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Literature summary for 3.5.4.33 extracted from

  • Ragone, F.L.; Spears, J.L.; Wohlgamuth-Benedum, J.M.; Kreel, N.; Papavasiliou, F.N.; Alfonzo, J.D.
    The C-terminal end of the Trypanosoma brucei editing deaminase plays a critical role in tRNA binding (2011), RNA, 17, 1296-1306.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
-
Trypanosoma brucei

Protein Variants

Protein Variants Comment 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 Comment 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 UniProt Comment Textmining
Trypanosoma brucei
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
adenine34 in tRNAVal + H2O
-
Trypanosoma brucei hypoxanthine34 in tRNAVal + NH3
-
?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
27
-
assay at Trypanosoma brucei

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment 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 Comment Organism
8
-
assay at Trypanosoma brucei

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment 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