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Literature summary for 3.4.11.7 extracted from

  • Iturrioz, X.; Vazeux, G.; Celerier, J.; Corvol, P.; Llorens-Cortes, C.
    Histidine 450 plays a critical role in catalysis and, with Ca2+, contributes to the substrate specificity of aminopeptidase A (2000), Biochemistry, 39, 3061-3068.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
-
Mus musculus

Protein Variants

Protein Variants Comment Organism
H450F mutation markedly lowers the level of enzyme activity and changes the sensitivity to Ca2+, The EC50 for Ca2+ is 0.025 mM in the wild-type enzyme and 0.279 mM in the mutant enzyme. 15.5fold increase of KI-value for glutamate-thiol in presence of 0.025 mM Ca2+, 18.9fold increase of KI-value for glutamate-thiol in presence of 4 mM Ca2+, 91.8fold increase of Ki-value for glutamate phosphonic acid in presence of 0.025 mM Ca2+, 22.1fold increase of Ki-value for glutamate phosphonic acid in presence of 4 mM Ca2+, 2.5fold increase of Ki-value for lysine-thiol in presence of 0.025 mM Ca2+, 2.76fold increase of Ki-value for lysine-thiol in presence of 4 mM Ca2+. Mutant enzyme shows a 1150fold decrease in the ratio of turnover number to Km-value for alpha-L-Glu-beta-naphthylamide in presence of 0.025 mM Ca2+. Mutant enzyme shows a 571fold decrease in the ratio of turnover number to Km-value for alpha-L-Glu-beta-naphthylamide in presence of 5 mM Ca2+ Mus musculus

Inhibitors

Inhibitors Comment Organism Structure
glutamate phosphonic acid
-
Mus musculus
glutamate-thiol
-
Mus musculus
lysine-thiol
-
Mus musculus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.171
-
alpha-L-Glu-beta-naphthylamide pH 7.4, 37°C, wild-type enzyme, in presence of 4 mM Ca2+ Mus musculus
0.432
-
alpha-L-Glu-beta-naphthylamide pH 7.4, 37°C, wild-type enzyme, in presence of 0.025 mM Ca2+ Mus musculus
0.697
-
alpha-L-Glu-beta-naphthylamide pH 7.4, 37°C, wild-type enzyme, without Ca2+ Mus musculus
4.383
-
alpha-L-Glu-beta-naphthylamide pH 7.4, 37°C, mutant enzyme H450F enzyme, in presence of 4 mM Ca2+ Mus musculus
5.94
-
alpha-L-Glu-beta-naphthylamide pH 7.4, 37°C, mutant enzyme H450F, in presence of 0.025 mM Ca2+ Mus musculus

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ 7.3fold increase of ratio of turnover number to Km-value for the wild-type enzyme. Activity of mutant enzyme H450F is not measurable in absence of Ca2+. His450 together with Ca2+ may contribute to substrate specificity Mus musculus

Organism

Organism UniProt Comment Textmining
Mus musculus P16406
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Mus musculus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alpha-L-Glu-beta-naphthylamide + H2O
-
Mus musculus L-Glu + 2-naphthylamine
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.8
-
alpha-L-Glu-beta-naphthylamide pH 7.4, 37°C, mutant enzyme H450F, in presence of 0.025 mM Ca2+ Mus musculus
7.5
-
alpha-L-Glu-beta-naphthylamide pH 7.4, 37°C, mutant enzyme H450F enzyme, in presence of 4 mM Ca2+ Mus musculus
93
-
alpha-L-Glu-beta-naphthylamide pH 7.4, 37°C, wild-type enzyme, without Ca2+ Mus musculus
149
-
alpha-L-Glu-beta-naphthylamide pH 7.4, 37°C, wild-type enzyme, in presence of 0.025 mM Ca2+ Mus musculus
166
-
alpha-L-Glu-beta-naphthylamide pH 7.4, 37°C, wild-type enzyme, in presence of 4 mM Ca2+ Mus musculus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000038
-
glutamate phosphonic acid pH 7.4, 37°C, wild-type enzyme, 4 mM Ca2+ Mus musculus
0.00012
-
glutamate-thiol pH 7.4, 37°C, wild-type enzyme, 4 mM Ca2+ Mus musculus
0.00036
-
glutamate-thiol pH 7.4, 37°C, wild-type enzyme, 0.025 mM Ca2+ Mus musculus
0.00046
-
lysine-thiol pH 7.4, 37°C, wild-type enzyme, Ca2+ Mus musculus
0.00047
-
lysine-thiol pH 7.4, 37°C, wild-type enzyme, 0.025 mM Ca2+ Mus musculus
0.00061
-
glutamate phosphonic acid pH 7.4, 37°C, wild-type enzyme, 0.025 mM Ca2+ Mus musculus
0.00084
-
glutamate phosphonic acid pH 7.4, 37°C, mutant enzyme H450F, 4 mM Ca2+ Mus musculus
0.0012
-
lysine-thiol pH 7.4, 37°C, mutant enzyme H450F, 0.025 mM Ca2+ Mus musculus
0.00167
-
glutamate-thiol pH 7.4, 37°C, wild-type enzyme, no Ca2+ Mus musculus
0.00227
-
glutamate-thiol pH 7.4, 37°C, mutant enzyme H450F, 4 mM Ca2+ Mus musculus
0.00287
-
glutamate phosphonic acid pH 7.4, 37°C, wild-type enzyme, no Ca2+ Mus musculus
0.00524
-
glutamate-thiol pH 7.4, 37°C, mutant enzyme H450F, 0.025 mM Ca2+ Mus musculus
0.0072
-
lysine-thiol pH 7.4, 37°C, wild-type enzyme, 4 mM Ca2+ Mus musculus
0.0199
-
lysine-thiol pH 7.4, 37°C, mutant enzyme H450F, 4 mM Ca2+ Mus musculus
0.056
-
glutamate phosphonic acid pH 7.4, 37°C, mutant enzyme H450F, 0.025 mM Ca2+ Mus musculus