification by amylose affinity, and several steps of ion exchange chromatography and gel filtration, to over 95% purity, His-tagged isolated RNase H domain from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Human Immunodeficiency Virus

Reaction

Reaction Comment Organism Reaction ID
3'-end directed exonucleolytic cleavage of viral RNA-DNA hybrid RNase H domain, part of reverse transcriptase: active site residue E478 is essential for activity Human Immunodeficiency Virus
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
DNA-RNA hybrid + H2O formed by commercial R1 RNA and D1 DNA oligonucleotides, the His-tag affects the cleavage pattern of recombinant RNaseh domain Human Immunodeficiency Virus ?
-
 ?
additional information the isolated RNase H domain, part of reverse transcriptase, is catalytically active Human Immunodeficiency Virus ?
-
 ?

Subunits

Subunits Comment Organism
More the isolated RNase H domain is catalytically active with a similar activity as the full length reverse transcriptase Human Immunodeficiency Virus

Synonyms

Synonyms Comment Organism
HIV RNase H
-
 Human Immunodeficiency Virus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Human Immunodeficiency Virus

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.00043
-
 specific inhibition of isolated RNase H domain and of the full length reverse transcriptase, IC50 value for the isolated RNase H domain is 0.00043 mM Human Immunodeficiency Virus 2-hydroxy-4H-isoquinoline-1,3-dione