Cloned (Comment) | Organism |
---|---|
mutant protein expressed in Escherichia coli | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapour diffusion method, mutant enzyme crystallized in complex with its substrates (-)-beta-2,3-dideoxy-3-thiacytidine and troxacitabine | Homo sapiens |
mutant C9S/C45S/C59S/C146S in complex with L-nucleosides (-)-beta-2',3'-dideoxy-3'-thiacytidine and troxacitabine. The nucleoside binding-site tolerates substrates with different chiral configurations by maintaining virtually all of the protein-ligand interactions responsible for productive substrate positioning. The pseudo-symmetry of nucleosides and nucleoside analogs in combination with their conformational flexibility allows the L- and D-enantiomeric forms to adopt similar shapes when bound to the enzyme | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
C9S/C45S/C59S/C146S | engineered protein has a significantly improved crystallization behaviour compared with the wild type protein | Homo sapiens |
C9S/C45S/C59S/C146S | mutation of the four surface-exposed Cys residues, constructued for crystallization. Mutant displays moderate kinetic differences in the catalytic efficiency when compared with wild-type | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
(-)-beta-2',3'-dideoxy-3'-thiacytidine | wild type enzyme with ATP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.0034 | - |
ATP | cosubstrate (-)-beta-2',3'-dideoxy-3'-thiacytidine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.008 | - |
(-)-beta-2',3'-dideoxy-3'-thiacytidine | C9S/C45S/C59S/C146S mutant enzyme with ATP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.008 | - |
(-)-beta-2',3'-dideoxy-3'-thiacytidine | wild type enzyme with UTP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.008 | - |
UTP | cosubstrate (-)-beta-2',3'-dideoxy-3'-thiacytidine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.0084 | - |
ATP | cosubstrate (-)-beta-2',3'-dideoxy-3'-thiacytidine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.0106 | - |
UTP | cosubstrate troxacitabine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.011 | - |
troxacitabine | wild type enzyme with UTP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.013 | - |
troxacitabine | wild type enzyme with ATP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.0132 | - |
ATP | cosubstrate troxacitabine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.0149 | - |
UTP | cosubstrate (-)-beta-2',3'-dideoxy-3'-thiacytidine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.015 | - |
(-)-beta-2',3'-dideoxy-3'-thiacytidine | C9S/C45S/C59S/C146S mutant enzyme with UTP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.0568 | - |
ATP | cosubstrate troxacitabine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.057 | - |
troxacitabine | C9S/C45S/C59S/C146S mutant enzyme with ATP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.0578 | - |
UTP | cosubstrate troxacitabine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.058 | - |
troxacitabine | C9S/C45S/C59S/C146Smutant enzyme with UTP as phosphate donor, 37°C, pH 7.5 | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | in complex with ATP or UTP | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Homo sapiens | dCK is able to phosphorylate both D- and L-nucleosides and nucleoside analogs | ? | - |
? | |
NTP + (-)-beta-2',3'-dideoxy-3'-thiacytidine | Homo sapiens | activation of the clinically relevant substrate | NDP + (-)-beta-2',3'-dideoxy-3'-thiacytidine 5'-phosphate | - |
? | |
NTP + troxacitabine | Homo sapiens | activation of the clinically relevant substrate | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Homo sapiens | P27707 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant protein from Escherichia coli | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + (-)-beta-2',3'-dideoxy-3'-thiacytidine | - |
Homo sapiens | ADP + (-)-beta-2',3'-dideoxy-3'-thiacytidine 5'-phosphate | - |
? | |
ATP + (-)-beta-2',3'-dideoxy-3'-thiacytidine | i.e. lamivudine | Homo sapiens | ADP + (-)-beta-2',3'-dideoxy-3'-thiacytidine 5'-phosphate | - |
? | |
ATP + D-2'-deoxycytidine | - |
Homo sapiens | ADP + D-2'-deoxycytidine 5'-phosphate | - |
? | |
ATP + L-2'-deoxycytidine | - |
Homo sapiens | ADP + L-2'-deoxycytidine 5'-phosphate | - |
? | |
ATP + troxacitabine | - |
Homo sapiens | ADP + troxacitabine 5'-phosphate | - |
? | |
additional information | dCK is able to phosphorylate both D- and L-nucleosides and nucleoside analogs | Homo sapiens | ? | - |
? | |
NTP + (-)-beta-2',3'-dideoxy-3'-thiacytidine | activation of the clinically relevant substrate | Homo sapiens | NDP + (-)-beta-2',3'-dideoxy-3'-thiacytidine 5'-phosphate | - |
? | |
NTP + troxacitabine | activation of the clinically relevant substrate | Homo sapiens | ? | - |
? | |
UTP + (-)-beta-2',3'-dideoxy-3'-thiacytidine | - |
Homo sapiens | UDP + (-)-beta-2',3'-dideoxy-3'-thiacytidine 5'-phosphate | - |
? | |
UTP + D-2'-deoxycytidine | - |
Homo sapiens | UDP + D-2'-deoxycytidine 5'-phosphate | - |
? | |
UTP + L-2'-deoxycytidine | - |
Homo sapiens | UDP + L-2'-deoxycytidine 5'-phosphate | - |
? | |
UTP + troxacitabine | - |
Homo sapiens | UDP + troxacitabine 5'-phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
dCK | - |
Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
UTP | cosubstrate L-2'-deoxycytidine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.012 | - |
ATP | cosubstrate L-2'-deoxycytidine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.028 | - |
(-)-beta-2',3'-dideoxy-3'-thiacytidine | C9S/C45S/C59S/C146S mutant enzyme with ATP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.028 | - |
ATP | cosubstrate (-)-beta-2',3'-dideoxy-3'-thiacytidine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.03 | - |
(-)-beta-2',3'-dideoxy-3'-thiacytidine | wild type enzyme with ATP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.03 | - |
ATP | cosubstrate (-)-beta-2',3'-dideoxy-3'-thiacytidine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.033 | - |
ATP | cosubstrate D-2'-deoxycytidine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.036 | - |
ATP | cosubstrate L-2'-deoxycytidine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.042 | - |
UTP | cosubstrate L-2'-deoxycytidine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.049 | - |
UTP | cosubstrate D-2'-deoxycytidine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.072 | - |
(-)-beta-2',3'-dideoxy-3'-thiacytidine | C9S/C45S/C59S/C146S mutant enzyme with UTP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.072 | - |
UTP | cosubstrate (-)-beta-2',3'-dideoxy-3'-thiacytidine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.095 | - |
troxacitabine | wild type enzyme with ATP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.095 | - |
ATP | cosubstrate troxacitabine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.102 | - |
(-)-beta-2',3'-dideoxy-3'-thiacytidine | wild type enzyme with UTP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.102 | - |
UTP | cosubstrate (-)-beta-2',3'-dideoxy-3'-thiacytidine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.173 | - |
ATP | cosubstrate D-2'-deoxycytidine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.179 | - |
troxacitabine | wild type enzyme with UTP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.179 | - |
UTP | cosubstrate troxacitabine, wild-type, 37°C, pH 7.5 | Homo sapiens | |
0.192 | - |
troxacitabine | C9S/C45S/C59S/C146S mutant enzyme with ATP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.192 | - |
ATP | cosubstrate troxacitabine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.217 | - |
UTP | cosubstrate D-2'-deoxycytidine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens | |
0.524 | - |
troxacitabine | C9S/C45S/C59S/C146S mutant enzyme with UTP as phosphate donor, 37°C, pH 7.5 | Homo sapiens | |
0.524 | - |
UTP | cosubstrate troxacitabine, mutant C9S/C45S/C59S/C146S, 37°C, pH 7.5 | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Homo sapiens | |
UTP | - |
Homo sapiens |