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

BRENDA support

Literature summary extracted from

  • Chapados, B.R.; Chai, Q.; Hosfield, D.J.; Qiu, J.; Shen, B.; Tainer, J.A.
    Structural biochemistry of a type 2 RNase H: RNA primer recognition and removal during DNA replication (2001), J. Mol. Biol., 307, 541-556.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.26.4 type 2 enzyme, overexpression of wild-type in Escherichia coli BL21(DE3), and of selenomethionine mutant in Escherichia coli strain B834, a methionine-auxotroph BL21(DE3) derivative Archaeoglobus fulgidus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.26.4 crystallization of 40 mg/ml purified recombinant wild-type type 2 enzyme, and of recombinant selenomethionine enzyme mutant, with or without bound metal ions, hanging drop vapour diffusion method, mixing with equal volume of precipitant solution containing 10% PEG monomethylether 5000, 10% butanol, 20 mM sodium citrate, 100 mM Tris, pH 8.5, 3 days, formation of a cobalt-hexammine chloride complex, X-ray diffraction structure determination and analysis at 1.95-2.7 A resolution Archaeoglobus fulgidus

Protein Variants

EC Number Protein Variants Comment Organism
3.1.26.4 K143A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Archaeoglobus fulgidus
3.1.26.4 R146A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Archaeoglobus fulgidus
3.1.26.4 R46A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Archaeoglobus fulgidus
3.1.26.4 Y164A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Archaeoglobus fulgidus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.26.4 0.00006
-
 RNA-DNA*DNA hybrid recombinant wild-type enzyme, pH 8.0, 30°C Archaeoglobus fulgidus
3.1.26.4 0.0016
-
 RNA-DNA*DNA hybrid recombinant mutant R146A, pH 8.0, 30°C Archaeoglobus fulgidus
3.1.26.4 0.00188
-
 RNA-DNA*DNA hybrid recombinant mutant K143A, pH 8.0, 30°C Archaeoglobus fulgidus
3.1.26.4 0.00268
-
 RNA-DNA*DNA hybrid recombinant mutant Y164A, pH 8.0, 30°C Archaeoglobus fulgidus
3.1.26.4 0.0036
-
 RNA-DNA*DNA hybrid recombinant mutant R46A, pH 8.0, 30°C Archaeoglobus fulgidus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.26.4 Mg2+ dependent on Archaeoglobus fulgidus
3.1.26.4 additional information metal coordination in the active site Archaeoglobus fulgidus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.26.4 RNA-DNA*DNA hybrid + H2O Archaeoglobus fulgidus RNA primer recognition and removal during DNA replication DNA*DNA + 5'-phosphomononucleotides
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.1.26.4 Archaeoglobus fulgidus O29634 type 2 enzyme
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.26.4 recombinant enzymes from Escherichia coli Archaeoglobus fulgidus

Reaction

EC Number Reaction Comment Organism Reaction ID
3.1.26.4 Endonucleolytic cleavage to a 5'-phosphomonoester active site structure, substrate binding groove, molecular mechanism for specific RNA*DNA hybrid substrate recognition, binding, and cleavage, a general nuclease activity is necessary for catalysis Archaeoglobus fulgidus
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.26.4 additional information cleavage specificity Archaeoglobus fulgidus ?
-
 ?
3.1.26.4 RNA-DNA single-stranded chimera + H2O
-
 Archaeoglobus fulgidus ?
-
 ?
3.1.26.4 RNA-DNA*DNA hybrid + H2O RNA primer recognition and removal during DNA replication Archaeoglobus fulgidus DNA*DNA + 5'-phosphomononucleotides
-
 ?
3.1.26.4 RNA-DNA*DNA hybrid + H2O 7 M urea-denatured 18mer, no activity with untreated hybrid, specific removal of the RNA portion of the duplex Archaeoglobus fulgidus 8mer oligonucleotide
-
 ?
3.1.26.4 RNA-DNA*DNA hybrid + H2O 7 M urea-denatured 37mer, no activity with untreated hybrid, specific removal of the RNA portion of the duplex Archaeoglobus fulgidus oligonucleotides a 20mer, a 10mer, a 9mer, and a 7mer ?

Subunits

EC Number Subunits Comment Organism
3.1.26.4 More two-domain architecture Archaeoglobus fulgidus

Synonyms

EC Number Synonyms Comment Organism
3.1.26.4 More the enzyme belongs to the RNase H nuclease family Archaeoglobus fulgidus
3.1.26.4 ribonuclease H
-
 Archaeoglobus fulgidus
3.1.26.4 RNase H
-
 Archaeoglobus fulgidus
3.1.26.4 RNase HII
-
 Archaeoglobus fulgidus
3.1.26.4 type 2 RNase H
-
 Archaeoglobus fulgidus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.26.4 30
-
 assay at Archaeoglobus fulgidus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.26.4 0.133
-
 RNA-DNA*DNA hybrid recombinant wild-type enzyme, pH 8.0, 30°C Archaeoglobus fulgidus
3.1.26.4 0.192
-
 RNA-DNA*DNA hybrid recombinant mutant R146A, pH 8.0, 30°C Archaeoglobus fulgidus
3.1.26.4 0.2
-
 RNA-DNA*DNA hybrid recombinant mutant R46A and K143A, pH 8.0, 30°C Archaeoglobus fulgidus
3.1.26.4 0.208
-
 RNA-DNA*DNA hybrid recombinant mutant Y164A, pH 8.0, 30°C Archaeoglobus fulgidus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.26.4 8
-
 assay at Archaeoglobus fulgidus