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Literature summary for 3.4.11.1 extracted from

  • Wu, G.P.; Cao, M.J.; Chen, Y.; Liu, B.X.; Su, W.J.
    Leucine aminopeptidase from red sea bream (Pagrus major) skeletal muscle: purification, characterization, cellular location, and tissue distribution (2008), J. Agric. Food Chem., 56, 9653-9660.
    View publication on PubMed
Search for more literature for 3.4.11.1

General Stability

General Stability Organism
the enzyme completely loses its activity upon freezing and thawing Pagrus major

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline 31.1% residual activity at 0.2 mM Pagrus major
Ba2+ slight inhibitory effect at 1 mM Pagrus major
bestatin 22.3% residual activity at 0.01 mM Pagrus major
Ca2+ slight inhibitory effect at 1 mM Pagrus major
Cd2+ at concentration of 1 mM, the enzyme is strongly inactivated by Cd2+ (38.2% residual activity) Pagrus major
Cu2+ at concentration of 1 mM, the enzyme is strongly inactivated by Cu2+ (2.3% residual activity) Pagrus major
E-64 40.4% residual activity at 0.015 mM Pagrus major
EDTA strong inhibitor, the enzyme loses 83% of its activity when it is dialyzed extensively against 1 mM EDTA in 25 mM sodium phosphate buffer (pH 7.0), whereas the loss can be restored by Zn2+ to 97% at 0.05 mM and to 81% at 0.1 mM of its original activity, the recovery of enzymatic activity by Mn2+ reaches 94% at 0.5 mM and 79% at 1 mM, on the other hand, the enzymatic activity recovery by Ca2+ and Mg2+ is lower than that by Zn2+ and Mn2+, the former reaches 46% and the latter, 26%, at 1 mM Pagrus major
EGTA 25.4% residual activity at 5 mM Pagrus major
Fe2+ partially inactivated by Fe2+ (53.9% residual activity at 1 mM) Pagrus major
Mn2+ slight inhibitory effect at 1 mM Pagrus major
pepstatin 83.4% residual activity at 0.15 mM Pagrus major
phenylmethanesulfonyl fluoride 94.94% residual activity at 1 mM Pagrus major
Zn2+ at concentration of 1 mM, the enzyme is strongly inactivated by Zn2+ (35.1% residual activity) Pagrus major

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00155
-
 L-leucyl-7-amido-4-methylcoumarin in 25 mM sodium phosphate buffer (pH 7.0) containing 1 mM dithiothreitol and 0.02% NaN3, at 37°C Pagrus major

Metals/Ions

Metals/Ions Comment Organism Structure
Ba2+ shows slight activation at 0.1 mM Pagrus major
Ca2+ shows slight activation at 0.1 mM Pagrus major
Mg2+ slightly activates the enzymatic activity at concentrations from 0.1 to 1 mM Pagrus major
Mn2+ shows slight activation at 0.1 mM Pagrus major
Zn2+ Zn2+ seem to be its metal cofactor Pagrus major

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
96000
-
 approximately 96000 Da, SDS-PAGE and gel filtration Pagrus major

Organism

Organism UniProt Comment Textmining
Pagrus major
-
 red sea bream
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate fractionation, DEAE-Sephacel column chromatography, Sephacryl S-200 gel filtration, hydroxyapatite column chromatography, and phenyl-Sepharose column chromatography Pagrus major

Source Tissue

Source Tissue Comment Organism Textmining
brain
-
 Pagrus major
-
gill
-
 Pagrus major
-
gut
-
 Pagrus major
-
kidney
-
 Pagrus major
-
liver
-
 Pagrus major
-
pyloric cecum
-
 Pagrus major
-
skeletal muscle
-
 Pagrus major
-
stomach
-
 Pagrus major
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.8
-
 crude cell extract, at 37°C Pagrus major
3977
-
 after 4850.5fold purification, at 37°C Pagrus major

Storage Stability

Storage Stability Organism
-80°C, 10% sucrose, 6 months, more than 90% of the enzyme activity remain Pagrus major

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-alanyl-7-amido-4-methylcoumarin + H2O 20.5% activity compared to L-leucyl-7-amido-4-methylcoumarin Pagrus major L-alanine + 7-amino-4-methylcoumarin
-
 ?
L-arginyl-7-amido-4-methylcoumarin + H2O 70% activity compared to L-leucyl-7-amido-4-methylcoumarin Pagrus major L-arginine + 7-amino-4-methylcoumarin
-
 ?
L-leucyl-7-amido-4-methylcoumarin + H2O preferred substrate Pagrus major L-leucine + 7-amino-4-methylcoumarin
-
 ?
L-tyrosyl-7-amido-4-methylcoumarin + H2O 7.4% activity compared to L-leucyl-7-amido-4-methylcoumarin Pagrus major L-tyrosine + 7-amino-4-methylcoumarin
-
 ?
additional information no activity towards L-prolyl-7-amido-4-methylcoumarin, L-valyl-7-amido-4-methylcoumarin, L-glycyl-7-amido-4-methylcoumarin, tert-butyloxycarbonyl-Phe-Ser-Arg-7-amido-4-methylcoumarin, tert-butyloxycarbonyl-Leu-Arg-Arg-7-amido-4-methylcoumarin, and tert-butyloxycarbonyl-Leu-Lys-Arg-7-amido-4-methylcoumarin Pagrus major ?
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 96000, gel filtration Pagrus major

Synonyms

Synonyms Comment Organism
LAP
-
 Pagrus major
leucine aminopeptidase
-
 Pagrus major

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
-
 Pagrus major

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
10 50 activity decreases above 50°C, the enzyme retains 10% of its optimal activity at 10°C Pagrus major

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30
-
 LAP activity decreases slightly after incubation below 30°C Pagrus major
40
-
 LAP activity decreases rapidly above 40°C Pagrus major

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
17.3
-
 L-arginyl-7-amido-4-methylcoumarin in 25 mM sodium phosphate buffer (pH 7.0) containing 1 mM dithiothreitol and 0.02% NaN3, at 37°C Pagrus major
26.4
-
 L-leucyl-7-amido-4-methylcoumarin in 25 mM sodium phosphate buffer (pH 7.0) containing 1 mM dithiothreitol and 0.02% NaN3, at 37°C Pagrus major

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Pagrus major

pH Range

pH Minimum pH Maximum Comment Organism
5.5 8.5 the enzymatic activity is negligible below pH 5.5 and above 8.5 Pagrus major

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 7.5 moderate activity loss after incubation below pH 6.0, but rapid loss after incubation above pH 7.5 for 48 h Pagrus major

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.00144
-
 in 25 mM sodium phosphate buffer (pH 7.0) containing 1 mM dithiothreitol and 0.02% NaN3, at 37°C Pagrus major bestatin