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Literature summary 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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.5.4.42 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.4.42 1.3
-
 ammelide mutant Q160R, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 1.6
-
 ammelide mutant H219A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 1.7
-
 ammelide mutant D303A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 1.8
-
 ammelide mutant Q160A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 1.9
-
 ammelide mutant D188N, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 2.2
-
 ammelide mutant D188A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 2.4
-
 ammelide mutant K65R, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 3
-
 ammelide mutant W309F, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 3.1
-
 ammelide wild-type, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 4.2
-
 ammelide mutant N304A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 5.6
-
 ammelide mutant W309A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 6.1
-
 ammelide mutant D303N, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 6.1
-
 ammelide mutant K65A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 25
-
 ammelide mutant N304D, pH 7.5, 28°C Pseudomonas sp. ADP

Organism

EC Number Organism UniProt Comment Textmining
3.5.4.42 Pseudomonas sp. ADP O52063
-
-

Substrates and Products (Substrate)

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

Subunits

EC Number Subunits Comment Organism
3.5.4.42 tetramer crystallization data Pseudomonas sp. ADP

Synonyms

EC Number Synonyms Comment Organism
3.5.4.42 AtzC
-
 Pseudomonas sp. ADP

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.4.42 57
-
 melting temperature, mutant H249A Pseudomonas sp. ADP
3.5.4.42 60
-
 melting temperature, wild-type Pseudomonas sp. ADP
3.5.4.42 63
-
 melting temperature, mutant H219A Pseudomonas sp. ADP

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.4.42 10.4
-
 ammelide mutant Q160R, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 14.1
-
 ammelide mutant D188N, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 16.8
-
 ammelide mutant H219A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 21.3
-
 ammelide mutant W309F, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 23
-
 ammelide mutant Q160A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 26.2
-
 ammelide wild-type, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 29.3
-
 ammelide mutant N304A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 35.8
-
 ammelide mutant D303A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 42.4
-
 ammelide mutant W309A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 90.4
-
 ammelide mutant D303N, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 106.5
-
 ammelide mutant D188A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 189
-
 ammelide mutant N304D, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 332.2
-
 ammelide mutant K65R, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 778.5
-
 ammelide mutant K65A, pH 7.5, 28°C Pseudomonas sp. ADP

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.5.4.42 7
-
 ammelide mutant N304A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 7.1
-
 ammelide mutant W309F, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 7.4
-
 ammelide mutant D188N, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 7.6
-
 ammelide mutant N304D, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 7.6
-
 ammelide mutant W309A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 8.1
-
 ammelide mutant Q160R, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 8.5
-
 ammelide wild-type, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 11
-
 ammelide mutant H219A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 13
-
 ammelide mutant Q160A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 15
-
 ammelide mutant D303N, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 21
-
 ammelide mutant D303A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 49
-
 ammelide mutant D188A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 130
-
 ammelide mutant K65A, pH 7.5, 28°C Pseudomonas sp. ADP
3.5.4.42 140
-
 ammelide mutant K65R, pH 7.5, 28°C Pseudomonas sp. ADP