Cloned (Comment) | Organism |
---|---|
recombinant overexpression of the ORF60 protein, subunit R2, with an N-terminal extension, MGPHHHHHHLESTSLYKKAGS in Escherichia coli BL21 (DE3) cells. The N-terminal extension encompasses an attB1 site for the gateway recombination event and a His tag to facilitate protein purification | Human gammaherpesvirus 8 |
Crystallization (Comment) | Organism |
---|---|
small subunit R2 of ribonucleotide reductase, at 4°C and 20°C, 10 mg/ml protein in 20 mM HEPES, pH 7.5, 300 mM NaCl, 10% glycerol, 0.1 mg/ml chymotrypsin, and 0.1 M hexamine cobalt(III) chloride, is mixed with optimized reservoir solution containing 14% w/v PEG 8000, 0.2 M MgCl2,0.1 M Tris-HCl pH 8.2, optimization of the crystallization conditions, X-ray diffraction structure determination and analysis at 2.0 A resolution | Human gammaherpesvirus 8 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | Glu64 is found in the viral protein in the position that is usually occupied by a metal-coordinating aspartate in other R2s | Human gammaherpesvirus 8 |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
31000 | - |
4 * 37 442.98, subunit R2, mass spectrometry, 4 * 31000, recombinant subunit R2, SDS-PAGE | Human gammaherpesvirus 8 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Human gammaherpesvirus 8 | - |
an oncovirus belonging to the gamma-subfamily of human herpesviruses | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged ORF60 protein, subunit R2, from Escherichia coli BL21 (DE3) by metal affinity chromatography and gel filtration | Human gammaherpesvirus 8 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme catalyzes the reduction of all four ribonucleotides to their corresponding deoxyribonucleotides, the R2 subunit contains a di-iron site, which generates a free radical by the reductive cleavage of molecular oxygen. The free radical is subsequently transferred to the R1 subunit, activating the nucleotide substrate for catalysis | Human gammaherpesvirus 8 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | class I RNRs are composed of a heterotetramer, which is in turn composed of two homodimers of the R1 and R2 subunits. The R1 subunit contains the active site as well as the sites for allosteric regulation | Human gammaherpesvirus 8 |
tetramer | 4 * 37 442.98, subunit R2, mass spectrometry, 4 * 31000, recombinant subunit R2, SDS-PAGE | Human gammaherpesvirus 8 |
Synonyms | Comment | Organism |
---|---|---|
class I RNR | - |
Human gammaherpesvirus 8 |
RNR | - |
Human gammaherpesvirus 8 |
General Information | Comment | Organism |
---|---|---|
physiological function | the allosterically regulated enzyme is responsible for the reduction of all four ribonucleotides to their corresponding deoxyribonucleotides, dNTPs, which are the building blocks of DNA. This reaction involves a free radical, the activity of RNR regulates the cellular levels of the dNTP pool, ensuring that precise DNA replication and repair occurs | Human gammaherpesvirus 8 |