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 | 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 | 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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Human Immunodeficiency Virus | |
Mn2+ | - |
Human Immunodeficiency Virus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Human Immunodeficiency Virus | - |
gene HXB2 | - |
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 | 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 |
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 | 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 | Comment | Organism |
---|---|---|
HIV RNase H | - |
Human Immunodeficiency Virus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Human Immunodeficiency Virus |
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 |