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Literature summary for 3.4.11.1 extracted from

  • Cappiello, M.; Lazzarotti, A.; Buono, F.; Scaloni, A.; D'ambrosio, C.; Amodeo, P.; Mendez, B.L.; Pelosi, P.; Del Corso, A.; Mura, U.
    New role for leucyl aminopeptidase in glutathione turnover (2004), Biochem. J., 378, 35-44.
    View publication on PubMedView publication on EuropePMC

General Stability

General Stability Organism
Mn2+ is absolutely required for stability during purification Bos taurus
Mn2+ is essential for enzyme stability, in absence of Mn2+ the enzyme is inactivated within 48 h Bos taurus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.42
-
Cys-Gly pH 6.9, 25°C Bos taurus
0.57
-
Cys-Gly pH 7.4, 25°C Bos taurus
0.59
-
Cys-Gly pH 8.3, 25°C Bos taurus
1.3
-
L-Met-Gly pH 8.3, 25°C Bos taurus
1.5
-
Met-Gly pH 7.4, 25°C Bos taurus
1.5
-
Leu-Gly pH 8.3, 25°C Bos taurus
2.3
-
Leu-Gly pH 6.9, 25°C Bos taurus
2.5
-
Leu-Gly pH 7.4, 25°C Bos taurus
2.8
-
L-Met-Gly pH 6.9, 25°C Bos taurus
5
-
Ser-Gly pH 8.3, 25°C Bos taurus
5.2
-
L-Ser-Gly pH 7.4, 25°C Bos taurus

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Bos taurus 5829
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ absolutely dependent on Bos taurus
Mn2+ dependent on Bos taurus
additional information unaffected by Zn2+ after removal of bound metal ions, while Mn2+ can restore activity Bos taurus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
52874
-
6 * 53700, SDS-PAGE, 6 * 52874, mass spectrometry Bos taurus
52874
-
x * 52874, mass spectrometry Bos taurus
53700
-
6 * 53700, SDS-PAGE, 6 * 52874, mass spectrometry Bos taurus
320000
-
gel filtration Bos taurus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
Cys-Gly + H2O Bos taurus the enzyme plays a role in glutathione turnover Cys + Gly
-
?
cysteinylglycine + H2O Bos taurus involved in turnover of glutathione cysteine + glycine
-
?

Organism

Organism UniProt Comment Textmining
Bos taurus P00727
-
-

Purification (Commentary)

Purification (Comment) Organism
1090fold, to homogeneity Bos taurus
native enzyme 1090fold from lens to homogeneity by ion exchange chromatography, ethanol and alkaline precipitation, chloroform/octanol extraction, and gel filtration Bos taurus

Reaction

Reaction Comment Organism Reaction ID
release of an N-terminal amino acid, Xaa-/-Yaa-, in which Xaa is preferably Leu, but may be other amino acids including Pro although not Arg or Lys, and Yaa may be Pro. Amino acid amides and methyl esters are also readily hydrolysed, but rates on arylamides are exceedingly low molecular modelling of enzyme-substrate binding Bos taurus
release of an N-terminal amino acid, Xaa-/-Yaa-, in which Xaa is preferably Leu, but may be other amino acids including Pro although not Arg or Lys, and Yaa may be Pro. Amino acid mides and methyl esters are also readily hydrolysed, but rates on arylamides are exceedingly low substrate-enzyme complex structure, molecular modeling Bos taurus

Source Tissue

Source Tissue Comment Organism Textmining
lens
-
Bos taurus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
10.9
-
purified enzyme Bos taurus

Storage Stability

Storage Stability Organism
-80°C, pure enzyme, 0.2 mM MnCl2, 2 weeks, stable Bos taurus
-80°C, purified enzyme after chloroform/octanol extraction purification step, in presence of 0.2 mM Mn2+, stable for at least 2 weeks Bos taurus
4°C, partially purified enzyme after ion exchange chromatography purification step, stable for at least 6 months Bos taurus
4°C, partially purified enzyme, 6 months, stable, less stable after further purification Bos taurus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Cys-Gly + H2O
-
Bos taurus Cys + Gly
-
?
Cys-Gly + H2O the enzyme plays a role in glutathione turnover Bos taurus Cys + Gly
-
?
cysteinylglycine + H2O involved in turnover of glutathione Bos taurus cysteine + glycine
-
?
L-Met-Gly + H2O
-
Bos taurus L-Met + Gly
-
?
L-Ser-Gly + H2O
-
Bos taurus L-Ser + Gly
-
?
Leu-Gly + H2O
-
Bos taurus Leu + Gly
-
?
Met-Gly + H2O
-
Bos taurus Met + Gly
-
?
additional information molecular modelling of enzyme-substrate conformation, no activity with Asp-Gly Bos taurus ?
-
?
additional information no activity with Ser-Gly and Asp-Gly Bos taurus ?
-
?
Ser-Gly + H2O
-
Bos taurus Ser + Gly
-
?

Subunits

Subunits Comment Organism
? x * 52874, mass spectrometry Bos taurus
hexamer 6 * 53700, SDS-PAGE, 6 * 52874, mass spectrometry Bos taurus
More primary structure analysis and molecular modeling Bos taurus

Synonyms

Synonyms Comment Organism
cysteinyl-glycine hydrolysing activity
-
Bos taurus
LAP
-
Bos taurus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Bos taurus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.6
-
Cys-Gly pH 6.9, 25°C Bos taurus
17
-
Ser-Gly pH 7.4, 25°C Bos taurus
42
-
Ser-Gly pH 8.3, 25°C Bos taurus
56.66
-
Cys-Gly pH 6.9, 25°C Bos taurus
57
-
Cys-Gly pH 6.9, 25°C Bos taurus
67
-
Cys-Gly pH 8.3, 25°C Bos taurus
100
-
Cys-Gly pH 7.4, 25°C Bos taurus
118
-
Cys-Gly pH 8.3, 25°C Bos taurus
118.3
-
Cys-Gly pH 8.3, 25°C Bos taurus
135.8
-
Met-Gly pH 6.9, 25°C Bos taurus
136
-
Met-Gly pH 6.9, 25°C Bos taurus
191.7
-
Leu-Gly pH 6.9, 25°C Bos taurus
192
-
Leu-Gly pH 6.9, 25°C Bos taurus
400
-
Leu-Gly pH 7.4, 25°C Bos taurus
468
-
Met-Gly pH 7.4, 25°C Bos taurus
486.3
-
Met-Gly pH 7.4, 25°C Bos taurus
675
-
Leu-Gly pH 8.3, 25°C Bos taurus
988
-
Met-Gly pH 8.3, 25°C Bos taurus
988.3
-
Met-Gly pH 8.3, 25°C Bos taurus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
additional information
-
modeling of pH-dependency Bos taurus
6.9 8.3 assay at Bos taurus
7.4
-
assay at Bos taurus

pH Range

pH Minimum pH Maximum Comment Organism
additional information
-
pH-dependence of activity with different dipeptide substrates Bos taurus