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Literature summary for 3.4.11.9 extracted from

  • Cottrell, G.S.; Hyde, R.J.; Lim, J.; Parsons, M.R.; Hooper, N.M.; Turner, A.J.
    Identification of critical residues in the active site of porcine membrane-bound aminopeptidase P (2000), Biochemistry, 39, 15129-15135.
    View publication on PubMed
Search for more literature for 3.4.11.9

Cloned(Commentary)

Cloned (Comment) Organism
transient expression of wild-type and mutants in COS-1 cells Sus scrofa

Protein Variants

Protein Variants Comment Organism
D449A site-directed mutagenesis, inactive mutant Sus scrofa
D449N site-directed mutagenesis, inactive mutant Sus scrofa
D460A site-directed mutagenesis, inactive mutant Sus scrofa
D460N site-directed mutagenesis, inactive mutant Sus scrofa
E554A site-directed mutagenesis, inactive mutant Sus scrofa
E554Q site-directed mutagenesis, inactive mutant Sus scrofa
E568A site-directed mutagenesis, inactive mutant Sus scrofa
E568Q site-directed mutagenesis, low expression level in COS-1 cells, inactive mutant Sus scrofa
E588A site-directed mutagenesis, 66% remaining activity compared to the wild-type enzyme with substrate bradykinin Sus scrofa
E588Q site-directed mutagenesis, 83% remaining activity compared to the wild-type enzyme with substrate bradykinin Sus scrofa
H429K site-directed mutagenesis, inactive mutant Sus scrofa
H429L site-directed mutagenesis, inactive mutant Sus scrofa
H519K site-directed mutagenesis, 2.3% remaining activity compared to the wild-type enzyme with substrate bradykinin, increased Km, reduced kcat Sus scrofa
H519L site-directed mutagenesis, 73.8% remaining activity compared to the wild-type enzyme with substrate bradykinin, increased Km Sus scrofa
H523K site-directed mutagenesis, inactive mutant Sus scrofa
H523L site-directed mutagenesis, inactive mutant Sus scrofa
H532K site-directed mutagenesis, inactive mutant Sus scrofa
H532L site-directed mutagenesis, very low expression level in COS-1 cells, protein degradation, inactive mutant Sus scrofa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.058
-
 bradykinin 37°C, wild-type enzyme Sus scrofa
0.391
-
 bradykinin 37°C, mutant H519L Sus scrofa
0.5
-
 bradykinin 37°C, mutant H519K Sus scrofa

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane bound Sus scrofa 16020
-

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ mono-zinc-containing enzyme, lacks any of the typical metal binding motifs found in other zinc metalloproteases Sus scrofa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
91000
-
 x * 91000, wild-type enzyme, SDS-PAGE Sus scrofa

Organism

Organism UniProt Comment Textmining
Sus scrofa
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein
-
 Sus scrofa

Reaction

Reaction Comment Organism Reaction ID
release of any N-terminal amino acid, including proline, that is linked to proline, even from a dipeptide or tripeptide acive site configuration, modeling, Asp449, Asp460, His523, Glu554, and Glu568 are in volved in metal binding in the active site, His429 and His523 are involved in shuttling protons during catalysis Sus scrofa
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0375
-
 recombinant wild-type enzyme, substrate bradykinin Sus scrofa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
bradykinin + H2O hydrolysis of the N-terminal Arg1-Pro2 bond Sus scrofa Arg + des-Arg-bradykinin
-
 ?
Gly-Pro-hydroxyPro + H2O
-
 Sus scrofa Gly + Pro-hydroxyPro
-
 ?

Subunits

Subunits Comment Organism
? x * 91000, wild-type enzyme, SDS-PAGE Sus scrofa

Synonyms

Synonyms Comment Organism
aminopeptidase P
-
 Sus scrofa
AP-P
-
 Sus scrofa
More the enzyme belongs to the peptidase family M24 Sus scrofa
X-Pro aminopeptidase
-
 Sus scrofa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Sus scrofa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.17
-
 bradykinin 37°C, mutant H519K Sus scrofa
1.18
-
 bradykinin 37°C, wild-type enzyme Sus scrofa
1.82
-
 bradykinin 37°C, mutant H519L Sus scrofa
2.94
-
 bradykinin 37°C, mutant H519L Sus scrofa
6.08
-
 bradykinin 37°C, wild-type enzyme Sus scrofa