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Literature summary for 2.7.1.21 extracted from

  • Timm, J.; Bosch-Navarrete, C.; Recio, E.; Nettleship, J.E.; Rada, H.; Gonzalez-Pacanowska, D.; Wilson, K.S.
    Structural and kinetic characterization of thymidine kinase from Leishmania major (2015), PLoS Negl. Trop. Dis., 9, e0003781.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Leishmania major

Crystallization (Commentary)

Crystallization (Comment) Organism
structures of holo forms of the enzyme, in complex with substrate dThd and the non-hydrolysable ATP-analogue P1-(5'-adenosyl)P5-(5'-thymidyl)pentaphosphate Leishmania major

Inhibitors

Inhibitors Comment Organism Structure
dTTP
-
Leishmania major
dUTP
-
Leishmania major
P1-(5'-adenosyl)-P5-(5'-thymidyl)pentaphosphate
-
Leishmania major

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0011
-
2'-deoxythymidine pH 7.4, 25°C Leishmania major
0.002
-
5-bromo-2'-deoxyuridine pH 7.4, 25°C Leishmania major
0.003
-
3'-azido-3'-deoxythymidine pH 7.4, 25°C Leishmania major
0.0057
-
5-fluoro-2'-deoxyuridine pH 7.4, 25°C Leishmania major
0.011
-
2'-deoxyuridine pH 7.4, 25°C Leishmania major
0.058
-
ATP K0.5 value, Hill coefficient 1.6, pH 7.4, 25°C Leishmania major
0.0967
-
5-hydroxymethyl-2'-deoxyuridine pH 7.4, 25°C Leishmania major
1.462
-
uridine pH 7.4, 25°C Leishmania major

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
90000
-
SEDFIT analysis Leishmania major

Organism

Organism UniProt Comment Textmining
Leishmania major Q4QC75
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.5
-
substrates 2'-deoxythymidine, ATP, pH 7.4, 25°C Leishmania major

Storage Stability

Storage Stability Organism
-80°C, stable for several weeks Leishmania major

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + 2'-deoxythymidine
-
Leishmania major ADP + 2'-deoxythymidine 5'-phosphate
-
?
ATP + 2'-deoxyuridine
-
Leishmania major ADP + 2'-deoxyuridine 5'-phosphate
-
?
ATP + 3'-azido-3'-deoxythymidine
-
Leishmania major ADP + 3'-azido-3'-deoxythymidine 5'-phosphate
-
?
ATP + 5-bromo-2'-deoxyuridine
-
Leishmania major ADP + 5-bromo-2'-deoxyuridine 5'-phosphate
-
?
ATP + 5-fluoro-2'-deoxyuridine
-
Leishmania major ADP + 5-fluoro-2'-deoxyuridine 5'-phosphate
-
?
ATP + 5-hydroxymethyl-2'-deoxyuridine
-
Leishmania major ADP + 5-hydroxymethyl-2'-deoxyuridine 5'-phosphate
-
?
ATP + uridine
-
Leishmania major ADP + uridine 5'-phosphate
-
?

Subunits

Subunits Comment Organism
tetramer 4 * 33602, calculated and MALDI-TOF Leishmania major

Synonyms

Synonyms Comment Organism
LMJF_21_1210
-
Leishmania major

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.44
-
uridine pH 7.4, 25°C Leishmania major
1.97
-
3'-azido-3'-deoxythymidine pH 7.4, 25°C Leishmania major
2.82
-
5-hydroxymethyl-2'-deoxyuridine pH 7.4, 25°C Leishmania major
3.06
-
2'-deoxyuridine pH 7.4, 25°C Leishmania major
3.47
-
5-bromo-2'-deoxyuridine pH 7.4, 25°C Leishmania major
4.66
-
5-fluoro-2'-deoxyuridine pH 7.4, 25°C Leishmania major
262
-
2'-deoxythymidine pH 7.4, 25°C Leishmania major

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Leishmania major

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.0011
-
pH 7.4, 37°C Leishmania major P1-(5'-adenosyl)-P5-(5'-thymidyl)pentaphosphate
0.0038
-
Hill coefficient 1.6, pH 7.4, 37°C Leishmania major dTTP
0.187
-
pH 7.4, 37°C Leishmania major dUTP

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.03
-
uridine pH 7.4, 25°C Leishmania major
29.1
-
5-hydroxymethyl-2'-deoxyuridine pH 7.4, 25°C Leishmania major
276
-
2'-deoxyuridine pH 7.4, 25°C Leishmania major
651
-
3'-azido-3'-deoxythymidine pH 7.4, 25°C Leishmania major
814
-
5-fluoro-2'-deoxyuridine pH 7.4, 25°C Leishmania major
1690
-
5-bromo-2'-deoxyuridine pH 7.4, 25°C Leishmania major
2380
-
2'-deoxythymidine pH 7.4, 25°C Leishmania major