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Fluorescence energy transfer studies of human deoxycytidine kinase: role of cysteine 185 in the conformational changes that occur upon substrate binding

Mani, R.S.; Usova, E.V.; Cass, C.E.; Eriksson, S.; Biochemistry 45, 3534-3541 (2006)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.1.74
native and mutant proteins expressed as His-tag fusion proteins in Escherichia coli BL21(DE3)pLysS
Homo sapiens
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.7.1.74
C146A
very low activity
Homo sapiens
2.7.1.74
C185A
lower efficiency with dCyd as the substrate with ATP und UTP, nearly 2-fold higher efficiency than wild-type dCK with dAdo as substrate and UTP as the phosphate donor
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.74
0.011
-
UTP
wild type enzyme with 2'-deoxycytidine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.017
-
UTP
C185A mutant enzyme with 2'-deoxycytidine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.019
-
ATP
wild type enzyme with 2'-deoxycytidine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.02
-
UTP
C185A mutant enzyme with 2'-deoxyadenosine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.025
-
ATP
C185A mutant enzyme with 2'-deoxycytidine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.04
-
UTP
wild type enzyme with 2'-deoxyadenosine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.087
-
ATP
C185A mutant enzyme with 2'-deoxyadenosine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.097
-
ATP
wild type enzyme with 2'-deoxyadenosine as phosphate acceptor, pH 7.6
Homo sapiens
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.1.74
additional information
Homo sapiens
phosphorylates both pyrimidine and purine deoxynucleosides, including numerous nucleoside analogue anticancer and antiviral prodrugs
?
-
-
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.74
Homo sapiens
P27707
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.1.74
recombinant enzymes using His-tag
Homo sapiens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.74
ATP + 2'-deoxyadenosine
-
672137
Homo sapiens
ADP + 2'-deoxyadenosine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxycytidine
-
672137
Homo sapiens
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
?
2.7.1.74
additional information
phosphorylates both pyrimidine and purine deoxynucleosides, including numerous nucleoside analogue anticancer and antiviral prodrugs
672137
Homo sapiens
?
-
-
-
-
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.74
native and mutant proteins expressed as His-tag fusion proteins in Escherichia coli BL21(DE3)pLysS
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.7.1.74
C146A
very low activity
Homo sapiens
2.7.1.74
C185A
lower efficiency with dCyd as the substrate with ATP und UTP, nearly 2-fold higher efficiency than wild-type dCK with dAdo as substrate and UTP as the phosphate donor
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.74
0.011
-
UTP
wild type enzyme with 2'-deoxycytidine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.017
-
UTP
C185A mutant enzyme with 2'-deoxycytidine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.019
-
ATP
wild type enzyme with 2'-deoxycytidine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.02
-
UTP
C185A mutant enzyme with 2'-deoxyadenosine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.025
-
ATP
C185A mutant enzyme with 2'-deoxycytidine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.04
-
UTP
wild type enzyme with 2'-deoxyadenosine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.087
-
ATP
C185A mutant enzyme with 2'-deoxyadenosine as phosphate acceptor, pH 7.6
Homo sapiens
2.7.1.74
0.097
-
ATP
wild type enzyme with 2'-deoxyadenosine as phosphate acceptor, pH 7.6
Homo sapiens
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.1.74
additional information
Homo sapiens
phosphorylates both pyrimidine and purine deoxynucleosides, including numerous nucleoside analogue anticancer and antiviral prodrugs
?
-
-
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.74
recombinant enzymes using His-tag
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.74
ATP + 2'-deoxyadenosine
-
672137
Homo sapiens
ADP + 2'-deoxyadenosine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxycytidine
-
672137
Homo sapiens
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
?
2.7.1.74
additional information
phosphorylates both pyrimidine and purine deoxynucleosides, including numerous nucleoside analogue anticancer and antiviral prodrugs
672137
Homo sapiens
?
-
-
-
-