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Literature summary for 3.5.4.3 extracted from

  • Shek, R.; Hilaire, T.; Sim, J.; French, J.B.
    Structural determinants for substrate selectivity in guanine deaminase enzymes of the amidohydrolase superfamily (2019), Biochemistry, 58, 3280-3292 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.4.3

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Escherichia coli
expression in Escherichia coli Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment) Organism
GuaD retains the overall TIM barrel tertiary structure conserved among amidohydrolase enzymes. GuaD also possesses a single zinc cation with trigonal bipyrimidal coordination geometry within its active site Escherichia coli
Gud1 retains the overall TIM barrel tertiary structure conserved among amidohydrolase enzymes. Gud1 also possesses a single zinc cation with trigonal bipyrimidal coordination geometry within its active site Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
A386S 4.8fold decrease in kcat/Km value Escherichia coli
D327L no residual activity Escherichia coli
E240D no residual activity Escherichia coli
F210A 20fold decrease in kcat/Km value Escherichia coli
H237R no residual activity Escherichia coli
L305A no residual activity Escherichia coli
Q87N no residual activity Escherichia coli
W101H 8.9fold decrease in kcat/Km value Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.061
-
 guanine wild-type, pH 7.5, 25°C Saccharomyces cerevisiae
0.08
-
 guanine mutant W101H, pH 7.5, 25°C Escherichia coli
0.14
-
 guanine mutant A386S, pH 7.5, 25°C Escherichia coli
0.21
-
 guanine wild-type, pH 7.5, 25°C Escherichia coli
0.21
-
 guanine mutant F210A, pH 7.5, 25°C Escherichia coli
0.93
-
 1-methylguanine wild-type, pH 7.5, 25°C Escherichia coli
1
-
 8-Azaguanine wild-type, pH 7.5, 25°C Escherichia coli
1.2
-
 ammeline wild-type, pH 7.5, 25°C Escherichia coli
2.6
-
 ammeline wild-type, pH 7.5, 25°C Saccharomyces cerevisiae

Metals/Ions

Metals/Ions Comment Organism Structure
Zinc GuaD possesses a single zinc cation with trigonal bipyrimidal coordination geometry within its active site Escherichia coli
Zinc Gud1 possesses a single zinc cation with trigonal bipyrimidal coordination geometry within its active site Saccharomyces cerevisiae

Organism

Organism UniProt Comment Textmining
Escherichia coli P76641
-
-
Saccharomyces cerevisiae Q07729
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-methylguanine + H2O
-
 Escherichia coli 1-methylxanthine + NH3
-
 ?
8-azaguanine + H2O
-
 Escherichia coli 8-azaxanthine + NH3
-
 ?
ammeline + H2O
-
 Escherichia coli ? + NH3
-
 ?
ammeline + H2O
-
 Saccharomyces cerevisiae ? + NH3
-
 ?
guanine + H2O
-
 Escherichia coli xanthine + NH3
-
 ?
guanine + H2O
-
 Saccharomyces cerevisiae xanthine + NH3
-
 ?
additional information 9-methylguanine, 6-O-methylguanine, 6-thioguanine, and 2,6-diaminopurine, also exhibit measurable activity but turn over very slowly, i.e. below 0.001 per s Escherichia coli ?
-
 ?

Synonyms

Synonyms Comment Organism
guaD
-
 Escherichia coli
GUD1
-
 Saccharomyces cerevisiae
YgfP
-
 Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.08
-
 1-methylguanine wild-type, pH 7.5, 25°C Escherichia coli
0.13
-
 guanine mutant W101H, pH 7.5, 25°C Escherichia coli
0.14
-
 guanine mutant F210A, pH 7.5, 25°C Escherichia coli
0.39
-
 guanine mutant A386S, pH 7.5, 25°C Escherichia coli
1
-
 ammeline wild-type, pH 7.5, 25°C Escherichia coli
2.8
-
 guanine wild-type, pH 7.5, 25°C Escherichia coli
2.8
-
 ammeline wild-type, pH 7.5, 25°C Saccharomyces cerevisiae
7
-
 8-Azaguanine wild-type, pH 7.5, 25°C Escherichia coli
8.9
-
 guanine wild-type, pH 7.5, 25°C Saccharomyces cerevisiae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.086
-
 1-methylguanine wild-type, pH 7.5, 25°C Escherichia coli
0.7
-
 guanine mutant F210A, pH 7.5, 25°C Escherichia coli
0.83
-
 ammeline wild-type, pH 7.5, 25°C Escherichia coli
1.1
-
 ammeline wild-type, pH 7.5, 25°C Saccharomyces cerevisiae
1.5
-
 guanine mutant W101H, pH 7.5, 25°C Escherichia coli
2.8
-
 guanine mutant A386S, pH 7.5, 25°C Escherichia coli
7
-
 8-Azaguanine wild-type, pH 7.5, 25°C Escherichia coli
13
-
 guanine wild-type, pH 7.5, 25°C Escherichia coli
150
-
 guanine wild-type, pH 7.5, 25°C Saccharomyces cerevisiae