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Literature summary for 3.5.4.4 extracted from

  • Ho, M.C.; Cassera, M.B.; Madrid, D.C.; Ting, L.M.; Tyler, P.C.; Kim, K.; Almo, S.C.; Schramm, V.L.
    Structural and metabolic specificity of methylthiocoformycin for malarial adenosine deaminases (2009), Biochemistry, 48, 9618-9626.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
wild type enzyme is expressed in Escherichia coli strain TOP10, mutant enzymes are expressed in Escherichia coli strain BL21-codon plus (DE3)-RIPL Plasmodium vivax

Crystallization (Commentary)

Crystallization (Comment) Organism
Plasmodium vivax ADA in a complex with 5'-methylthiocoformycin is crystallized by the sitting drop vapor diffusion method, using 25% (w/v) PEG3350, 100 mM HEPES (pH 7.5), and 0.2 M MgCl2 Plasmodium vivax

Protein Variants

Protein Variants Comment Organism
D172A the mutant shows no activity with 5-methylthioadenosine and is not inhibited by 5'-methylthiocoformycin Plasmodium vivax
D172E the mutant shows no activity with 5-methylthioadenosine and is not inhibited by 5'-methylthiocoformycin Plasmodium vivax

Inhibitors

Inhibitors Comment Organism Structure
5'-methylthiocoformycin specific transition state analogue inhibitor of Plasmodium ADA Plasmodium berghei
5'-methylthiocoformycin specific transition state analogue inhibitor of Plasmodium ADA Plasmodium cynomolgi
5'-methylthiocoformycin specific transition state analogue inhibitor of Plasmodium ADA, 5'-methylthiocoformycin is a subnanomolar inhibitor of Plasmodium falciparum ADA and demonstrates more than 20000fold selectivity relative to human ADA Plasmodium falciparum
5'-methylthiocoformycin poor inhibitor of Plasmodium gallinaceum ADA Plasmodium gallinaceum
5'-methylthiocoformycin specific transition state analogue inhibitor of Plasmodium ADA Plasmodium knowlesi
5'-methylthiocoformycin specific transition state analogue inhibitor of Plasmodium ADA Plasmodium vivax
coformycin powerful picomolar inhibitor of ADA Plasmodium berghei
coformycin powerful picomolar inhibitor of ADA Plasmodium cynomolgi
coformycin powerful picomolar inhibitor of ADA Plasmodium falciparum
coformycin powerful picomolar inhibitor of ADA Plasmodium gallinaceum
coformycin powerful picomolar inhibitor of ADA Plasmodium knowlesi
coformycin powerful picomolar inhibitor of ADA Plasmodium vivax
D-coformycin powerful picomolar inhibitor of ADA Plasmodium berghei
D-coformycin powerful picomolar inhibitor of ADA Plasmodium cynomolgi
D-coformycin powerful picomolar inhibitor of ADA Plasmodium falciparum
D-coformycin powerful picomolar inhibitor of ADA Plasmodium gallinaceum
D-coformycin powerful picomolar inhibitor of ADA Plasmodium knowlesi
D-coformycin powerful picomolar inhibitor of ADA Plasmodium vivax

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0044
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium berghei
0.0087
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium cynomolgi
0.0095
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
0.022
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium knowlesi
0.032
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium gallinaceum
0.057
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium berghei
0.06
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
0.083
-
adenosine mutant enzyme D172E, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
0.087
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium cynomolgi
0.088
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium falciparum
0.104
-
adenosine mutant enzyme D172A, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
0.115
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium falciparum
0.12
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium knowlesi

Organism

Organism UniProt Comment Textmining
Plasmodium berghei
-
-
-
Plasmodium cynomolgi
-
-
-
Plasmodium falciparum
-
-
-
Plasmodium gallinaceum
-
-
-
Plasmodium knowlesi
-
-
-
Plasmodium vivax
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Plasmodium vivax

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5'-methylthioadenosine + H2O
-
Plasmodium knowlesi 5'-methylthioinosine + NH3
-
?
5'-methylthioadenosine + H2O
-
Plasmodium falciparum 5'-methylthioinosine + NH3
-
?
5'-methylthioadenosine + H2O
-
Plasmodium berghei 5'-methylthioinosine + NH3
-
?
5'-methylthioadenosine + H2O
-
Plasmodium vivax 5'-methylthioinosine + NH3
-
?
5'-methylthioadenosine + H2O
-
Plasmodium cynomolgi 5'-methylthioinosine + NH3
-
?
adenosine + H2O
-
Plasmodium knowlesi inosine + NH3
-
?
adenosine + H2O
-
Plasmodium falciparum inosine + NH3
-
?
adenosine + H2O
-
Plasmodium berghei inosine + NH3
-
?
adenosine + H2O
-
Plasmodium vivax inosine + NH3
-
?
adenosine + H2O
-
Plasmodium gallinaceum inosine + NH3
-
?
adenosine + H2O
-
Plasmodium cynomolgi inosine + NH3
-
?
additional information Plasmodium gallinaceum ADA does not have significant activity for 5'-methylthioadenosine Plasmodium gallinaceum ?
-
?

Synonyms

Synonyms Comment Organism
ADA
-
Plasmodium knowlesi
ADA
-
Plasmodium falciparum
ADA
-
Plasmodium berghei
ADA
-
Plasmodium vivax
ADA
-
Plasmodium gallinaceum
ADA
-
Plasmodium cynomolgi

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.13
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
0.31
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium cynomolgi
0.35
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium berghei
0.51
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium knowlesi
1.8
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
1.9
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium gallinaceum
4.7
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium berghei
5.3
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium cynomolgi
5.6
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium falciparum
5.8
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium falciparum
6.8
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium knowlesi
12.9
-
adenosine mutant enzyme D172E, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
14
-
adenosine mutant enzyme D172A, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0000015
-
D-coformycin mutant enzyme D172E, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
0.0000018
-
D-coformycin mutant enzyme D172A, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
0.0000023
-
D-coformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium berghei
0.0000032
-
5'-methylthiocoformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium falciparum
0.0000034
-
D-coformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium knowlesi
0.0000047
-
D-coformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium gallinaceum
0.000007
-
D-coformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium cynomolgi
0.0000074
-
D-coformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
0.000014
-
5'-methylthiocoformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium berghei
0.000014
-
D-coformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium falciparum
0.00002
-
5'-methylthiocoformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
0.00003
-
5'-methylthiocoformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium cynomolgi
0.000048
-
5'-methylthiocoformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium knowlesi
0.029
-
5'-methylthiocoformycin wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium gallinaceum

General Information

General Information Comment Organism
metabolism ADA is a key enzyme in the pathway of hypoxanthine formation Plasmodium knowlesi
metabolism ADA is a key enzyme in the pathway of hypoxanthine formation Plasmodium falciparum
metabolism ADA is a key enzyme in the pathway of hypoxanthine formation Plasmodium berghei
metabolism ADA is a key enzyme in the pathway of hypoxanthine formation Plasmodium vivax
metabolism ADA is a key enzyme in the pathway of hypoxanthine formation Plasmodium gallinaceum
metabolism ADA is a key enzyme in the pathway of hypoxanthine formation Plasmodium cynomolgi

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
14
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
23
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium knowlesi
30
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
36
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium cynomolgi
50
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium falciparum
57
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium knowlesi
59
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium gallinaceum
61
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium cynomolgi
64
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium falciparum
79
-
5'-methylthioadenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium berghei
82
-
adenosine wild type enzyme, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium berghei
130
-
adenosine mutant enzyme D172A, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax
150
-
adenosine mutant enzyme D172E, in 100 mM Tris-HCl buffer (pH 8.0), temperature not specified in the publication Plasmodium vivax