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Literature summary for 3.4.24.36 extracted from

  • Mazumder, S.; Ganguly, A.; Ali, N.
    The effect of C-terminal domain deletion on the catalytic activity of Leishmania donovani surface proteinase GP63: Role of Ser446 in proteolysis (2010), Biochimie, 92, 1876-1885.
    View publication on PubMed
Search for more literature for 3.4.24.36

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination, analysis and comparison, overexpression in Escherichia coli strain BL21 (DE3) pLysS Leishmania donovani donovani

Protein Variants

Protein Variants Comment Organism
F448V site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Leishmania donovani donovani
F448Y site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Leishmania donovani donovani
additional information construction of several truncation mutants, deletion of about 180-211 amino acids, DELTA420 and DELTA389, from the C-terminal region results in approximately 50% reduction in residual activitycompared to the wild-type enzyme Leishmania donovani donovani
S446A site-directed mutagenesis, the mutant shows 50% reduced activity and altered kinetics compared to the wild-type enzyme Leishmania donovani donovani
S446T site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Leishmania donovani donovani

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0037
-
 casein fluorogenic substrate, pH 7.2, 37°C, wild-type enzyme Leishmania donovani donovani
0.005
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant F448V Leishmania donovani donovani
0.0052
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant F448Y Leishmania donovani donovani
0.0053
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA542 Leishmania donovani donovani
0.0054
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant S446T Leishmania donovani donovani
0.0055
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA484 Leishmania donovani donovani
0.0098
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant S446A Leishmania donovani donovani
0.0104
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA389 Leishmania donovani donovani
0.011
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA420 Leishmania donovani donovani

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ zinc proteinase Leishmania donovani donovani

Organism

Organism UniProt Comment Textmining
Leishmania donovani donovani C7EX18
-
-
Leishmania donovani donovani MHOM/IN/1983/AG83 C7EX18
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography and ultrafiltration Leishmania donovani donovani

Source Tissue

Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
azocasein + H2O
-
 Leishmania donovani donovani ?
-
 ?
azocasein + H2O
-
 Leishmania donovani donovani MHOM/IN/1983/AG83 ?
-
 ?
casein + H2O fluorescein isothiocyanate-labeled casein Leishmania donovani donovani ?
-
 ?
casein + H2O fluorescein isothiocyanate-labeled casein Leishmania donovani donovani MHOM/IN/1983/AG83 ?
-
 ?
Gelatin + H2O
-
 Leishmania donovani donovani ?
-
 ?
Gelatin + H2O
-
 Leishmania donovani donovani MHOM/IN/1983/AG83 ?
-
 ?

Subunits

Subunits Comment Organism
More homology modeling and structure-function relationship of GP63, overview Leishmania donovani donovani

Synonyms

Synonyms Comment Organism
gp63
-
 Leishmania donovani donovani
surface proteinase GP63
-
 Leishmania donovani donovani

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Leishmania donovani donovani

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant S446A Leishmania donovani donovani
3.5
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA389 Leishmania donovani donovani
3.7
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA420 Leishmania donovani donovani
11.1
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA484 Leishmania donovani donovani
11.8
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant F448Y Leishmania donovani donovani
11.8
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA542 Leishmania donovani donovani
11.9
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant S446T Leishmania donovani donovani
12
-
 casein fluorogenic substrate, pH 7.2, 37°C, wild-type enzyme Leishmania donovani donovani
12.7
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant F448V Leishmania donovani donovani

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2 7.5 assay at Leishmania donovani donovani

General Information

General Information Comment Organism
evolution GP63 belongs to the metzincin class, encoded by a series of tandemly linked genes usually in a single chromosome that are highly sequence-conserved among different species of Leishmania Leishmania donovani donovani
additional information homology modeling and structure-function relationship of GP63, overview Leishmania donovani donovani

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
300
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant S446A Leishmania donovani donovani
300
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA389 Leishmania donovani donovani
300
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA420 Leishmania donovani donovani
2000
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA484 Leishmania donovani donovani
2200
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant F448Y Leishmania donovani donovani
2200
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant S446T Leishmania donovani donovani
2200
-
 casein fluorogenic substrate, pH 7.2, 37°C, truncated mutant DELTA542 Leishmania donovani donovani
2500
-
 casein fluorogenic substrate, pH 7.2, 37°C, mutant F448V Leishmania donovani donovani
3200
-
 casein fluorogenic substrate, pH 7.2, 37°C, wild-type enzyme Leishmania donovani donovani