Cloned (Comment) | Organism |
---|---|
gene Rc1, expression of soluble unmodified enzyme and His-tagged enzymes in Escherichia coli strain Bli5 (BL21 (DE3) pDIA17) | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
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 | Comment | Organism | Structure |
---|---|---|---|
6-methylthio-GMP | competitive versus dGMP | Rattus norvegicus | |
GMP | competitive versus dGMP | Rattus norvegicus | |
Zn2+ | - |
Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics and substrate specifiicty | Rattus norvegicus | |
0.048 | - |
dGMP | pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus | |
0.25 | - |
dAMP | pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus | |
0.45 | - |
dIMP | pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus | |
4 | - |
dCMP | pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus | |
5 | - |
dGMP | pH 7.5, 37°C, recombinant mutant E105A | Rattus norvegicus | |
15 | - |
dUMP | pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus | |
50 | - |
dTMP | above, pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | no effect by Ca2+, Mg2+, Na+, and K+ | Rattus norvegicus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
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 |
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 |
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 |
20440 | - |
monomeric enzyme, equilibrium sedimentation ultracentrifugation | Rattus norvegicus |
40520 | - |
monomeric enzyme, equilibrium sedimentation ultracentrifugation | Rattus norvegicus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
gene Rc1 | - |
Purification (Comment) | Organism |
---|---|
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 | Comment | Organism | Reaction ID |
---|---|---|---|
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 |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
fibroblast | - |
Rattus norvegicus | - |
Storage Stability | Organism |
---|---|
-20°C, purified recombinant enzyme, 1 mg/ml, at least 1 year without loss of activity | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dAMP + base | - |
Rattus norvegicus | dNMP + adenine | - |
r | |
dCMP + base | - |
Rattus norvegicus | dNMP + cytidine | - |
r | |
dGMP + base | dGMP is the best substrate | Rattus norvegicus | dNMP + guanine | - |
r | |
dIMP + base | - |
Rattus norvegicus | dNMP + hypoxanthine | - |
r | |
dTMP + base | - |
Rattus norvegicus | dNMP + thymine | - |
r | |
dUMP + base | - |
Rattus norvegicus | dNMP + uracil | - |
r | |
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 | Comment | Organism |
---|---|---|
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 |
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 |
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 | Comment | Organism |
---|---|---|
deoxynucleoside 5-monophosphate N-glycosidase | - |
Rattus norvegicus |
RCL | - |
Rattus norvegicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0066 | - |
dAMP | pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus | |
0.0066 | - |
dIMP | pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus | |
0.0178 | - |
dGMP | pH 7.5, 37°C, recombinant mutant E105A | Rattus norvegicus | |
0.0231 | - |
dCMP | pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus | |
0.0297 | - |
dGMP | pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus | |
0.158 | - |
dUMP | pH 7.5, 37°C, recombinant wild-type enzyme | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
- |
Rattus norvegicus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
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] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
6-methylthio-GMP | pH 7.5, 37°C, recombinant wild-type enzyme, versus dGMP | Rattus norvegicus | |
0.02 | - |
GMP | pH 7.5, 37°C, recombinant wild-type enzyme, versus dGMP | Rattus norvegicus |