show Prodrug activation by Cryptosporidium thymidine kinase
Sun, X.E.; Sharling, L.; Muthalagi, M.; Mudeppa, D.G.; Pankiewicz, K.W.; Felczak, K.; Rathod, P.K.; Mead, J.; Striepen, B.; Hedstrom, L.; J. Biol. Chem. 285, 15916-15922 (2010)
Data extracted from this reference:
Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
2.7.1.21
dCDP
dCDP activates the enzyme 2.5old
Cryptosporidium parvum
2.7.1.21
dCTP
dCTP is not a phosphate donor, but instead activates the enzyme 7fold
Cryptosporidium parvum
2.7.1.21
additional information
the enzyme is not activated by dATP or dGTP
Cryptosporidium parvum
Cloned(Commentary)
EC Number
Commentary
Organism
2.7.1.21
expressed in Escherichia coli BL21(DE3) cells
Cryptosporidium parvum
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.21
dTTP
-
Cryptosporidium parvum
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.022
-
trifluoromethylthymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.032
-
3'-azido-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.04
-
5-fluoro-2'-deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.048
-
thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.05
-
2',3'-didehydro-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.118
-
deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.14
-
ATP
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.29
-
beta-L-thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.1.21
23300
-
3 * 23300, calculated from amino acid sequence
Cryptosporidium parvum
2.7.1.21
70000
-
gel filtration
Cryptosporidium parvum
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.21
Cryptosporidium parvum
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.1.21
3'-aminothymidine-Sepharose resin column chromatography and HisTrap column chromatography
Cryptosporidium parvum
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.21
ATP + 2',3'-didehydro-2',3'-dideoxythymidine
12% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 2',3'-didehydro-2',3'-dideoxythymidine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + 3'-azido-2',3'-dideoxythymidine
60% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 3'-azido-2',3'-dideoxythymidine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + 5-bromodeoxyuridine
84% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 5-bromodeoxyuridine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + 5-chlorodeoxyuridine
110% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 5-chlorodeoxyuridine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + 5-fluoro-2'-deoxyuridine
good substrate, 120% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 5-fluoro-2'-deoxyuridine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + 5-iododeoxyuridine
89% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 5-iododeoxyuridine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + beta-L-thymidine
less than 5% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + beta-L-thymidine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + deoxycytidine
7% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + dCMP
-
-
-
?
2.7.1.21
ATP + deoxyuridine
69% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + dUMP
-
-
-
?
2.7.1.21
ATP + guanosine
1% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + GMP
-
-
-
?
2.7.1.21
ATP + thymidine
100% activity
722671
Cryptosporidium parvum
ADP + dTMP
-
-
-
?
2.7.1.21
ATP + trifluoromethylthymidine
good substrate, 130% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + trifluoromethylthymidine 5’-phosphate
-
-
-
-
2.7.1.21
dGTP + thymidine
99% activity compared to ATP
722671
Cryptosporidium parvum
dGDP + dTMP
-
-
-
?
2.7.1.21
GTP + thymidine
42% activity compared to ATP
722671
Cryptosporidium parvum
GDP + dTMP
-
-
-
?
2.7.1.21
additional information
CTP, diphosphate, uridine, adenosine, cytosine, deoxyguanosine, deoxyguanosine, dTMP, thymine 1-beta-D-arabinofuranoside, 1-beta-D-arabinofuranosylcytosine, acyclovir, and 5-iodo-2'-deoxycytidine are no substrates
722671
Cryptosporidium parvum
?
-
-
-
-
2.7.1.21
UTP + thymidine
17% activity compared to ATP
722671
Cryptosporidium parvum
UDP + dTMP
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
2.7.1.21
trimer
3 * 23300, calculated from amino acid sequence
Cryptosporidium parvum
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.007
-
beta-L-thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.03
-
2',3'-didehydro-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.17
-
3'-azido-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.24
-
5-fluoro-2'-deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.28
-
ATP
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.28
-
thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.36
-
trifluoromethylthymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.53
-
deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
2.7.1.21
dCDP
dCDP activates the enzyme 2.5old
Cryptosporidium parvum
2.7.1.21
dCTP
dCTP is not a phosphate donor, but instead activates the enzyme 7fold
Cryptosporidium parvum
2.7.1.21
additional information
the enzyme is not activated by dATP or dGTP
Cryptosporidium parvum
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.21
expressed in Escherichia coli BL21(DE3) cells
Cryptosporidium parvum
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.21
dTTP
-
Cryptosporidium parvum
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.022
-
trifluoromethylthymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.032
-
3'-azido-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.04
-
5-fluoro-2'-deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.048
-
thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.05
-
2',3'-didehydro-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.118
-
deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.14
-
ATP
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.29
-
beta-L-thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.1.21
23300
-
3 * 23300, calculated from amino acid sequence
Cryptosporidium parvum
2.7.1.21
70000
-
gel filtration
Cryptosporidium parvum
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.21
3'-aminothymidine-Sepharose resin column chromatography and HisTrap column chromatography
Cryptosporidium parvum
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.21
ATP + 2',3'-didehydro-2',3'-dideoxythymidine
12% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 2',3'-didehydro-2',3'-dideoxythymidine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + 3'-azido-2',3'-dideoxythymidine
60% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 3'-azido-2',3'-dideoxythymidine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + 5-bromodeoxyuridine
84% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 5-bromodeoxyuridine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + 5-chlorodeoxyuridine
110% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 5-chlorodeoxyuridine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + 5-fluoro-2'-deoxyuridine
good substrate, 120% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 5-fluoro-2'-deoxyuridine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + 5-iododeoxyuridine
89% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + 5-iododeoxyuridine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + beta-L-thymidine
less than 5% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + beta-L-thymidine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + deoxycytidine
7% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + dCMP
-
-
-
?
2.7.1.21
ATP + deoxyuridine
69% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + dUMP
-
-
-
?
2.7.1.21
ATP + guanosine
1% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + GMP
-
-
-
?
2.7.1.21
ATP + thymidine
100% activity
722671
Cryptosporidium parvum
ADP + dTMP
-
-
-
?
2.7.1.21
ATP + trifluoromethylthymidine
good substrate, 130% activity compared to thymidine
722671
Cryptosporidium parvum
ADP + trifluoromethylthymidine 5’-phosphate
-
-
-
-
2.7.1.21
dGTP + thymidine
99% activity compared to ATP
722671
Cryptosporidium parvum
dGDP + dTMP
-
-
-
?
2.7.1.21
GTP + thymidine
42% activity compared to ATP
722671
Cryptosporidium parvum
GDP + dTMP
-
-
-
?
2.7.1.21
additional information
CTP, diphosphate, uridine, adenosine, cytosine, deoxyguanosine, deoxyguanosine, dTMP, thymine 1-beta-D-arabinofuranoside, 1-beta-D-arabinofuranosylcytosine, acyclovir, and 5-iodo-2'-deoxycytidine are no substrates
722671
Cryptosporidium parvum
?
-
-
-
-
2.7.1.21
UTP + thymidine
17% activity compared to ATP
722671
Cryptosporidium parvum
UDP + dTMP
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.7.1.21
trimer
3 * 23300, calculated from amino acid sequence
Cryptosporidium parvum
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.007
-
beta-L-thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.03
-
2',3'-didehydro-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.17
-
3'-azido-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.24
-
5-fluoro-2'-deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.28
-
ATP
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.28
-
thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.36
-
trifluoromethylthymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.53
-
deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.0024
-
beta-L-thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.064
-
2',3'-didehydro-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
2.3
-
ATP
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
4.5
-
deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
4.8
-
thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
5
-
3'-azido-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
5.8
-
5-fluoro-2'-deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
6.4
-
trifluoromethylthymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.0024
-
beta-L-thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
0.064
-
2',3'-didehydro-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
2.3
-
ATP
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
4.5
-
deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
4.8
-
thymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
5
-
3'-azido-2',3'-dideoxythymidine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
5.8
-
5-fluoro-2'-deoxyuridine
at pH 7.8 and 25°C
Cryptosporidium parvum
2.7.1.21
6.4
-
trifluoromethylthymidine
at pH 7.8 and 25°C
Cryptosporidium parvum