BRENDA - Enzyme Database

Biogenesis of 2-agmatinylcytidine catalyzed by the dual protein and RNA kinase TiaS

Terasaka, N.; Kimura, S.; Osawa, T.; Numata, T.; Suzuki, T.; Nat. Struct. Mol. Biol. 18, 1268-1274 (2011)

Data extracted from this reference:

Engineering
EC Number
Amino acid exchange
Commentary
Organism
6.3.4.22
C17A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
C17S
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
C210S
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
D11A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
D8A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
D9A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
E159A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
F286A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
F301A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
I143A
reduced activity
Archaeoglobus fulgidus
6.3.4.22
I49A
reduced activity
Archaeoglobus fulgidus
6.3.4.22
K52A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
N194A
reduced activity
Archaeoglobus fulgidus
6.3.4.22
N56A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
R217A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
R369A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
R44A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
R54A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
T18A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
T18D
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
T53A
complete loss of activity
Archaeoglobus fulgidus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
6.3.4.22
alpha,beta-methyleneadenosine 5'-triphosphate
-
Archaeoglobus fulgidus
6.3.4.22
beta,gamma-methyleneadenosine 5'-triphosphate
-
Archaeoglobus fulgidus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.3.4.22
0.00029
-
[tRNAIle2]-cytidine34
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
0.00087
-
agmatine
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
0.006
-
ATP
pH 8.8, 70°C
Archaeoglobus fulgidus
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
6.3.4.22
KCl
optimal concentration: 100 mM
Archaeoglobus fulgidus
6.3.4.22
Mg2+
optimal concentration: 2-5 mM
Archaeoglobus fulgidus
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
6.3.4.22
ATP + agmatine + [tRNAIle2]-cytidine34
Archaeoglobus fulgidus
the enzyme catalyzes the essential modification at the anticodon wobble position (position 34) of the AUA-codon of tRNAIle
[tRNAIle2]-2-agmatinylcytidine34 + AMP + diphosphate
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
6.3.4.22
Archaeoglobus fulgidus
O28025
-
-
Posttranslational Modification
EC Number
Posttranslational Modification
Commentary
Organism
6.3.4.22
phosphoprotein
the Thr18-Cyt34 kinase domain autophosphorylates the Thr18 of the enzyme
Archaeoglobus fulgidus
Purification (Commentary)
EC Number
Commentary
Organism
6.3.4.22
-
Archaeoglobus fulgidus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.3.4.22
ATP + agmatine + [tRNAIle2]-cytidine34
the enzyme catalyzes the essential modification at the anticodon wobble position (position 34) of the AUA-codon of tRNAIle
720531
Archaeoglobus fulgidus
[tRNAIle2]-2-agmatinylcytidine34 + AMP + diphosphate
-
-
-
?
6.3.4.22
ATP + agmatine + [tRNAIle2]-cytidine34
the precursor form of tRNAIle bearing an unmodified cytidine at the wobble position (C34) is aminoacylated with methionine (Met) and decodes the AUG codon.The Thr18-Cyt34 kinase domain first hydrolyzes ATP into AMP and diphosphate, then phosphorylates the C2 position of cytidine34 with the gamma-phosphate. Next, the amino group of agmatine attacks this position to release the phosphate and form 2-agmatinylcytidine. The Thr18-Cyt34 kinase domain also autophosphorylates the Thr18 of the enzyme, which may be involved in 2-agmatinylcytidine formation. Catalytic efficiency (kcat/Km) with GTP, UTP or CTP as substrate is less than 1% compared to ATP
720531
Archaeoglobus fulgidus
[tRNAIle2]-2-agmatinylcytidine34 + AMP + diphosphate
-
-
-
?
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
6.3.4.22
70
-
assay at
Archaeoglobus fulgidus
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.3.4.22
0.006
-
agmatine
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
0.0062
-
ATP
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
0.0065
-
[tRNAIle2]-cytidine34
pH 8.8, 70°C
Archaeoglobus fulgidus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.3.4.22
8.8
-
assay at
Archaeoglobus fulgidus
IC50 Value
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
6.3.4.22
0.00037
-
pH 8.8, 70°C
Archaeoglobus fulgidus
alpha,beta-methyleneadenosine 5'-triphosphate
6.3.4.22
0.00079
-
pH 8.8, 70°C
Archaeoglobus fulgidus
beta,gamma-methyleneadenosine 5'-triphosphate
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
6.3.4.22
C17A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
C17S
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
C210S
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
D11A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
D8A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
D9A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
E159A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
F286A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
F301A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
I143A
reduced activity
Archaeoglobus fulgidus
6.3.4.22
I49A
reduced activity
Archaeoglobus fulgidus
6.3.4.22
K52A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
N194A
reduced activity
Archaeoglobus fulgidus
6.3.4.22
N56A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
R217A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
R369A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
R44A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
R54A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
T18A
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
T18D
complete loss of activity
Archaeoglobus fulgidus
6.3.4.22
T53A
complete loss of activity
Archaeoglobus fulgidus
IC50 Value (protein specific)
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
6.3.4.22
0.00037
-
pH 8.8, 70°C
Archaeoglobus fulgidus
alpha,beta-methyleneadenosine 5'-triphosphate
6.3.4.22
0.00079
-
pH 8.8, 70°C
Archaeoglobus fulgidus
beta,gamma-methyleneadenosine 5'-triphosphate
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
6.3.4.22
alpha,beta-methyleneadenosine 5'-triphosphate
-
Archaeoglobus fulgidus
6.3.4.22
beta,gamma-methyleneadenosine 5'-triphosphate
-
Archaeoglobus fulgidus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
6.3.4.22
0.00029
-
[tRNAIle2]-cytidine34
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
0.00087
-
agmatine
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
0.006
-
ATP
pH 8.8, 70°C
Archaeoglobus fulgidus
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
6.3.4.22
KCl
optimal concentration: 100 mM
Archaeoglobus fulgidus
6.3.4.22
Mg2+
optimal concentration: 2-5 mM
Archaeoglobus fulgidus
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
6.3.4.22
ATP + agmatine + [tRNAIle2]-cytidine34
Archaeoglobus fulgidus
the enzyme catalyzes the essential modification at the anticodon wobble position (position 34) of the AUA-codon of tRNAIle
[tRNAIle2]-2-agmatinylcytidine34 + AMP + diphosphate
-
-
?
Posttranslational Modification (protein specific)
EC Number
Posttranslational Modification
Commentary
Organism
6.3.4.22
phosphoprotein
the Thr18-Cyt34 kinase domain autophosphorylates the Thr18 of the enzyme
Archaeoglobus fulgidus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
6.3.4.22
-
Archaeoglobus fulgidus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
6.3.4.22
ATP + agmatine + [tRNAIle2]-cytidine34
the enzyme catalyzes the essential modification at the anticodon wobble position (position 34) of the AUA-codon of tRNAIle
720531
Archaeoglobus fulgidus
[tRNAIle2]-2-agmatinylcytidine34 + AMP + diphosphate
-
-
-
?
6.3.4.22
ATP + agmatine + [tRNAIle2]-cytidine34
the precursor form of tRNAIle bearing an unmodified cytidine at the wobble position (C34) is aminoacylated with methionine (Met) and decodes the AUG codon.The Thr18-Cyt34 kinase domain first hydrolyzes ATP into AMP and diphosphate, then phosphorylates the C2 position of cytidine34 with the gamma-phosphate. Next, the amino group of agmatine attacks this position to release the phosphate and form 2-agmatinylcytidine. The Thr18-Cyt34 kinase domain also autophosphorylates the Thr18 of the enzyme, which may be involved in 2-agmatinylcytidine formation. Catalytic efficiency (kcat/Km) with GTP, UTP or CTP as substrate is less than 1% compared to ATP
720531
Archaeoglobus fulgidus
[tRNAIle2]-2-agmatinylcytidine34 + AMP + diphosphate
-
-
-
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
6.3.4.22
70
-
assay at
Archaeoglobus fulgidus
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
6.3.4.22
0.006
-
agmatine
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
0.0062
-
ATP
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
0.0065
-
[tRNAIle2]-cytidine34
pH 8.8, 70°C
Archaeoglobus fulgidus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.3.4.22
8.8
-
assay at
Archaeoglobus fulgidus
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
6.3.4.22
1
-
ATP
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
6.9
-
agmatine
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
22.41
-
[tRNAIle2]-cytidine34
pH 8.8, 70°C
Archaeoglobus fulgidus
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
6.3.4.22
1
-
ATP
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
6.9
-
agmatine
pH 8.8, 70°C
Archaeoglobus fulgidus
6.3.4.22
22.41
-
[tRNAIle2]-cytidine34
pH 8.8, 70°C
Archaeoglobus fulgidus