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Literature summary for 1.4.3.3 extracted from

  • Murtas, G.; Sacchi, S.; Pollegioni, L.
    Substitution of arginine 120 in human D-amino acid oxidase favors FAD-binding and nuclear mistargeting (2019), Front. Mol. Biosci., 6, 125 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.4.3.3

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Homo sapiens

Protein Variants

Protein Variants Comment Organism
R120E substitution in order to mimic the active nuclear translocation signal, slightly alters protein conformation, thermal stability, and kinetic properties, while the dimeric structure and the ligand-binding properties are unchanged. Mutant shows an increase in cytosolic localization that promotes nuclear targeting, without affecting cell viability Homo sapiens
R120L substitution in order to eliminate the positive charge, slightly alters protein conformation, thermal stability, and kinetic properties, while the dimeric structure and the ligand-binding properties are unchanged Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.6
-
 D-serine wild-type, pH 8.5, 25°C Homo sapiens
0.6
-
 D-serine mutant R120E, pH 8.5, 25°C Homo sapiens
0.7
-
 D-serine mutant R120L, pH 8.5, 25°C Homo sapiens
1.3
-
 D-alanine wild-type, pH 8.5, 25°C Homo sapiens
1.4
-
 D-alanine mutant R120E, pH 8.5, 25°C Homo sapiens
2
-
 D-alanine mutant R120L, pH 8.5, 25°C Homo sapiens
7.5
-
 D-cysteine wild-type, pH 8.5, 25°C Homo sapiens
17.6
-
 D-cysteine mutant R120E, pH 8.5, 25°C Homo sapiens
26.7
-
 D-cysteine mutant R120L, pH 8.5, 25°C Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
peroxisome
-
 Homo sapiens 5777
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
80000
-
 mutants R120E and R120L, gel filtration Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens P14920
-
-

Source Tissue

Source Tissue Comment Organism Textmining

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
80
-
 wild-type, substrate D-alanine, pH 8.5, 25°C Homo sapiens
100
-
 mutants R120E and R120L, substrate D-alanine, pH 8.5, 25°C Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-alanine + H2O + O2
-
 Homo sapiens pyruvate + NH3 + H2O2
-
 ?
D-cysteine + H2O + O2
-
 Homo sapiens 2-oxo-3-sulfanylpropionate + NH3 + H2O2
-
 ?
D-serine + H2O + O2
-
 Homo sapiens 3-hydroxypyruvate + NH3 + H2O2
-
 ?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
46.1
-
 mutant R120E, TM value, circular dichroism method Homo sapiens
47.6
-
 mutant R120L, TM value, circular dichroism method Homo sapiens
52.7
-
 mutant R120E, TM value, protein fluoprescence method Homo sapiens
54.2
-
 mutant R120L, TM value, protein fluoprescence method Homo sapiens
57
-
 wild-type, TM value, circular dichroism method Homo sapiens
57.7
-
 wild-type, TM value, protein fluoprescence method Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3
-
 D-cysteine wild-type, pH 8.5, 25°C Homo sapiens
5.2
-
 D-alanine wild-type, pH 8.5, 25°C Homo sapiens
8.5
-
 D-cysteine mutant R120E, pH 8.5, 25°C Homo sapiens
8.6
-
 D-serine wild-type, pH 8.5, 25°C Homo sapiens
9.9
-
 D-serine mutant R120E, pH 8.5, 25°C Homo sapiens
11.3
-
 D-cysteine mutant R120L, pH 8.5, 25°C Homo sapiens
12.6
-
 D-alanine mutant R120E, pH 8.5, 25°C Homo sapiens
15.6
-
 D-serine mutant R120L, pH 8.5, 25°C Homo sapiens
17.3
-
 D-alanine mutant R120L, pH 8.5, 25°C Homo sapiens

Cofactor

Cofactor Comment Organism Structure
FAD R120E and R120L variants show absorbance maxima at 448, 384, and 279 nm and an Abs279nm/Abs448nm ratio of 10.4 and 11.2, respectively, which are close to the value of 10.6 reported for the wild-type enzyme Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.4
-
 D-cysteine wild-type, pH 8.5, 25°C Homo sapiens
0.4
-
 D-cysteine mutant R120L, pH 8.5, 25°C Homo sapiens
0.5
-
 D-cysteine mutant R120E, pH 8.5, 25°C Homo sapiens
4
-
 D-alanine wild-type, pH 8.5, 25°C Homo sapiens
8.5
-
 D-alanine mutant R120L, pH 8.5, 25°C Homo sapiens
8.9
-
 D-alanine mutant R120E, pH 8.5, 25°C Homo sapiens
14.6
-
 D-serine wild-type, pH 8.5, 25°C Homo sapiens
16.5
-
 D-serine mutant R120E, pH 8.5, 25°C Homo sapiens
22.6
-
 D-serine mutant R120L, pH 8.5, 25°C Homo sapiens