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Literature summary for 3.4.11.9 extracted from

  • Chang, H.C.; Kung, C.C.; Chang, T.T.; Jao, S.C.; Hsu, Y.T.; Li, W.S.
    Investigation of the proton relay system operative in human cystosolic aminopeptidase P (2018), PLoS ONE, 13, e0190816 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.11.9

Activating Compound

Activating Compound Comment Organism Structure
aminoguanidine reactivates enzyme mutant R353 Homo sapiens
guanidine hydrochloride strongly reactivates enzyme mutant R353 at 0.1-10 mM, approximately 90% of the activity lost in theR535A hcAMPP mutant is regained in the presence of 30 mM guanidine hydrochloride. It has neither an observable rescue effect on wild-type hcAMPP nor on the Y527F mutant Homo sapiens
methylguanidine reactivates enzyme mutant R353 Homo sapiens
N-ethyl-guanidine reactivates enzyme mutant R353 Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
gene XPNPEP1, recombinant expression of wild-type and mutant cytosolic aminopeptidases P (hcAMPP, residues M1 to H623), with a thrombin-cleavable N-terminal His-tag in Escherichia coli strain Rosetta (DE3) Homo sapiens

Protein Variants

Protein Variants Comment Organism
additional information kinetic analysis of MnCl2 activation of wild-type, Y527F and R535A hcAMPPs Homo sapiens
R535A site-directed mutagenesis, the mutant enzyme shows reduced kcat and Km compared to the wild-type enzyme. The respective levels of activity restoration are determined to be 86%, 31%, 16%, and 10% for guanidine hydrochloride, methylguanidine, aminoguanidine and N-ethyl-guanidine Homo sapiens
Y527F site-directed mutagenesis, the mutant enzyme shows reduced kcat and Km compared to the wild-type enzyme Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
hydrazine inactivates wild-type hcAMPP and R535A mutant enzymes Homo sapiens
NaCl activates the wild-type enzyme at 160 mM, inhibits the enzyme mutant R353A at 160 mM Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten steady-state kinetics. Kinetic parameters for R535A hcAMPP measured in the presence and absence of guanidine hydrochloride, overview Homo sapiens
0.144
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A, with 1.0 mM guanidine hydrochloride Homo sapiens
0.163
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A, with 10 mM guanidine hydrochloride Homo sapiens
0.17
-
 bradykinin pH 7.5, 37°C, recombinant wild-type enzyme Homo sapiens
0.2
-
 bradykinin pH 7.5, 37°C, recombinant mutant Y527F Homo sapiens
0.327
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A, with 0.1 mM guanidine hydrochloride Homo sapiens
0.35
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
 Homo sapiens 5829
-
additional information isoforms of the human enzyme include the cytosolic (hcAMPP) and membrane-bound (hmAMPP) aminopeptidase P, encoded by XPNPEP1 and XPNPEP2 genes, and a third hypothetical isoform thought to be present in mitochondrial and cytosolic localizations Homo sapiens
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ activates, required, Km values for Mn2+ are 0.0022 mM for the wild-type enzyme, 0.004 mM for mutant Y527F, and 0.0018 mM for mutant R535A. Kinetic analysis of MnCl2 activation of wild-type, Y527F and R535A hcAMPPs Homo sapiens
additional information a metalloprotease Homo sapiens
NaCl activates the wild-type enzyme at 160 mM, inhibits the enzyme mutant R353A at 160 mM Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens Q9NQW7
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant cytosolic isozymes from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography and ultrafiltration Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
bradykinin + H2O i.e. Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg Homo sapiens des-Arg-bradykinin + Arg
-
 ?
additional information aminopeptidase P targets Xaa-Proline peptides for cleavage. Roles of active site residues Tyr527 and Arg535, both residues make significant contributions to the catalytic efficiency, overview Homo sapiens ?
-
 ?

Subunits

Subunits Comment Organism
? x * 72000, recombinant enzyme, SDS-PAGE Homo sapiens
More the reported X-ray structure of hcAMPP depicts a homodimer with each subunit containing an N-terminal domain, a middle domain, and a C-terminal catalytic domain, two active site divalent metal ions. Dimerization is mediated by the N-terminal domain and Trp477, and conservation of the key active site residues including Asp415, Asp426, His489, Glu523, Glu537, His395, His485, and His498 Homo sapiens

Synonyms

Synonyms Comment Organism
aminopeptidase P
-
 Homo sapiens
AMPP
-
 Homo sapiens
hcAMPP
-
 Homo sapiens
XPNPEP1
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 thermal stability of wild-type and mutant hcAMPPs, overview Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.9
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A Homo sapiens
1.8
-
 bradykinin pH 7.5, 37°C, recombinant mutant Y527F Homo sapiens
2
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A, with 0.1 mM guanidine hydrochloride Homo sapiens
3.2
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A, with 1.0 mM guanidine hydrochloride Homo sapiens
8.3
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A, with 10 mM guanidine hydrochloride Homo sapiens
11
-
 bradykinin pH 7.5, 37°C, recombinant wild-type enzyme Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.57
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A Homo sapiens
6.12
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A, with 0.1 mM guanidine hydrochloride Homo sapiens
9
-
 bradykinin pH 7.5, 37°C, recombinant mutant Y527F Homo sapiens
22.22
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A, with 1.0 mM guanidine hydrochloride Homo sapiens
50.92
-
 bradykinin pH 7.5, 37°C, recombinant mutant R535A, with 10 mM guanidine hydrochloride Homo sapiens
64.71
-
 bradykinin pH 7.5, 37°C, recombinant wild-type enzyme Homo sapiens