BRENDA - Enzyme Database show
show all sequences of 3.5.4.33

The C-terminal end of the Trypanosoma brucei editing deaminase plays a critical role in tRNA binding

Ragone, F.L.; Spears, J.L.; Wohlgamuth-Benedum, J.M.; Kreel, N.; Papavasiliou, F.N.; Alfonzo, J.D.; RNA 17, 1296-1306 (2011)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
-
Trypanosoma brucei
Engineering
Amino acid exchange
Commentary
Organism
C-terDELTA10
mutation in subunit ADAT2, inactive mutant enzyme
Trypanosoma brucei
C-terDELTA5
mutation in subunit ADAT2, kcat/Km is 72% compared to the wild-type value
Trypanosoma brucei
E92A
mutation in subunit ADAT2, inactive mutant enzyme
Trypanosoma brucei
E92A/K216A/R217A/K218A/R219A
inactive mutant enzyme
Trypanosoma brucei
K216A
mutation in subunit ADAT2, kcat/Km is 12% compared to the wild-type value
Trypanosoma brucei
K218A
mutation in subunit ADAT2, kcat/Km is 24% compared to the wild-type value
Trypanosoma brucei
additional information
deletion of the last 10 amino acids at the C terminus of TbADAT2 abolishes tRNA binding. In addition, single alanine replacements of a string of positively charged amino acids (KRKRK) lead to binding defects that correlate with losses in enzyme activity
Trypanosoma brucei
R217A
mutation in subunit ADAT2, kcat/Km is 16% compared to the wild-type value
Trypanosoma brucei
R219A
mutation in subunit ADAT2, kcat/Km is 20% compared to the wild-type value
Trypanosoma brucei
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00075
-
adenine34 in tRNAVal
27°C, pH 8.0, wild-type enzyme
Trypanosoma brucei
0.00076
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme C-terDELTA5, mutation in subunit ADAT2
Trypanosoma brucei
0.002
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R218A
Trypanosoma brucei
0.0025
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R217A
Trypanosoma brucei
0.0028
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R219A
Trypanosoma brucei
0.003
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme K216A
Trypanosoma brucei
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Trypanosoma brucei
-
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
adenine34 in tRNAVal + H2O
-
721030
Trypanosoma brucei
hypoxanthine34 in tRNAVal + NH3
-
-
-
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
27
-
assay at
Trypanosoma brucei
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0015
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R217A
Trypanosoma brucei
0.0017
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme K216A
Trypanosoma brucei
0.002
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R218A
Trypanosoma brucei
0.0022
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R219A
Trypanosoma brucei
0.0023
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme C-terDELTA5, mutation in subunit ADAT2
Trypanosoma brucei
0.0032
-
adenine34 in tRNAVal
27°C, pH 8.0, wild-type enzyme
Trypanosoma brucei
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Trypanosoma brucei
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Trypanosoma brucei
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
C-terDELTA10
mutation in subunit ADAT2, inactive mutant enzyme
Trypanosoma brucei
C-terDELTA5
mutation in subunit ADAT2, kcat/Km is 72% compared to the wild-type value
Trypanosoma brucei
E92A
mutation in subunit ADAT2, inactive mutant enzyme
Trypanosoma brucei
E92A/K216A/R217A/K218A/R219A
inactive mutant enzyme
Trypanosoma brucei
K216A
mutation in subunit ADAT2, kcat/Km is 12% compared to the wild-type value
Trypanosoma brucei
K218A
mutation in subunit ADAT2, kcat/Km is 24% compared to the wild-type value
Trypanosoma brucei
additional information
deletion of the last 10 amino acids at the C terminus of TbADAT2 abolishes tRNA binding. In addition, single alanine replacements of a string of positively charged amino acids (KRKRK) lead to binding defects that correlate with losses in enzyme activity
Trypanosoma brucei
R217A
mutation in subunit ADAT2, kcat/Km is 16% compared to the wild-type value
Trypanosoma brucei
R219A
mutation in subunit ADAT2, kcat/Km is 20% compared to the wild-type value
Trypanosoma brucei
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00075
-
adenine34 in tRNAVal
27°C, pH 8.0, wild-type enzyme
Trypanosoma brucei
0.00076
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme C-terDELTA5, mutation in subunit ADAT2
Trypanosoma brucei
0.002
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R218A
Trypanosoma brucei
0.0025
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R217A
Trypanosoma brucei
0.0028
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R219A
Trypanosoma brucei
0.003
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme K216A
Trypanosoma brucei
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
adenine34 in tRNAVal + H2O
-
721030
Trypanosoma brucei
hypoxanthine34 in tRNAVal + NH3
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
27
-
assay at
Trypanosoma brucei
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0015
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R217A
Trypanosoma brucei
0.0017
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme K216A
Trypanosoma brucei
0.002
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R218A
Trypanosoma brucei
0.0022
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R219A
Trypanosoma brucei
0.0023
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme C-terDELTA5, mutation in subunit ADAT2
Trypanosoma brucei
0.0032
-
adenine34 in tRNAVal
27°C, pH 8.0, wild-type enzyme
Trypanosoma brucei
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Trypanosoma brucei
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.57
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme K216A
Trypanosoma brucei
0.59
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R217A
Trypanosoma brucei
0.79
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R219A
Trypanosoma brucei
1
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R218A
Trypanosoma brucei
3.03
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme C-terDELTA5, mutation in subunit ADAT2
Trypanosoma brucei
4.27
-
adenine34 in tRNAVal
27°C, pH 8.0, wild-type enzyme
Trypanosoma brucei
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.57
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme K216A
Trypanosoma brucei
0.59
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R217A
Trypanosoma brucei
0.79
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R219A
Trypanosoma brucei
1
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme R218A
Trypanosoma brucei
3.03
-
adenine34 in tRNAVal
27°C, pH 8.0, mutant enzyme C-terDELTA5, mutation in subunit ADAT2
Trypanosoma brucei
4.27
-
adenine34 in tRNAVal
27°C, pH 8.0, wild-type enzyme
Trypanosoma brucei
Other publictions for EC 3.5.4.33
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
718952
Su
A-to-I and C-to-U editing with ...
Agrobacterium tumefaciens, Aquifex aeolicus, Escherichia coli, Streptococcus pyogenes
Biochemistry
76
932-937
2011
-
-
-
-
-
-
-
-
-
-
-
4
-
9
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
719934
Spears
A single zinc ion is sufficien ...
Trypanosoma brucei
J. Biol. Chem.
286
20366-20374
2011
-
-
-
-
11
-
-
4
-
1
-
1
-
4
-
-
-
-
-
-
-
-
1
-
1
-
-
3
1
-
-
-
-
-
-
-
-
-
-
-
11
-
-
-
-
4
-
1
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
3
1
-
-
-
-
-
-
-
4
4
721030
Ragone
The C-terminal end of the Tryp ...
Trypanosoma brucei
RNA
17
1296-1306
2011
-
-
1
-
9
-
-
6
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
6
1
-
-
-
-
-
-
-
-
1
-
-
9
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
6
1
-
-
-
-
-
-
-
6
6
713269
Delannoy
Arabidopsis tRNA adenosine dea ...
Arabidopsis thaliana
Plant Cell
21
2058-2071
2009
-
-
1
-
-
-
-
-
2
-
-
1
-
7
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
721023
Karcher
Identification of the chloropl ...
Arabidopsis thaliana
RNA
15
1251-1257
2009
-
-
1
-
-
-
-
-
1
-
-
1
-
6
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
698470
Luo
Transition state structure of ...
Escherichia coli
J. Am. Chem. Soc.
130
2649-2655
2008
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
719843
Tsutsumi
Wobble inosine tRNA modificati ...
Schizosaccharomyces pombe
J. Biol. Chem.
282
33459-33465
2007
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
720892
Rubio
An adenosine-to-inosine tRNA-e ...
Trypanosoma brucei
Proc. Natl. Acad. Sci. USA
104
7821-7826
2007
-
-
-
-
-
-
-
-
-
-
-
1
-
4
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
721010
Lee
Crystal structure of the tRNA- ...
Streptococcus pyogenes
Proteins
68
1016-1019
2007
-
-
-
1
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
689140
Losey
Crystal structure of Staphyloc ...
Staphylococcus aureus
Nat. Struct. Mol. Biol.
13
153-159
2006
-
-
1
1
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
669323
Kuratani
Crystal structure of tRNA aden ...
Aquifex aeolicus
J. Biol. Chem.
280
16002-16008
2005
-
-
1
1
-
-
-
-
-
1
-
1
-
1
-
-
1
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
1
-
1
-
-
-
1
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
718485
Ku
Crystallization and preliminar ...
Streptococcus pyogenes, Streptococcus pyogenes ATCC BAA-946
Acta Crystallogr. Sect. F
61
375-357
2005
-
-
1
1
-
-
-
-
-
-
-
-
-
5
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
718839
Elias
Biochemical and structural stu ...
Agrobacterium tumefaciens, Aquifex aeolicus, Escherichia coli, Saccharomyces cerevisiae, Saccharomyces cerevisiae ATCC 204508
Biochemistry
44
12057-12065
2005
-
-
4
1
4
-
-
-
-
1
-
5
-
14
-
-
4
-
-
-
-
-
10
4
4
-
-
-
4
-
-
-
-
-
-
-
-
4
-
1
4
-
-
-
-
-
-
1
-
5
-
-
-
4
-
-
-
-
10
4
4
-
-
-
4
-
-
-
-
-
-
-
-
-
719356
Wolf
tadA, an essential tRNA-specif ...
Escherichia coli
EMBO J.
21
3841-3851
2002
-
-
1
-
1
-
-
-
-
-
1
1
-
4
-
-
1
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
1
1
-
-
-
1
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
1
1
-
-
-
721050
Gerber
An adenosine deaminase that ge ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae ATCC 204508
Science
286
1146-1149
1999
-
-
-
-
1
-
-
-
-
-
-
2
-
4
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
720220
Auxilien
Mechanism, specificity and gen ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae ATCC 204508
J. Mol. Biol.
262
437-458
1996
-
-
-
-
-
-
3
1
-
1
1
2
-
7
-
-
1
-
-
-
-
-
8
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
1
-
1
1
2
-
-
-
1
-
-
-
-
8
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-