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Literature summary for 3.4.11.7 extracted from

  • Claperon, C.; Banegas-Font, I.; Iturrioz, X.; Rozenfeld, R.; Maigret, B.; Llorens-Cortes, C.
    Identification of threonine 348 as a residue involved in aminopeptidase A substrate specificity (2009), J. Biol. Chem., 284, 10618-10626.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.11.7

Cloned(Commentary)

Cloned (Comment) Organism
wild type and mutant enzymes are expressed in CHO-K1 cells Mus musculus

Protein Variants

Protein Variants Comment Organism
T348D the mutation leads to a modification of the hydrolysis velocity, with no change in the affinity of the recombinant enzymes for the substrate L-Glu-2-naphthylamide, either in the absence or presence of Ca2+ Mus musculus
T348S mutant with increased activity, the mutation leads to a modification of the hydrolysis velocity, with no change in the affinity of the recombinant enzymes for the substrate L-Glu-2-naphthylamide, either in the absence or presence of Ca2+ Mus musculus
T348Y mutant with strongly reduced activity, the mutation leads to a modification of the hydrolysis velocity, with no change in the affinity of the recombinant enzymes for the substrate L-Glu-2-naphthylamide, either in the absence or presence of Ca2+ Mus musculus

Inhibitors

Inhibitors Comment Organism Structure
glutamate phosphonate
-
 Mus musculus
glutamate thiol
-
 Mus musculus
lysine thiol
-
 Mus musculus
methionine thiol
-
 Mus musculus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.159
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.198
-
 L-Glu-2-naphthylamide recombinant wild type enzyme, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.258
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.282
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
1.136
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
1.481
-
 L-Glu-2-naphthylamide recombinant wild type enzyme, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
1.919
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
2.034
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane
-
 Mus musculus 16020
-

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ the presence of 4 mM Ca2+ enhances the enzymatic activity of wild type enzyme by a factor of 8 Mus musculus
Zn2+ dependent on Mus musculus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
140000
-
 minor immature form, SDS-PAGE Mus musculus
160000
-
 major mature form, SDS-PAGE Mus musculus

Organism

Organism UniProt Comment Textmining
Mus musculus P16406
-
-

Purification (Commentary)

Purification (Comment) Organism
Talon Co2+ column chromatography Mus musculus

Source Tissue

Source Tissue Comment Organism Textmining
brain
-
 Mus musculus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.25
-
 mutant enzyme T348Y, in the absence of Ca2+ Mus musculus
0.8
-
 mutant enzyme T348D, in the absence of Ca2+ Mus musculus
1.6
-
 mutant enzyme T348D, in the presence of 4 mM Ca2+ Mus musculus
1.98
-
 wild type enzyme, in the absence of Ca2+ Mus musculus
3.05
-
 mutant enzyme T348S, in the absence of Ca2+ Mus musculus
7.4
-
 mutant enzyme T348Y, in the presence of 4 mM Ca2+ Mus musculus
14.67
-
 wild type enzyme, in the presence of 4 mM Ca2+ Mus musculus
31.54
-
 mutant enzyme T348S, in the presence of 4 mM Ca2+ Mus musculus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
angiotensin II + H2O
-
 Mus musculus aspartic acid + angiotensin III
-
 ?
L-Glu-2-naphthylamide + H2O
-
 Mus musculus L-Glu + 2-naphthylamine
-
 ?

Synonyms

Synonyms Comment Organism
aminopeptidase A
-
 Mus musculus
APA
-
 Mus musculus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2 8 L-Glu-2-naphthylamide recombinant mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
40
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
92
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
167
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
256
-
 L-Glu-2-naphthylamide recombinant wild type enzyme, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
292
-
 L-Glu-2-naphthylamide recombinant wild type enzyme, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
575
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
640
-
 L-Glu-2-naphthylamide recombinant mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0001
-
 lysine thiol mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00019
-
 methionine thiol mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00022
-
 lysine thiol wild type enzyme, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00026
-
 methionine thiol mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00026
-
 lysine thiol mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00029
-
 methionine thiol mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00038
-
 glutamate thiol mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.00058
-
 methionine thiol wild type enzyme, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00085
-
 glutamate thiol mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.0009
-
 lysine thiol mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00103
-
 glutamate thiol mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.0011
-
 glutamate thiol wild type enzyme, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00194
-
 glutamate thiol mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00197
-
 glutamate thiol mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.002
-
 glutamate thiol wild type enzyme, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.00206
-
 glutamate thiol mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+ Mus musculus
0.00275
-
 lysine thiol mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.00298
-
 lysine thiol wild type enzyme, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.00307
-
 methionine thiol mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.0036
-
 lysine thiol mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.00394
-
 lysine thiol mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.00403
-
 methionine thiol mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.00667
-
 methionine thiol mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus
0.0084
-
 methionine thiol wild type enzyme, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+ Mus musculus