Literature summary for 1.17.4.1 extracted from

Gurmu, D.; Dahlroth, S.L.; Haas, J.; Nordlund, P.; Erlandsen, H.
Expression, purification, crystallization and preliminary X-ray analysis of ORF60, the small subunit (R2) of ribonucleotide reductase from Kaposis sarcoma-associated herpesvirus (KSHV) (2010), Acta Crystallogr. Sect. F, 66, 734-737.

Cloned(Commentary)

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 (Commentary)

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

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

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 [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

Organism	UniProt	Comment	Textmining
Human gammaherpesvirus 8	-	an oncovirus belonging to the gamma-subfamily of human herpesviruses	-

Purification (Commentary)

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 and Products (Substrate)

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

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

Synonyms	Comment	Organism
class I RNR	-	Human gammaherpesvirus 8
RNR	-	Human gammaherpesvirus 8

General Information

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