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Literature summary for 3.1.4.37 extracted from

  • Yakunin, A.F.; Proudfoot, M.; Kuznetsova, E.; Savchenko, A.; Brown, G.; Arrowsmith, C.H.; Edwards, A.M.
    The HD domain of the E. coli tRNA nucleotidyltransferase has 2',3'-cyclic phosphodiesterase, 2'-nucleotidase, and phosphatase activities (2004), J. Biol. Chem., 279, 36819-36827.
    View publication on PubMed

Cloned(Commentary)

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

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

General Stability Organism
stable Escherichia coli

Inhibitors

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 [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

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/ Products (Substrates)

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

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 (Commentary)

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 [micromol/min/mg]

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

Storage Stability Organism
4°C, 5% glycerol, 0.5 M NaCl, pH 7.5, no loss in activity after several months Escherichia coli

Substrates and Products (Substrate)

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

Subunits Comment Organism
dimer
-
Escherichia coli
monomer predominantly Escherichia coli
oligomer
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Escherichia coli

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Escherichia coli

Ki Value [mM]

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