Literature summary for 2.7.1.21 extracted from

Balzarini, J.; Van Daele, I.; Negri, A.; Solaroli, N.; Karlsson, A.; Liekens, S.; Gago, F.; Van Calenbergh, S.
Human mitochondrial thymidine kinase is selectively inhibited by 3-thiourea derivatives of beta-thymidine: identification of residues crucial for both inhibition and catalytic activity (2009), Mol. Pharmacol., 75, 1127-1136.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli Rosetta(DE3) cells as a fusion protein to glutathione transferase	Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
H13I	the mutant shows strongly increased Km compared to the wild type enzyme	Human alphaherpesvirus 1
H13I/Y127F	the mutant shows increased Km compared to the wild type enzyme	Human alphaherpesvirus 1
I59H	the mutant shows imcreased Km compared to the wild type enzyme	Homo sapiens
I59L	the mutant shows imcreased Km compared to the wild type enzyme	Homo sapiens
I59V	the mutant shows imcreased Km compared to the wild type enzyme	Homo sapiens
I59Y	the mutant shows decreased Km compared to the wild type enzyme	Homo sapiens
P12N	the mutant shows increased Km compared to the wild type enzyme	Human alphaherpesvirus 1
P12N/H13I	the mutant shows decreased Km compared to the wild type enzyme	Human alphaherpesvirus 1
P12N/H13I/Y127F	the mutant shows strongly increased Km compared to the wild type enzyme	Human alphaherpesvirus 1
P12N/Y127F	the mutant shows strongly increased Km compared to the wild type enzyme	Human alphaherpesvirus 1

Inhibitors

Inhibitors	Comment	Organism
1-(3,4-dichlorobenzyl)-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea	-	Homo sapiens
1-(3,4-dichlorobenzyl)-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea	-	Human alphaherpesvirus 1
1-(3,4-dichlorophenyl)-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea	-	Homo sapiens
1-(3,4-dichlorophenyl)-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea	-	Human alphaherpesvirus 1
1-benzyl-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea	-	Homo sapiens
1-benzyl-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea	-	Human alphaherpesvirus 1
1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(2S,5R)-2-(hydroxymethyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl]thiourea	-	Homo sapiens
1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(2S,5R)-2-(hydroxymethyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl]thiourea	-	Human alphaherpesvirus 1
1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(2S,5R)-2-(hydroxymethyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl]urea	-	Homo sapiens
1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(2S,5R)-2-(hydroxymethyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl]urea	-	Human alphaherpesvirus 1
1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea	-	Homo sapiens
1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea	-	Human alphaherpesvirus 1
1-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]-3-phenylthiourea	-	Homo sapiens
1-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]-3-phenylthiourea	-	Human alphaherpesvirus 1
3'-([[(4-chlorophenyl)carbamothioyl]amino]methyl)-3'-deoxythymidine	-	Homo sapiens
3'-([[(4-chlorophenyl)carbamothioyl]amino]methyl)-3'-deoxythymidine	-	Human alphaherpesvirus 1
3'-deoxy-3'-([[(3,4-dichlorophenyl)carbamothioyl]amino]methyl)thymidine	-	Homo sapiens
3'-deoxy-3'-([[(3,4-dichlorophenyl)carbamothioyl]amino]methyl)thymidine	-	Human alphaherpesvirus 1
3'-deoxy-3'-([[(4-methoxyphenyl)carbamothioyl]amino]methyl)thymidine	-	Homo sapiens
3'-deoxy-3'-([[(4-methoxyphenyl)carbamothioyl]amino]methyl)thymidine	-	Human alphaherpesvirus 1
3'-deoxy-3'-([[(4-methylphenyl)carbamothioyl]amino]methyl)thymidine	-	Homo sapiens
3'-deoxy-3'-([[(4-methylphenyl)carbamothioyl]amino]methyl)thymidine	-	Human alphaherpesvirus 1
3'-[([[4-(benzyloxy)phenyl]carbamothioyl]amino)methyl]-3'-deoxythymidine	-	Homo sapiens
3'-[([[4-(benzyloxy)phenyl]carbamothioyl]amino)methyl]-3'-deoxythymidine	-	Human alphaherpesvirus 1
3'-[([[4-chloro-3-(trifluoromethyl)phenyl]carbamothioyl]amino)methyl]-3'-deoxythymidine	-	Homo sapiens
3'-[([[4-chloro-3-(trifluoromethyl)phenyl]carbamothioyl]amino)methyl]-3'-deoxythymidine	-	Human alphaherpesvirus 1
5'-deoxy-5'-[(tricyclo[3.3.1.13,7]dec-1-ylcarbamothioyl)amino]thymidine	-	Homo sapiens
5'-deoxy-5'-[(tricyclo[3.3.1.13,7]dec-1-ylcarbamothioyl)amino]thymidine	-	Human alphaherpesvirus 1
5'-deoxy-5'-[[(diphenylmethyl)carbamothioyl]amino]thymidine	-	Homo sapiens
5'-deoxy-5'-[[(diphenylmethyl)carbamothioyl]amino]thymidine	-	Human alphaherpesvirus 1

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.00023	-	thymidine	mutant enzyme P12N/H13I, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Human alphaherpesvirus 1
0.00044	-	thymidine	wild type enzyme, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Human alphaherpesvirus 1
0.0039	-	thymidine	mutant enzyme P12N, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Human alphaherpesvirus 1
0.0039	-	thymidine	thymidine kinase-2 mutant enzyme I59Y, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Homo sapiens
0.0043	-	thymidine	wild type thymidine kinase-2, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Homo sapiens
0.0051	-	thymidine	thymidine kinase-2 mutant enzyme I59V, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Homo sapiens
0.0056	-	thymidine	thymidine kinase-2 mutant enzyme I59L, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Homo sapiens
0.006	-	thymidine	thymidine kinase-2 mutant enzyme I59H, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Homo sapiens
0.037	-	thymidine	mutant enzyme H13I/Y127F, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Human alphaherpesvirus 1
0.263	-	thymidine	mutant enzyme P12N/H13I/Y127F, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Human alphaherpesvirus 1
0.59	-	thymidine	mutant enzyme P12N/Y127F, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Human alphaherpesvirus 1
0.612	-	thymidine	mutant enzyme H13I, in 50 mM Tris/HCl, pH 8.0, 2.5 mM MgCl2, 10 mM dithiothreitol, 0.5 mM CHAPS, 3 mg/ml bovine serum albumin, at 37°C	Human alphaherpesvirus 1

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
mitochondrion	beta-thymidine kinase-2	Homo sapiens	5739	-

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-
Human alphaherpesvirus 1	P03176	-	-

Purification (Commentary)

Purification (Comment)	Organism
glutathione-Sepharose column chromatography	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + thymidine	-	Homo sapiens	ADP + thymidine 5'-phosphate	-	?
ATP + thymidine	-	Human alphaherpesvirus 1	ADP + thymidine 5'-phosphate	-	?

Synonyms

Synonyms	Comment	Organism
beta-thymidine kinase-1	-	Homo sapiens
beta-thymidine kinase-2	-	Homo sapiens
dThd kinase-1	-	Homo sapiens
dThd kinase-2	-	Homo sapiens
thymidine kinase-1	-	Homo sapiens
thymidine kinase-2	-	Homo sapiens
TK-1	-	Homo sapiens
TK-2	-	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Homo sapiens
ATP	-	Human alphaherpesvirus 1

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
0.00015	-	-	Homo sapiens	1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(2S,5R)-2-(hydroxymethyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl]thiourea
0.00042	-	-	Homo sapiens	1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(2S,5R)-2-(hydroxymethyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl]urea
0.00043	-	-	Homo sapiens	3'-[([[4-(benzyloxy)phenyl]carbamothioyl]amino)methyl]-3'-deoxythymidine
0.00047	-	-	Homo sapiens	3'-[([[4-chloro-3-(trifluoromethyl)phenyl]carbamothioyl]amino)methyl]-3'-deoxythymidine
0.00064	-	-	Homo sapiens	3'-deoxy-3'-([[(3,4-dichlorophenyl)carbamothioyl]amino]methyl)thymidine
0.0012	-	-	Homo sapiens	3'-([[(4-chlorophenyl)carbamothioyl]amino]methyl)-3'-deoxythymidine
0.0012	-	-	Homo sapiens	3'-deoxy-3'-([[(4-methylphenyl)carbamothioyl]amino]methyl)thymidine
0.0031	-	-	Homo sapiens	3'-deoxy-3'-([[(4-methoxyphenyl)carbamothioyl]amino]methyl)thymidine
0.031	-	-	Homo sapiens	5'-deoxy-5'-[(tricyclo[3.3.1.13,7]dec-1-ylcarbamothioyl)amino]thymidine
0.038	-	-	Homo sapiens	1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea
0.041	-	-	Homo sapiens	1-(3,4-dichlorobenzyl)-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea
0.046	-	-	Human alphaherpesvirus 1	3'-[([[4-chloro-3-(trifluoromethyl)phenyl]carbamothioyl]amino)methyl]-3'-deoxythymidine
0.057	-	-	Homo sapiens	1-(3,4-dichlorophenyl)-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea
0.087	-	-	Human alphaherpesvirus 1	3'-deoxy-3'-([[(3,4-dichlorophenyl)carbamothioyl]amino]methyl)thymidine
0.099	-	-	Homo sapiens	1-benzyl-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea
0.11	-	-	Homo sapiens	5'-deoxy-5'-[[(diphenylmethyl)carbamothioyl]amino]thymidine
0.195	-	-	Human alphaherpesvirus 1	1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(2S,5R)-2-(hydroxymethyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl]thiourea
0.232	-	-	Human alphaherpesvirus 1	3'-[([[4-(benzyloxy)phenyl]carbamothioyl]amino)methyl]-3'-deoxythymidine
0.335	-	-	Human alphaherpesvirus 1	1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[(2S,5R)-2-(hydroxymethyl)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-3-yl]urea
0.365	-	-	Human alphaherpesvirus 1	1-(3,4-dichlorobenzyl)-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea
0.392	-	-	Human alphaherpesvirus 1	5'-deoxy-5'-[(tricyclo[3.3.1.13,7]dec-1-ylcarbamothioyl)amino]thymidine
0.4	-	-	Human alphaherpesvirus 1	3'-([[(4-chlorophenyl)carbamothioyl]amino]methyl)-3'-deoxythymidine
0.405	-	-	Homo sapiens	1-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]-3-phenylthiourea
0.422	-	-	Human alphaherpesvirus 1	1-(3,4-dichlorophenyl)-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea
0.444	-	-	Human alphaherpesvirus 1	3'-deoxy-3'-([[(4-methylphenyl)carbamothioyl]amino]methyl)thymidine
0.472	-	-	Human alphaherpesvirus 1	1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea
0.5	-	IC50 above 0.5 mM	Human alphaherpesvirus 1	3'-deoxy-3'-([[(4-methoxyphenyl)carbamothioyl]amino]methyl)thymidine
0.5	-	IC50 above 0.5 mM	Human alphaherpesvirus 1	1-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]-3-phenylthiourea
0.5	-	IC50 above 0.5 mM	Human alphaherpesvirus 1	5'-deoxy-5'-[[(diphenylmethyl)carbamothioyl]amino]thymidine
0.5	-	IC50 above 0.5 mM	Human alphaherpesvirus 1	1-benzyl-3-[[(2R,3S,5S)-3-hydroxy-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl]methyl]thiourea