Cloned (Comment) | Organism |
---|---|
- |
Escherichia coli |
tRNA nucleotidyltransferase, EC 2.7.7.25, with its HD domain possessing 2',3'-cyclic phosphodiesterase activity | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D21A | tRNA nucleotidyltransferase mutant, EC 2.7.7.25, no effect on the 2',3'-cyclic phosphodiesterase activity | Escherichia coli |
D23A | tRNA nucleotidyltransferase mutant, EC 2.7.7.25, no effect on the 2',3'-cyclic phosphodiesterase activity | Escherichia coli |
D256A | HD domain mutant without detectable 2',3'-cyclic phosphodiesterase activity | Escherichia coli |
D306A | HD domain mutant with 2',3'-cyclic phosphodiesterase activity | Escherichia coli |
H225A | HD domain mutant without detectable 2',3'-cyclic phosphodiesterase activity | Escherichia coli |
H305A | HD domain mutant without detectable 2',3'-cyclic phosphodiesterase activity | Escherichia coli |
General Stability | Organism |
---|---|
stable | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5'-AMP | - |
Escherichia coli | |
Co2+ | - |
Escherichia coli | |
Cu2+ | - |
Escherichia coli | |
additional information | not inhibited by Mg2+, Mn2+, Ca2+ or Ni2+ | Escherichia coli | |
tRNA | 10 nM, strong competitive inhibition of 2',3'-cyclic phosphodiesterase activity, inhibition is abolished by addition of Mg2+, Mn2+ or Ca2+, but not of Ni2+ | Escherichia coli | |
Zn2+ | - |
Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
diphosphate | +/- 0.004 | Escherichia coli | |
0.13 | - |
CTP | +/- 0.01 | Escherichia coli | |
0.15 | - |
NADP | +/- 0.02 | Escherichia coli | |
0.18 | - |
ATP | +/- 0.01 | Escherichia coli | |
0.19 | - |
ADP | +/- 0.02 | Escherichia coli | |
0.49 | - |
cyclic 2',3'-AMP | pH 7, 37°C | Escherichia coli | |
0.49 | - |
2',3'-cAMP | +/- 0.04 | Escherichia coli | |
0.53 | - |
CDP | +/- 0.09 | Escherichia coli | |
0.76 | - |
2'-AMP | +/- 0.13 | Escherichia coli | |
1.6 | - |
cyclic 2',3'-GMP | pH 7, 37°C | Escherichia coli | |
1.6 | - |
2',3'-cGMP | +/- 0.22 | Escherichia coli | |
6.2 | - |
p-nitrophenyl phosphate | +/- 0.46 | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | activataes with 2'-AMP, KD: 0.24 +/- 0.1 mM | Escherichia coli | |
Mg2+ | activates with 2',3'-cAMP, KD: 0.49 +/- 0.1 mM | Escherichia coli | |
Mg2+ | activates with ATP, KD ~ 0.1 mM | Escherichia coli | |
additional information | metal-independent 2',3'-cyclic phosphodiesterase activity | Escherichia coli | |
Ni2+ | hydrolysis of bis-p-nitrophenyl phosphate is strongly dependent on Ni2+ | Escherichia coli | |
Ni2+ | activates with p-nitrophenyl phosphate, KD: 6.0 +/- 0.7 uM | Escherichia coli | |
Ni2+ | activates with PPi, KD: 1.06 +/- 0.1 uM | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
cyclic 2',3'-AMP + H2O | Escherichia coli | natural substrate, the 2',3'-cyclic phosphodiesterase activity of the C-terminal HD domain of tRNA nucleotidyltransferase, EC 2.7.7.25, is involved in the repair of the 3-CCA end of tRNA | 2'-AMP | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
2',3'-cyclic phosphodiesterase activity of the C-terminal HD domain of the multifunctional enzyme tRNA nucleotidyltransferase, EC 2.7.7.25 | - |
Escherichia coli | P06961 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant tRNA nucleotidyltransferase, EC 2.7.7.25, with its HD domain possessing 2',3'-cyclic phosphodiesterase activity | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1.49 | - |
+/- 0.08, ADP | Escherichia coli |
2.36 | - |
+/- 0.10, 2',3'-cGMP | Escherichia coli |
3.01 | - |
+/- 0.06, diphosphate | Escherichia coli |
3.21 | - |
+/- 0.1, 2',3'-cAMP | Escherichia coli |
3.71 | - |
+/- 0.17, 2'-CMP | Escherichia coli |
4.03 | - |
+/- 0.20, CTP | Escherichia coli |
4.53 | - |
+/- 0.14, ATP | Escherichia coli |
5.8 | - |
+/- 0.47, CDP | Escherichia coli |
12.4 | - |
+/- 0.34, p-nitrophenyl phosphate | Escherichia coli |
17.9 | - |
+/- 0.6, NADP | Escherichia coli |
Storage Stability | Organism |
---|---|
4°C, 5% glycerol, 0.5 M NaCl, pH 7.5, no loss in activity after several months | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2',3'-cAMP + H2O | - |
Escherichia coli | 2'-AMP + 3'-AMP | - |
? | |
2',3'-cGMP + H2O | - |
Escherichia coli | 2'-GMP + 3'-GMP | - |
? | |
2'-AMP + H2O | - |
Escherichia coli | adenosine + phosphate | - |
? | |
ADP + H2O | - |
Escherichia coli | AMP + phosphate | - |
? | |
ATP + H2O | - |
Escherichia coli | ADP + phosphate | - |
? | |
bis-p-nitrophenyl phosphate + H2O | - |
Escherichia coli | ? | - |
? | |
CDP + H2O | - |
Escherichia coli | CMP + phosphate | - |
? | |
CTP + H2O | - |
Escherichia coli | CDP + phosphate | - |
? | |
cyclic 2',3'-AMP + H2O | best substrate | Escherichia coli | 2'-AMP | - |
? | |
cyclic 2',3'-AMP + H2O | natural substrate, the 2',3'-cyclic phosphodiesterase activity of the C-terminal HD domain of tRNA nucleotidyltransferase, EC 2.7.7.25, is involved in the repair of the 3-CCA end of tRNA | Escherichia coli | 2'-AMP | - |
? | |
cyclic 2',3'-CMP + H2O | low activity | Escherichia coli | 2'-CMP | - |
? | |
cyclic 2',3'-GMP + H2O | high activity | Escherichia coli | 2'-GMP | - |
? | |
diphosphate + H2O | - |
Escherichia coli | 2 phosphate | - |
? | |
additional information | not: nucleoside 3',5'-cyclic phosphate | Escherichia coli | ? | - |
? | |
NADP+ + H2O | - |
Escherichia coli | NAD+ + phosphate | - |
? | |
nucleoside 2',3'-cyclic phosphate + H2O | - |
Escherichia coli | nucleoside 2'-phosphate | - |
? | |
p-nitrophenyl phosphate + H2O | - |
Escherichia coli | p-nitrophenol + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | - |
Escherichia coli |
monomer | predominantly | Escherichia coli |
oligomer | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.031 | 0.51 | cyclic 2',3'-AMP | pH 7, 37°C | Escherichia coli | |
1.24 | - |
ADP | +/- 0.07 | Escherichia coli | |
1.97 | - |
cyclic 2',3'-GMP | pH 7, 37°C | Escherichia coli | |
1.97 | - |
2',3'-cGMP | +/- 0.08 | Escherichia coli | |
2.51 | - |
diphosphate | +/- 0.05 | Escherichia coli | |
2.94 | - |
cyclic 2',3'-GMP | pH 7, 37°C | Escherichia coli | |
3.09 | - |
2'-AMP | +/- 0.14 | Escherichia coli | |
3.36 | - |
CTP | +/- 0.17 | Escherichia coli | |
3.78 | - |
ATP | +/- 0.12 | Escherichia coli | |
4.83 | - |
CDP | +/- 0.39 | Escherichia coli | |
7.63 | - |
cyclic 2',3'-AMP | pH 7, 37°C | Escherichia coli | |
7.63 | - |
2',3'-cAMP | +/- 0.48 | Escherichia coli | |
10.3 | - |
p-nitrophenyl phosphate | +/- 0.28 | Escherichia coli | |
14.9 | - |
NADP | +/- 0.5 | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00000168 | - |
tRNA | hydrolysis of cyclic 2',3'-AMP | Escherichia coli | |
0.29 | - |
tRNA | with ATP | Escherichia coli | |
0.59 | - |
5'-AMP | - |
Escherichia coli | |
1.08 | - |
tRNA | with 2'-AMP | Escherichia coli | |
1.68 | - |
tRNA | with 2',3'-cAMP | Escherichia coli |