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Literature summary for 3.5.4.42 extracted from

  • Balotra, S.; Warden, A.C.; Newman, J.; Briggs, L.J.; Scott, C.; Peat, T.S.
    X-Ray structure and mutagenesis studies of the N-isopropylammelide isopropylaminohydrolase, AtzC (2015), PLoS ONE, 10, e0137700.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.4.42

Crystallization (Commentary)

Crystallization (Comment) Organism
structure of AtzC at 1.84 A with weak inhibitor malonate bound in the active site. Malonate adopts a planar conformation in the structure and packs against residue as well as interacting with Lys65, Gln160, His219 and Asn304. Enzyme is a dimer of dimers Pseudomonas sp. ADP

Protein Variants

Protein Variants Comment Organism
D188A about 6fold increase in kcat/Km value Pseudomonas sp. ADP
D188N activity similar to wild-type Pseudomonas sp. ADP
D303A activity similar to wild-type Pseudomonas sp. ADP
D303N activity similar to wild-type Pseudomonas sp. ADP
H219A 4fold increase in kcat value Pseudomonas sp. ADP
H249A substitution of an active site histidine residue, complete loss of activity Pseudomonas sp. ADP
K65A more than 15fold increase in kcat/Km value, 30fold increase in kcat for ammelide Pseudomonas sp. ADP
K65R more than 15fold increase in kcat/Km value, 12fold increase in kcat for ammelide Pseudomonas sp. ADP
N304A slight increase in kcat for ammelide Pseudomonas sp. ADP
N304D 7fold increase in kcat for ammelide Pseudomonas sp. ADP
Q160A activity similar to wild-type Pseudomonas sp. ADP
Q160E 60% reduction in kcat value Pseudomonas sp. ADP
W309A mutation opens up the area above the ring and increases the catalytic rate by almost a factor of two Pseudomonas sp. ADP
W309F slight reduction in catalytic rate Pseudomonas sp. ADP

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3
-
 ammelide mutant Q160R, pH 7.5, 28°C Pseudomonas sp. ADP
1.6
-
 ammelide mutant H219A, pH 7.5, 28°C Pseudomonas sp. ADP
1.7
-
 ammelide mutant D303A, pH 7.5, 28°C Pseudomonas sp. ADP
1.8
-
 ammelide mutant Q160A, pH 7.5, 28°C Pseudomonas sp. ADP
1.9
-
 ammelide mutant D188N, pH 7.5, 28°C Pseudomonas sp. ADP
2.2
-
 ammelide mutant D188A, pH 7.5, 28°C Pseudomonas sp. ADP
2.4
-
 ammelide mutant K65R, pH 7.5, 28°C Pseudomonas sp. ADP
3
-
 ammelide mutant W309F, pH 7.5, 28°C Pseudomonas sp. ADP
3.1
-
 ammelide wild-type, pH 7.5, 28°C Pseudomonas sp. ADP
4.2
-
 ammelide mutant N304A, pH 7.5, 28°C Pseudomonas sp. ADP
5.6
-
 ammelide mutant W309A, pH 7.5, 28°C Pseudomonas sp. ADP
6.1
-
 ammelide mutant D303N, pH 7.5, 28°C Pseudomonas sp. ADP
6.1
-
 ammelide mutant K65A, pH 7.5, 28°C Pseudomonas sp. ADP
25
-
 ammelide mutant N304D, pH 7.5, 28°C Pseudomonas sp. ADP

Organism

Organism UniProt Comment Textmining
Pseudomonas sp. ADP O52063
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ammelide + H2O
-
 Pseudomonas sp. ADP cyanuric acid + ammonia
-
 ?
N-isopropylammelide + H2O
-
 Pseudomonas sp. ADP cyanuric acid + isopropylamine
-
 ?

Subunits

Subunits Comment Organism
tetramer crystallization data Pseudomonas sp. ADP

Synonyms

Synonyms Comment Organism
AtzC
-
 Pseudomonas sp. ADP

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
57
-
 melting temperature, mutant H249A Pseudomonas sp. ADP
60
-
 melting temperature, wild-type Pseudomonas sp. ADP
63
-
 melting temperature, mutant H219A Pseudomonas sp. ADP

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
10.4
-
 ammelide mutant Q160R, pH 7.5, 28°C Pseudomonas sp. ADP
14.1
-
 ammelide mutant D188N, pH 7.5, 28°C Pseudomonas sp. ADP
16.8
-
 ammelide mutant H219A, pH 7.5, 28°C Pseudomonas sp. ADP
21.3
-
 ammelide mutant W309F, pH 7.5, 28°C Pseudomonas sp. ADP
23
-
 ammelide mutant Q160A, pH 7.5, 28°C Pseudomonas sp. ADP
26.2
-
 ammelide wild-type, pH 7.5, 28°C Pseudomonas sp. ADP
29.3
-
 ammelide mutant N304A, pH 7.5, 28°C Pseudomonas sp. ADP
35.8
-
 ammelide mutant D303A, pH 7.5, 28°C Pseudomonas sp. ADP
42.4
-
 ammelide mutant W309A, pH 7.5, 28°C Pseudomonas sp. ADP
90.4
-
 ammelide mutant D303N, pH 7.5, 28°C Pseudomonas sp. ADP
106.5
-
 ammelide mutant D188A, pH 7.5, 28°C Pseudomonas sp. ADP
189
-
 ammelide mutant N304D, pH 7.5, 28°C Pseudomonas sp. ADP
332.2
-
 ammelide mutant K65R, pH 7.5, 28°C Pseudomonas sp. ADP
778.5
-
 ammelide mutant K65A, pH 7.5, 28°C Pseudomonas sp. ADP

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
7
-
 ammelide mutant N304A, pH 7.5, 28°C Pseudomonas sp. ADP
7.1
-
 ammelide mutant W309F, pH 7.5, 28°C Pseudomonas sp. ADP
7.4
-
 ammelide mutant D188N, pH 7.5, 28°C Pseudomonas sp. ADP
7.6
-
 ammelide mutant N304D, pH 7.5, 28°C Pseudomonas sp. ADP
7.6
-
 ammelide mutant W309A, pH 7.5, 28°C Pseudomonas sp. ADP
8.1
-
 ammelide mutant Q160R, pH 7.5, 28°C Pseudomonas sp. ADP
8.5
-
 ammelide wild-type, pH 7.5, 28°C Pseudomonas sp. ADP
11
-
 ammelide mutant H219A, pH 7.5, 28°C Pseudomonas sp. ADP
13
-
 ammelide mutant Q160A, pH 7.5, 28°C Pseudomonas sp. ADP
15
-
 ammelide mutant D303N, pH 7.5, 28°C Pseudomonas sp. ADP
21
-
 ammelide mutant D303A, pH 7.5, 28°C Pseudomonas sp. ADP
49
-
 ammelide mutant D188A, pH 7.5, 28°C Pseudomonas sp. ADP
130
-
 ammelide mutant K65A, pH 7.5, 28°C Pseudomonas sp. ADP
140
-
 ammelide mutant K65R, pH 7.5, 28°C Pseudomonas sp. ADP