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Literature summary extracted from

  • Ghiorghi, Y.K.; Zeller, K.I.; Dang, C.V.; Kaminski, P.A.
    The c-Myc target gene Rcl (C6orf108) encodes a novel enzyme, deoxynucleoside 5-monophosphate N-glycosidase (2007), J. Biol. Chem., 282, 8150-8156.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.6 gene Rc1, expression of soluble unmodified enzyme and His-tagged enzymes in Escherichia coli strain Bli5 (BL21 (DE3) pDIA17) Rattus norvegicus

Protein Variants

EC Number Protein Variants Comment Organism
2.4.2.6 E105A site-directed mutagenesis, the mutant shows about 200fold reduced activity and a 170fold increased Km compared to the wild-type enzyme Rattus norvegicus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.2.6 6-methylthio-GMP competitive versus dGMP Rattus norvegicus
2.4.2.6 GMP competitive versus dGMP Rattus norvegicus
2.4.2.6 Zn2+
-
 Rattus norvegicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.2.6 additional information
-
 additional information kinetics and substrate specifiicty Rattus norvegicus
2.4.2.6 0.048
-
 dGMP pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus
2.4.2.6 0.25
-
 dAMP pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus
2.4.2.6 0.45
-
 dIMP pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus
2.4.2.6 4
-
 dCMP pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus
2.4.2.6 5
-
 dGMP pH 7.5, 37°C, recombinant mutant E105A Rattus norvegicus
2.4.2.6 15
-
 dUMP pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus
2.4.2.6 50
-
 dTMP above, pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.2.6 additional information no effect by Ca2+, Mg2+, Na+, and K+ Rattus norvegicus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.2.6 17649
-
 1 * 17649, unmodified recombinant enzyme, mass spectrometry, 1 * 19812, His-tagged recombinant enzyme, mass spectrometry, 1 * 20440, His-tagged recombinant enzyme, SDS-PAGE Rattus norvegicus
2.4.2.6 17649
-
 2 * 17649, unmodified recombinant enzyme, mass spectrometry, 2 * 17649, unmodified recombinant enzyme, mass spectrometry, 2 * 20440, His-tagged recombinant enzyme, SDS-PAGE Rattus norvegicus
2.4.2.6 19812
-
 1 * 17649, unmodified recombinant enzyme, mass spectrometry, 1 * 19812, His-tagged recombinant enzyme, mass spectrometry, 1 * 20440, His-tagged recombinant enzyme, SDS-PAGE Rattus norvegicus
2.4.2.6 20440
-
 monomeric enzyme, equilibrium sedimentation ultracentrifugation Rattus norvegicus
2.4.2.6 40520
-
 monomeric enzyme, equilibrium sedimentation ultracentrifugation Rattus norvegicus

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.6 Rattus norvegicus
-
 gene Rc1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.2.6 recombinant unmodified enzyme from Escherichia coli strain Bli5 (BL21 (DE3) pDIA17) by anion exchange chromatography and gel filtration, recombinant His-tagged enzyme by nickel affinity chromatography, over 95% purity Rattus norvegicus

Reaction

EC Number Reaction Comment Organism Reaction ID
2.4.2.6 2-deoxy-D-ribosyl-base1 + base2 = 2-deoxy-D-ribosyl-base2 + base1 Glu105, Asp-99 and Asn-172 are catalytic active site residues Rattus norvegicus
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.4.2.6 fibroblast
-
 Rattus norvegicus
-

Storage Stability

EC Number Storage Stability Organism
2.4.2.6 -20°C, purified recombinant enzyme, 1 mg/ml, at least 1 year without loss of activity Rattus norvegicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.6 dAMP + base
-
 Rattus norvegicus dNMP + adenine
-
 r
2.4.2.6 dCMP + base
-
 Rattus norvegicus dNMP + cytidine
-
 r
2.4.2.6 dGMP + base dGMP is the best substrate Rattus norvegicus dNMP + guanine
-
 r
2.4.2.6 dIMP + base
-
 Rattus norvegicus dNMP + hypoxanthine
-
 r
2.4.2.6 dTMP + base
-
 Rattus norvegicus dNMP + thymine
-
 r
2.4.2.6 dUMP + base
-
 Rattus norvegicus dNMP + uracil
-
 r
2.4.2.6 additional information RCL cleaves the N-glycosidic bond of deoxynucleoside 5'-monophosphates to yield two reaction products, deoxyribose 5-phosphate and purine or pyrimidine base Rattus norvegicus ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.4.2.6 dimer 2 * 17649, unmodified recombinant enzyme, mass spectrometry, 2 * 17649, unmodified recombinant enzyme, mass spectrometry, 2 * 20440, His-tagged recombinant enzyme, SDS-PAGE Rattus norvegicus
2.4.2.6 monomer 1 * 17649, unmodified recombinant enzyme, mass spectrometry, 1 * 19812, His-tagged recombinant enzyme, mass spectrometry, 1 * 20440, His-tagged recombinant enzyme, SDS-PAGE Rattus norvegicus
2.4.2.6 More at 0.003 mM the His-RCL is mostly monomeric, at 0.03 mM, His-RCL is mostly dimeric, at 0.01 mM His-RCL is at equilibrium between monomeric and dimeric states, overview Rattus norvegicus

Synonyms

EC Number Synonyms Comment Organism
2.4.2.6 deoxynucleoside 5-monophosphate N-glycosidase
-
 Rattus norvegicus
2.4.2.6 RCL
-
 Rattus norvegicus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.2.6 37
-
 assay at Rattus norvegicus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.2.6 0.0066
-
 dAMP pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus
2.4.2.6 0.0066
-
 dIMP pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus
2.4.2.6 0.0178
-
 dGMP pH 7.5, 37°C, recombinant mutant E105A Rattus norvegicus
2.4.2.6 0.0231
-
 dCMP pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus
2.4.2.6 0.0297
-
 dGMP pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus
2.4.2.6 0.158
-
 dUMP pH 7.5, 37°C, recombinant wild-type enzyme Rattus norvegicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.2.6 6
-
-
 Rattus norvegicus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.4.2.6 5 8 50% of maximal activity at pH 7.0 with substrate dCMP, 20% with dGMP, 10% of maximal activity at pH 8.0 with both substrates Rattus norvegicus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.4.2.6 0.01
-
 6-methylthio-GMP pH 7.5, 37°C, recombinant wild-type enzyme, versus dGMP Rattus norvegicus
2.4.2.6 0.02
-
 GMP pH 7.5, 37°C, recombinant wild-type enzyme, versus dGMP Rattus norvegicus