Literature summary for 2.7.1.145 extracted from

Welin, M.; Skovgaard, T.; Knecht, W.; Zhu, C.; Berenstein, D.; Munch-Petersen, B.; Piskur, J.; Eklund, H.
Structural basis for the changed substrate specificity of Drosophila melanogaster deoxyribonucleoside kinase mutant N64D (2005), FEBS J., 272, 3733-3742.

Search for more literature for 2.7.1.145

Cloned(Commentary)

Cloned (Comment)	Organism
expression of the wild-type and mutant enzymes in enzyme-deficient Escherichia coli strain KY895	Drosophila melanogaster

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant mutant N64D in complex with dT or dTTP, counter diffusion and vapour diffusion methods, 20 mg/ml enzyme with 10 mM dT, is mixed with cyrstallization solution containing 0.15 M MES, pH 6.5, 0.3 M lithium sulfate, and 27,5% w/v PEG 2000 monomethylether, or 10 mg/ml enzyme with 5 mM dTTP is mixed with 0.1 M MES, pH 6.5, 0.16 M lithium sulfate, and 25% v/v PEG 2000 monomethylether, 2 weeks, 15°C, X-ray diffraction structure determination and analysis at 3.1 A and 2.2 A resolution, respectively	Drosophila melanogaster

Crystallization (Comment)

Organism

purified recombinant mutant N64D in complex with dT or dTTP, counter diffusion and vapour diffusion methods, 20 mg/ml enzyme with 10 mM dT, is mixed with cyrstallization solution containing 0.15 M MES, pH 6.5, 0.3 M lithium sulfate, and 27,5% w/v PEG 2000 monomethylether, or 10 mg/ml enzyme with 5 mM dTTP is mixed with 0.1 M MES, pH 6.5, 0.16 M lithium sulfate, and 25% v/v PEG 2000 monomethylether, 2 weeks, 15°C, X-ray diffraction structure determination and analysis at 3.1 A and 2.2 A resolution, respectively

Drosophila melanogaster

Protein Variants

Protein Variants	Comment	Organism
additional information	expression of the enzyme in enzyme-deficient Escherichia coli strain KY895 confers resistnce against 3'-azido-2',3'-didehydro-3'-deoxythymidine at more than 0.1 mM	Drosophila melanogaster
N45D	site-directed mutagenesis, decreased activity with natural substrates	Drosophila melanogaster
N45D/N64D	decreased activity with natural substrates, decreased feedback inhibition by dTTP compared to the wild-type enzyme	Drosophila melanogaster
N64D	site-directed mutagenesis, highly decreased activity with natural substrates, decreased feedback inhibition by dTTP compared to the wild-type enzyme	Drosophila melanogaster

Inhibitors

Inhibitors	Comment	Organism	Structure
dTTP	competitive versus ATP, feedback inhibition, binding at the active site, binding structure of mutant N64D, modelling	Drosophila melanogaster

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.0009	-	2'-deoxycytidine	recombinant mutant N45D	Drosophila melanogaster
0.0009	-	2'-deoxythymidine	recombinant mutants N45D	Drosophila melanogaster
0.0111	-	3'-azido-2',3'-didehydro-3'-deoxythymidine	recombinant mutant N64D	Drosophila melanogaster
0.0117	-	3'-azido-2',3'-didehydro-3'-deoxythymidine	recombinant mutant N45D	Drosophila melanogaster
0.0232	-	2'-deoxythymidine	recombinant mutant N64D	Drosophila melanogaster
0.118	-	2'-deoxycytidine	recombinant mutant N64D	Drosophila melanogaster
0.119	-	2'-deoxyadenosine	recombinant mutant N45D	Drosophila melanogaster
0.412	-	2'-deoxyguanosine	recombinant mutant N45D	Drosophila melanogaster
3.82	-	2'-deoxyadenosine	recombinant mutant N64D	Drosophila melanogaster
20.35	-	2'-deoxyguanosine	recombinant mutant N64D	Drosophila melanogaster

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Drosophila melanogaster

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
ATP + 2'-deoxyadenosine	Drosophila melanogaster	-	ADP + 2'-deoxyadenosine 5'-phosphate	-	?
ATP + 2'-deoxycytidine	Drosophila melanogaster	-	ADP + 2'-deoxycytidine 5'-phosphate	-	?
ATP + 2'-deoxyguanosine	Drosophila melanogaster	-	ADP + 2'-deoxyguanosine 5'-phosphate	-	?
ATP + 2'-deoxythymidine	Drosophila melanogaster	-	ADP + 2'-deoxythymidine 5'-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Drosophila melanogaster	-	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	-	Drosophila melanogaster

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + 2'-deoxyadenosine	-	Drosophila melanogaster	ADP + 2'-deoxyadenosine 5'-phosphate	-	?
ATP + 2'-deoxycytidine	-	Drosophila melanogaster	ADP + 2'-deoxycytidine 5'-phosphate	-	?
ATP + 2'-deoxyguanosine	-	Drosophila melanogaster	ADP + 2'-deoxyguanosine 5'-phosphate	-	?
ATP + 2'-deoxythymidine	-	Drosophila melanogaster	ADP + 2'-deoxythymidine 5'-phosphate	-	?
ATP + 2'-deoxythymidine	substrate binding structure of mutant N64D	Drosophila melanogaster	ADP + 2'-deoxythymidine 5'-phosphate	-	?
ATP + 3'-azido-2',3'-didehydro-3'-deoxythymidine	-	Drosophila melanogaster	ADP + 3'-azido-2',3'-didehydro-3'-deoxythymidine 5'-phosphate	-	?
ATP + aciclovir	-	Drosophila melanogaster	ADP + aciclovir phosphate	-	?

Synonyms

Synonyms	Comment	Organism
dNK	-	Drosophila melanogaster

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.03	-	3'-azido-2',3'-didehydro-3'-deoxythymidine	recombinant mutant N45D	Drosophila melanogaster
0.037	-	3'-azido-2',3'-didehydro-3'-deoxythymidine	recombinant mutant N64D	Drosophila melanogaster
0.12	-	2'-deoxyguanosine	recombinant mutant N64D	Drosophila melanogaster
0.4	-	2'-deoxyadenosine	recombinant mutant N64D	Drosophila melanogaster
0.82	-	2'-deoxythymidine	recombinant mutant N64D	Drosophila melanogaster
1.3	-	2'-deoxyguanosine	recombinant mutant N45D	Drosophila melanogaster
1.6	-	2'-deoxycytidine	recombinant mutants N45D and N64D	Drosophila melanogaster
1.83	-	2'-deoxyadenosine	recombinant mutant N45D	Drosophila melanogaster
2.6	-	2'-deoxythymidine	recombinant mutant N45D	Drosophila melanogaster

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Drosophila melanogaster

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
0.0047	-	dTTP	uncompetitive versus 2'-deoxythymidine, recombinant wild-type enzyme	Drosophila melanogaster
0.0163	-	dTTP	competitive versus ATP, recombinant wild-type enzyme	Drosophila melanogaster
0.52	-	dTTP	uncompetitive versus 2'-deoxythymidine, recombinant mutant N64D	Drosophila melanogaster
0.829	-	dTTP	competitive versus ATP, recombinant mutant N64D	Drosophila melanogaster