Information on EC 1.17.4.1 - ribonucleoside-diphosphate reductase and Organism(s) Homo sapiens and UniProt Accession P31350

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UNIPROT: P31350


The taxonomic range for the selected organisms is: Homo sapiens

The enzyme appears in selected viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.17.4.1
-
RECOMMENDED NAME
GeneOntology No.
ribonucleoside-diphosphate reductase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2'-deoxyribonucleoside 5'-diphosphate + thioredoxin disulfide + H2O = ribonucleoside 5'-diphosphate + thioredoxin
show the reaction diagram
proposed mechanism
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
guanosine deoxyribonucleotides de novo biosynthesis II
-
-
superpathway of pyrimidine deoxyribonucleotides de novo biosynthesis (E. coli)
-
-
pyrimidine deoxyribonucleotides biosynthesis from CTP
-
-
pyrimidine deoxyribonucleotides de novo biosynthesis I
-
-
adenosine deoxyribonucleotides de novo biosynthesis II
-
-
adenosine deoxyribonucleotides de novo biosynthesis
-
-
guanosine deoxyribonucleotides de novo biosynthesis I
-
-
pyrimidine deoxyribonucleotides de novo biosynthesis IV
-
-
pyrimidine deoxyribonucleotides de novo biosynthesis III
-
-
purine metabolism
-
-
pyrimidine metabolism
-
-
Purine metabolism
-
-
Pyrimidine metabolism
-
-
Glutathione metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
2'-deoxyribonucleoside-5'-diphosphate:thioredoxin-disulfide 2'-oxidoreductase
This enzyme is responsible for the de novo conversion of ribonucleoside diphosphates into deoxyribonucleoside diphosphates, which are essential for DNA synthesis and repair. There are three types of this enzyme differing in their cofactors. Class Ia enzymes contain a diiron(III)-tyrosyl radical, class Ib enzymes contain a dimanganese-tyrosyl radical, and class II enzymes contain adenosylcobalamin. In all cases the cofactors are involved in generation of a transient thiyl (sulfanyl) radical on a cysteine residue, which attacks the substrate, forming a ribonucleotide 3'-radical, followed by water loss to form a ketyl (alpha-oxoalkyl) radical. The ketyl radical is reduced to 3'-keto-deoxynucleotide concomitant with formation of a disulfide anion radical between two cysteine residues. A proton-coupled electron-transfer from the disulfide radical to the substrate generates a 3'-deoxynucleotide radical, and the final product is formed when the hydrogen atom that was initially removed from the 3'-position of the nucleotide by the thiyl radical is returned to the same position. The disulfide bridge is reduced by the action of thioredoxin. cf. EC 1.1.98.6, ribonucleoside-triphosphate reductase (formate) and EC 1.17.4.2, ribonucleoside-triphosphate reductase (thioredoxin).
CAS REGISTRY NUMBER
COMMENTARY hide
9047-64-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
small subunit RIR2
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
a conserved cluster of charged residues, including Lys95, Glu98, Glu-05, and Glu174, at the interface may function as an ionic lock for small subunit M2 homodimer. The transfection of the wild-type small subunit M2 but not the K95E mutant rescues theG1/S phase cell cycle arrest and cell growth inhibition caused by the siRNA knockdown of M2
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
CDP + reduced thioredoxin
2'-dCDP + thioredoxin disulfide + H2O
show the reaction diagram
-
-
-
-
?
nucleoside 5'-diphosphate + glutaredoxin
2'-deoxynucleoside 5'-diphosphate + glutaredoxin disulfide + H2O
show the reaction diagram
-
class Ia RNRs
-
-
?
nucleoside 5'-diphosphate + thioredoxin
2'-deoxynucleoside 5'-diphosphate + thioredoxin disulfide + H2O
show the reaction diagram
-
class Ia RNRs
-
-
?
ribonucleoside 5'-diphosphate + thioredoxin
2'-deoxyribonuleoside 5'-diphosphate + thioredoxin disulfide + H2O
show the reaction diagram
ribonucleoside diphosphate + reduced thioredoxin
2'-deoxyribonucleoside diphosphate + oxidized thioredoxin + H2O
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
nucleoside 5'-diphosphate + glutaredoxin
2'-deoxynucleoside 5'-diphosphate + glutaredoxin disulfide + H2O
show the reaction diagram
-
class Ia RNRs
-
-
?
nucleoside 5'-diphosphate + thioredoxin
2'-deoxynucleoside 5'-diphosphate + thioredoxin disulfide + H2O
show the reaction diagram
-
class Ia RNRs
-
-
?
ribonucleoside 5'-diphosphate + thioredoxin
2'-deoxyribonuleoside 5'-diphosphate + thioredoxin disulfide + H2O
show the reaction diagram
-
the essential enzyme catalyzes the rate-limiting step in dNTP production for DNA synthesis
-
-
?
ribonucleoside diphosphate + reduced thioredoxin
2'-deoxyribonucleoside diphosphate + oxidized thioredoxin + H2O
show the reaction diagram
-
enzyme catalyzes the first unique step in DNA synthesis
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
diferric(III)-tyrosyl radical cofactor
-
class I enzymes
-
glutaredoxin
-
class Ia RNRs
thioredoxin
additional information
-
cofactor specificity and binding, role in reaction, overview
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
-
monomers A and B exhibit mono- and binuclear iron occupancy, the active site iron coordination environment, involving E131, H134, D100, E194, E228, and H231, is different between monomers A and B, binding structure, overview. Mobility and accessibility of the radical iron center, and radical transfer pathway, overview
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(-)-epicatechin
-
interacts with the R2 protein, leading to a loss of the tyrosyl radical EPR signal. Proliferation of cells exposed to (-)-epicatechin is downregulated, and deoxyribonucleotide levels are significantly diminished
(2E)-2-(anthracen-9-ylmethylidene)-N-hydroxyhydrazinecarboximidamide
-
i.e. ABNM-13, application leads to significant alterations of deoxyribonucleoside triphosphate pool balance and a highly significant decrease of incorporation of radiolabeled cytidine into DNA of HL-60 cells. Diminished ribonucleotide reductase activity causes replication stress which is consistent with activation of Chk1 and Chk2, resulting in downregulation/degradation of Cdc25A. Cdc25B is upregulated, leading to dephosphorylation and activation of Cdk1. The combined disregulation of Cdc25A and Cdc25B is the most likely cause for ABNM-13 induced S-phase arrest
1,10-phenanthroline
-
-
2,3,4-Trihydroxybenzamide
-
-
2,3,4-trihydroxybenzohydroxamic acid
-
0.0035 mM, 50% inhibition
2,3-dihydroxybenzohydroxamic acid
-
0.008 mM, 50% inhibition
2,4-dichlorobenzohydroxamic acid
-
0.45 mM, 50% inhibition
2,4-dihydroxybenzohydroxamic acid
-
0.3 mM, 50% inhibition
2,5-dihydroxybenzohydroxamic acid
-
0.2 mM, 50% inhibition
2,6-dihydroxybenzohydroxamic acid
-
0.1 mM, 50% inhibition
2-(diphenylmethylidene)-N,N-dimethylhydrazinecarbothioamide
-
metal chelator, significantly decreases ribonucleotide reductase activity, whereas the NADPH/NADP+ total ratio is not reduced
2-acetylpyridine N,N-dimethylthiosemicarbazonato-N,N,S-dichlorogallium(III)
-
-
2-acetylpyridine N-pyrrolidinylthiosemicarbazonato-N,N,S-dichlorogallium(III)
-
-
2-aminobenzohydroxamic acid
-
0.12 mM, 50% inhibition
2-furan-3-ylbenzaldehyde N-(4-hydroxyphenyl)thiosemicarbazone
-
-
2-furan-3-ylbenzaldehyde N-phenylthiosemicarbazone
-
-
2-hydroxy-3-methylbenzohydroxamic acid
-
0.15 mM, 50% inhibition
2-hydroxy-4-aminobenzohydroxamic acid
-
0.2 mM, 50% inhibition
2-hydroxybenzaldehyde N-(4-chlorophenyl)thiosemicarbazone
-
-
2-hydroxybenzaldehyde N-phenylthiosemicarbazone
-
-
2-hydroxybenzohydroxamic acid
-
0.15 mM, 50% inhibition
2-thiophen-2-ylbenzaldehyde N-(4-chlorophenyl)thiosemicarbazone
-
-
2-thiophen-2-ylbenzaldehyde N-phenylthiosemicarbazone
-
-
2-[di(pyridin-2-yl)methylidene]-N,N-dimethylhydrazinecarbothioamide
-
metal chelator, significantly decreases ribonucleotide reductase activity, whereas the NADPH/NADP+ total ratio is not reduced
3,4,5-Trihydroxybenzamide
-
-
3,4,5-Trihydroxybenzohydroxamic acid
-
0.01 mM, 50% inhibition
3,4,5-Trihydroxybenzoic acid
-
-
3,4,5-trimethoxybenzohydroxamic acid
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0.1 mM, 50% inhibition
3,4-diaminobenzohydroxamic acid
-
0.04 mM, 50% inhibition
3,4-dichlorobenzohydroxamic acid
-
0.3 mM, 50% inhibition
3,4-Dihydroxybenzamide
-
-
3,4-dihydroxybenzohydroxamic acid
-
0.03 mM, 50% inhibition
3,4-dimethoxybenzohydroxamic acid
-
0.3 mM, 50% inhibition
3,4-dimethylbenzohydroxamic acid
-
0.3 mM, 50% inhibition
3,5-diamino-1H-1,2,4-triazole
3,5-diaminopyridine-2-carboxaldehyde thiosemicarbazone
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-
3,5-dihydroxybenzohydroxamic acid
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0.4 mM, 50% inhibition
3-amino-4-methylpyridine-2-carboxaldehyde thiosemicarbazone
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-
3-aminobenzohydroxamic acid
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0.35 mM, 50% inhibition
3-aminopyridine-2-carboxaldehyde thiosemicarbazone
3-aminopyridine-2-carboxaldehyde-thiosemicarbazone
-
i.e. 3-AP, phase I study in combination with high dose cytarabine in patients with advanced myeloid leukemia, resulting in enhanced cytarabine cytotoxicity with possible methemoglobinemia, overview
3-hydroxybenzohydroxamic acid
-
0.35 mM, 50% inhibition
3-methyl aminopyridine-2-carboxaldehyde thiosemicarbazone
-
-
4-aminobenzohydroxamic acid
-
0.15 mM, 50% inhibition
4-dimethylaminobenzohydroxamic acid
-
0.5 mM, 50% inhibition
4-hydroxy-3-methoxybenzaldehyde N-(4-chlorophenyl)thiosemicarbazone
-
-
4-hydroxy-3-methoxybenzaldehyde N-phenylthiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(2-chlorophenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(2-hydroxyphenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(2-methoxyphenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(2-methylphenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(2-nitrophenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(3-chlorophenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(3-hydroxyphenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(3-methylphenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(4-chlorophenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(4-hydroxyphenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(4-methylphenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-(4-nitrophenyl)thiosemicarbazone
-
-
4-hydroxybenzaldehyde N-phenylthiosemicarbazone
-
-
4-hydroxybenzohydroxamic acid
-
0.30 mM, 50% inhibition
4-methoxybenzohydroxamic acid
-
0.5 mM, 50% inhibition
4-methylaminobenzohydroxamic acid
-
0.33 mM, 50% inhibition
4-nitrobenzohydroxamic acid
-
0.5 mM, 50% inhibition
5-(1-Aziridinyl)-2,4-dinitrobenzamide
-
-
5-amino-4-morpholinomethylpyridine-2-carboxaldehyde thiosemicarbazone
-
-
5-aminopyridine-2-carboxaldehyde thiosemicarbazone
-
-
5-hydroxy-4-methyl-1-formylisoquinoline thiosemicarbazone
-
-
5-methyl-4-amino-1-formylisoquinoline thiosemicarbazone
-
-
6-chloro-9H-(3-C-methyl-2,3-di-O-acetyl-5-O-benzoyl-beta-D-ribofuranosyl)purine
-
-
Acetohydroxamic acid
-
1 mM, 50% inhibition
benzohydroxamic acid
-
0.4 mM, 50% inhibition
caracemide
-
-
chlorambucil
-
-
cisplatin
-
-
clofarabine
-
an adenosine analogue is used in the treatment of refractory leukemias. Its mode of cytotoxicity is associated in part with the triphosphate functioning as an allosteric reversible inhibitor of hRNR, rapid inactivation
clofarabine diphosphate
-
ClFDP, a C-site slow-binding, reversible inhibitor, mechanism of inhibition via altering the quaternary structure of the large subunit of RNR, overview. Binds also mutant D57N-alpha subunit. CDP protects against inhibition
clofarabine triphosphate
-
ClFTP, an A-site rapidly binding reversible inhibitor, mechanism of inhibition via altering the quaternary structure of the large subunit of RNR, overview. Neither CDP (C site) nor dGTP (A site) had any effect on inhibition by ClFTP
Co2+
-
RNR activity chelates with copper leading to inactivation
dATP
-
inhibition of ADP reduction; inhibition of CDP reduction; inhibition of GDP reduction
deferoxamine mesylate
desferrioxamine
-
-
dGTP
-
-
dTTP
-
-
gemcitabine
-
-
Hydroxyurea
Isoquinoline-1-carboxaldehyde thiosemicarbazone
-
-
Methyl 3,4,5-trihydroxybenzoate
-
-
N-Methyl 3,4,5-trihydroxybenzamide
-
-
N6-(2-furanylmethyl)-9H-(3-C-methyl-beta-D-ribofuranosyl)adenine
-
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N6-(2-thienylmethyl)-9H-(3-C-methyl-beta-D-ribofuranosyl)adenine
-
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N6-(3-pyrazolyl)-9H-(3-C-methyl-beta-D-ribofuranosyl)adenine
-
-
N6-cyclobutyl-9H-(3-C-methyl-beta-D-ribofuranosyl)adenine
-
-
N6-cycloheptyl-9H-(3-C-methyl-beta-D-ribofuranosyl)adenine
-
-
N6-endo-norbonyl-9H-(3-C-methyl-beta-D-ribofuranosyl)adenine
-
-
N6-phenyl-9H-(3-C-methyl-beta-D-ribofuranosyl)adenine
-
-
nicotinohydroxamic acid
-
0.8 mM, 50% inhibition
phenylacetohydroxamic acid
-
1 mM, 50% inhibition
picolinohydroxamic acid
-
0.5 mM, 50% inhibition
Pyridine-2-carboxaldehyde thiosemicarbazone
-
-
triapine
[bis(2-acetylpyridine N,N-dimethylthiosemicarbazonato)-N,N,S-gallium(III)] hexafluorophosphate
-
-
[bis(2-acetylpyridine N,N-dimethylthiosemicarbazonato)-N,N,S-iron(III)] hexafluorophosphate
-
-
[bis(2-acetylpyridine N,N-dimethylthiosemicarbazonato)-N,N,S-iron(III)] tetrachloroferrate(III)
-
-
[bis(2-acetylpyridine N-pyrrolidinylthiosemicarbazonato)-N,N,S-gallium(III)] hexafluorophosphate
-
-
[bis(2-acetylpyridine N-pyrrolidinylthiosemicarbazonato)-N,N,S-iron(III)] hexafluorophosphate
-
-
[bis(2-acetylpyridine N-pyrrolidinylthiosemicarbazonato)-N,N,S-iron(III)] tetrachloroferrate(III)
-
-
[bis(acetylpyrazine N,N-dimethylthiosemicarbazonato)-N,N,S-gallium(III)] hexafluorophosphate
-
-
[bis(acetylpyrazine N,N-dimethylthiosemicarbazonato)-N,N,S-iron(III)] hexafluorophosphate
-
-
[bis(acetylpyrazine N,N-dimethylthiosemicarbazonato)-N,N,S-iron(III)] tetrachloroferrate(III)
-
-
[bis(acetylpyrazine N-piperidinylthiosemicarbazonato)-N,N,S-gallium(III)] hexafluorophosphate
-
-
[bis(acetylpyrazine N-piperidinylthiosemicarbazonato)-N,N,S-iron(III)] hexafluorophosphate
-
-
[bis(acetylpyrazine N-piperidinylthiosemicarbazonato)-N,N,S-iron(III)] tetrachloroferrate(III)
-
-
[bis(acetylpyrazine N-pyrrolidinylthiosemicarbazonato)-N,N,S-gallium(III)] hexafluorophosphate
-
-
[bis(acetylpyrazine N-pyrrolidinylthiosemicarbazonato)-N,N,S-iron(III)] hexafluorophosphate
-
-
[bis(acetylpyrazine N-pyrrolidinylthiosemicarbazonato)-N,N,S-iron(III)] tetrachloroferrate(III)
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dCTP
-
stimulation of UDP reduction
P53
-
activates, required
additional information
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00004
clofarabine
-
pH not specified in the publication, temperature not specified in the publication
additional information
additional information
-
inhibition kinetics of clofarabine di- and triphosphates, overview
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.011
(2E)-2-(anthracen-9-ylmethylidene)-N-hydroxyhydrazinecarboximidamide
Homo sapiens
-
pH not specified in the publication, temperature not specified in the publication
0.5
2-furan-3-ylbenzaldehyde N-(4-hydroxyphenyl)thiosemicarbazone
Homo sapiens
-
above, pH 7.2, 37C
0.5
2-furan-3-ylbenzaldehyde N-phenylthiosemicarbazone
Homo sapiens
-
above, pH 7.2, 37C
0.5
2-hydroxybenzaldehyde N-(4-chlorophenyl)thiosemicarbazone
Homo sapiens
-
above, pH 7.2, 37C
0.5
2-hydroxybenzaldehyde N-phenylthiosemicarbazone
Homo sapiens
-
above, pH 7.2, 37C
0.5
2-thiophen-2-ylbenzaldehyde N-(4-chlorophenyl)thiosemicarbazone
Homo sapiens
-
above, pH 7.2, 37C
0.5
2-thiophen-2-ylbenzaldehyde N-phenylthiosemicarbazone
Homo sapiens
-
above, pH 7.2, 37C
0.5
4-hydroxy-3-methoxybenzaldehyde N-(4-chlorophenyl)thiosemicarbazone
Homo sapiens
-
above, pH 7.2, 37C
0.5
4-hydroxy-3-methoxybenzaldehyde N-phenylthiosemicarbazone
Homo sapiens
-
above, pH 7.2, 37C
0.047
4-hydroxybenzaldehyde N-(2-chlorophenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.029
4-hydroxybenzaldehyde N-(2-hydroxyphenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.042
4-hydroxybenzaldehyde N-(2-methoxyphenyl)thiosemicarbazone
Homo sapiens
-
above, pH 7.2, 37C
0.037
4-hydroxybenzaldehyde N-(2-methylphenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.039
4-hydroxybenzaldehyde N-(2-nitrophenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.029
4-hydroxybenzaldehyde N-(3-chlorophenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.029
4-hydroxybenzaldehyde N-(3-hydroxyphenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.03
4-hydroxybenzaldehyde N-(3-methylphenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.287
4-hydroxybenzaldehyde N-(4-chlorophenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.043
4-hydroxybenzaldehyde N-(4-hydroxyphenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.014
4-hydroxybenzaldehyde N-(4-methylphenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.045
4-hydroxybenzaldehyde N-(4-nitrophenyl)thiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
0.242
4-hydroxybenzaldehyde N-phenylthiosemicarbazone
Homo sapiens
-
pH 7.2, 37C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
-
ligand binding assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
a colon adenocarcinoma cell line
Manually annotated by BRENDA team
-
small cell lung carcinoma
Manually annotated by BRENDA team
-
a myelogenous leukemia cell line
Manually annotated by BRENDA team
-
nasopharyngeal carcinoma cells, a gemcitabine-resistant cell line
Manually annotated by BRENDA team
-
Molt 4F cells
Manually annotated by BRENDA team
-
a breast carcinoma cell line
Manually annotated by BRENDA team
-
neuroepithelioma cell
Manually annotated by BRENDA team
additional information
-
RNR expression in small cell lung cancer cell lines, overview
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
enzyme may be specifically associated with mitochondria
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
78000
-
2 * 160000, subunit R1, + 2 * 78000, subunit R2, the catalytic active enzyme forms a dimer of homodimers
100000
-
alpha,beta, 1 * 100000 + 1 * 100000, Molt F4 lymphoblast cells
160000
-
2 * 160000, subunit R1, + 2 * 78000, subunit R2, the catalytic active enzyme forms a dimer of homodimers
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
alpha,beta, 1 * 100000 + 1 * 100000, Molt F4 lymphoblast cells
multimer
-
alphanbetan multi-subunit protein complex consisting of subunit types RR1 and RR2, the alpha or RR1 subunit contains the catalytic C site and two allosteric sites, while the beta or RR2 subunit houses a stable tyrosyl free radical that is transferred some 35 A to the catalytic site to initiate radical-based chemistry on the substrate
oligomer
-
RNRs are composed of alpha- and beta-subunits that form active (alpha)n(beta)m, with n or m being 2 or 6, complexes. Subunit alpha binds NDP substrates, i.e. CDP, UDP, ADP, and GDP, C site, as well as ATP and dNTPs, i.e. dATP, dGTP, TTP, allosteric effectors that control enzyme activity (A site) and substrate specificity, S site
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant His6-tagged hp53R2, sitting drop vapor diffusion method, at 25C, 0.002 ml of 4.5 mg/ml protein in 20 mM Tris, pH 7.5, are mixed with 150 mM NaCl and 0.002 ml of precipitant solution containing 0.1 M sodium citrate, pH 6.45, 1.3 M Li2SO4, and 0.5 M (NH4)2SO4, reservoir volume is 0.250 ml, 7-14 days, addition of ferrous ammonium sulfate of 5 mM 1 h prior to harvesting, X-ray diffraction structure determination and analysis at 2.6 A resolution
-
subunit RR1 in complex with TTP, dATP, TTP/GDP, TTP/ATP, and TTP/dATP, 1. TTP bound at the S-site, 2. dATP bound at the S-site, 3. TTP bound at the S-site and GDP at the C-site, 4. TTP bound at the S-site and ATP at the A-site, and 5. TTP bound at the S-site and dATP at the A-site, X-ray diffraction structure determination and analysis at resolutions of 2.4 A, 2.3 A, 3.2 A, 3.1 A, and 3.1 A, respectively
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged alpha and beta subunits by affinity chromatography
-
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
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recombinant His6-tagged subunits hRRM1 and hRRM2 from Escherichia coli strain BL21(DE3)
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recombinant His6-tagged subunits M2, p53R2, and M1 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression of His-tagged alpha and beta subunits and of His-tagged mutant alpha-subunit
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expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)
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expression of His6-tagged subunits hRRM1 and hRRM2 in Escherichia coli strain BL21(DE3)
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expression of His6-tagged subunits M2, p53R2, and M1 in Escherichia coli strain BL21(DE3)
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K95E
mutation in small subunit M2, results in dimer disassembly and enzyme activity inhibition. Mutant is capable of generating the diiron and tyrosyl radical cofactor, but the disassembly of the M2 dimer reduces its interaction with the large subunit M1. The transfection of the wild-type M2 but not the K95E mutant rescues theG1/S phase cell cycle arrest and cell growth inhibition caused by the siRNA knockdown of M2
K95E/E98K
charge-exchanging double mutation, recovers the dimerization and activity lost in mutant K95E
D16R
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site-directed mutagenesis, the mutant retains 55% of wild-type activity for CDP reduction, and 67% for ADP reduction, it is not inhibited and does not form hexamers at physiologically relevant dATP concentrations
H2E
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site-directed mutagenesis, the mutant retains 56% of wild-type activity for CDP reduction, and 56% for ADP reduction
additional information
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expression of subunit R2 siRNA 1284, targeting the AA(N19) sequence motif, inhibits R2 expression and active enzyme complex formation in different cell lines, it also inhibits cell growth and proloferation in vitro by blocking in the S-phase of the cell cycle, overview
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
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the enzyme is an important therapeutic target for anticancer drugs
medicine