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

BRENDA support

Literature summary for 1.17.4.1 extracted from

  • Xu, H.; Fairman, J.W.; Wijerathna, S.R.; Kreischer, N.R.; LaMacchia, J.; Helmbrecht, E.; Cooperman, B.S.; Dealwis, C.
    The structural basis for peptidomimetic inhibition of eukaryotic ribonucleotide reductase: a conformationally flexible pharmacophore (2008), J. Med. Chem., 51, 4653-4659.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
medicine the enzyme is a target for cancer therapy Saccharomyces cerevisiae

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-tagged subunits R2 and R4 in Escherichia coli strain BL21(DE3) Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment) Organism
purified enzyme, formed by recombinant subunits R2 and R4, complexed with inhibitor peptide P7 or peptide Fmoc-P6, 20 mg/ml protein in 0.1 M HEPES, pH 7.5, with 20 mM TTP, 5% glycerol, 5mM DTT, 0.1 M KCL, and 25 mm MgCl2, is mixed with a reservoir solution containing 20-25% PEG 3350, 0.2 M NaCl, and 100 mM HEPES, pH 7.5, soaking of crystals for 4 h in reservoir solution with added inhibitor peptide P6 or P7, followed by soaking of crystals in 25% PEG 3350, 0.2 M NaCl, and 100 mM HEPES, pH 7.5, supplemented with 15% glycerol, X-ray diffraction structure determination and analysis at 2.6 and 2.5 A resolution, respectively Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
additional information construction of heterochimeric enzymes as heterocomplexes containing mammalian R2 C-terminal heptapeptide P7, Ac-1FTLDADF7, and its peptidomimetic P6, 1Fmoc(Me)PhgLDChaDF7, bound to Saccharomyces cerevisiae R1, ScR1, overview Saccharomyces cerevisiae

Inhibitors

Inhibitors Comment Organism Structure
mammalian R2 C-terminal heptapeptide P7 Ac-1FTLDADF7, the inhibitor binds at bind at a contiguous site containing residues that are highly conserved among eukaryotes, binding structure, overview Saccharomyces cerevisiae
peptide P6 1Fmoc(Me)PhgLDChaDF7, the inhibitor binds at a contiguous site containing residues that are highly conserved among eukaryotes. The Fmoc group in P6 peptide forms several hydrophobic interactions that contribute to its enhanced potency in binding to ScR1, binding structure, overview Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
GDP + thioredoxin Saccharomyces cerevisiae
-
2'-deoxyGDP + thioredoxin disulfide + H2O
-
?
ribonucleoside diphosphate + thioredoxin Saccharomyces cerevisiae
-
2'-deoxyribonucleoside diphosphate + thioredoxin disulfide + H2O
-
?

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae P21524
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged subunits R2 and R4 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
GDP + thioredoxin
-
Saccharomyces cerevisiae 2'-deoxyGDP + thioredoxin disulfide + H2O
-
?
ribonucleoside diphosphate + thioredoxin
-
Saccharomyces cerevisiae 2'-deoxyribonucleoside diphosphate + thioredoxin disulfide + H2O
-
?

Subunits

Subunits Comment Organism
tetramer the enzyme activity requires formation of a complex between subunits R1 and R2 in which the R2 C-terminal peptide binds to R1 Saccharomyces cerevisiae

Synonyms

Synonyms Comment Organism
ribonucleotide reductase
-
Saccharomyces cerevisiae

Cofactor

Cofactor Comment Organism Structure
thioredoxin
-
Saccharomyces cerevisiae

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.0026
-
yeast enzyme Saccharomyces cerevisiae peptide Fmoc-P6
0.031
-
yeast enzyme Saccharomyces cerevisiae peptide P7