Literature summary extracted from
Chan, C.W.; Chan, M.W.; Liu, M.; Fung, L.; Cole, E.H.; Leibowitz, J.L.; Marsden, P.A.; Clark, D.A.; Levy, G.A.
Kinetic analysis of a unique direct prothrombinase, fgl2, and identification of a serine residue critical for the prothrombinase activity (2002), J. Immunol., 168, 5170-5177.
Activating Compound
EC Number |
Activating Compound |
Comment |
Organism |
Structure |
---|
3.4.21.6 |
Factor Va |
absolutely required for activity as part of the prothrombinase complex |
Mus musculus |
|
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
3.4.21.6 |
gene fgl2, expression of the full sized, His-tagged enzyme in Spodoptera frugiperda Sf9 cells via transfection with baculovirus, expression of truncated enzyme forms and enzyme mutants in CHO-K1 cells |
Mus musculus |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
3.4.21.6 |
additional information |
expression of several differently truncated enzymes reveal that residues 285-335 are critical for prothrombinase activity, a truncated enzyme lacking these residues is catalytically inactive |
Mus musculus |
3.4.21.6 |
S135A |
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme |
Mus musculus |
3.4.21.6 |
S425A |
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme |
Mus musculus |
3.4.21.6 |
S89A |
site-directed mutagenesis, completely inactive mutant |
Mus musculus |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
3.4.21.6 |
4-aminobenzamidine |
90% inhibition of factor Xa and 2% inhibition of prothrombinase complex activity at 1 mM |
Mus musculus |
|
3.4.21.6 |
antithrombin III |
90% inhibition of factor Xa at 0.01 mg/ml in presence of heparin and 5% inhibition of prothrombinase complex activity at 1 U/ml |
Mus musculus |
|
3.4.21.6 |
Aprotinin |
95% inhibition of factor Xa and 2% inhibition of prothrombinase complex activity at 10 U/ml |
Mus musculus |
|
3.4.21.6 |
diisopropylfluorophosphate |
2.5 mM: 10% inhibition of factor Xa and 95% inhibition of prothrombinase complex activity, 10 mM: 90% inhibition of factor Xa and 95% inhibition of prothrombinase complex activity |
Mus musculus |
|
3.4.21.6 |
PMSF |
complete inhibition of factor Xa and 5% inhibition of prothrombinase complex activity at 1 mM |
Mus musculus |
|
3.4.21.6 |
Soybean trypsin inhibitor |
complete inhibition of factor Xa and 5% inhibition of prothrombinase complex activity at 0.01 mg/ml |
Mus musculus |
|
KM Value [mM]
EC Number |
KM Value [mM] |
KM Value Maximum [mM] |
Substrate |
Comment |
Organism |
Structure |
---|
3.4.21.6 |
0.0072 |
- |
prothrombin |
pH 7.4, 37°C, wild-type enzyme in presence of factor Va |
Mus musculus |
|
3.4.21.6 |
0.0083 |
- |
prothrombin |
pH 7.4, 37°C, wild-type enzyme |
Mus musculus |
|
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
3.4.21.6 |
Ca2+ |
absolutely required for activity as part of the prothrombinase complex |
Mus musculus |
|
3.4.21.6 |
phosphatidyl-L-serine |
best in lipid vesicles with up to 25% phosphatidyl-L-serine and at least 75% phosphocholine, absolutely required for activity as part of the prothrombinase complex |
Mus musculus |
|
3.4.21.6 |
phosphocholine |
best in lipid vesicles with up to 25% phosphatidyl-L-serine and at least 75% phosphocholine |
Mus musculus |
|
Molecular Weight [Da]
EC Number |
Molecular Weight [Da] |
Molecular Weight Maximum [Da] |
Comment |
Organism |
---|
3.4.21.6 |
65000 |
- |
x * 65000, recombinant wild-type enzyme, SDS-PAGE |
Mus musculus |
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
3.4.21.6 |
prothrombin + H2O |
Mus musculus |
- |
thrombin + ? |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
3.4.21.6 |
Mus musculus |
- |
gene fgl2 |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
3.4.21.6 |
recombinant His-tagged wild-type enzyme from Sf9 insect cells, to homogeneity |
Mus musculus |
Reaction
EC Number |
Reaction |
Comment |
Organism |
Reaction ID |
---|
3.4.21.6 |
selective cleavage of Arg-/-Thr and then Arg-/-Ile bonds in prothrombin to form thrombin |
scutelarin has similar specificity, Ser89 is critical for enzyme activity |
Mus musculus |
|
Specific Activity [micromol/min/mg]
EC Number |
Specific Activity Minimum [µmol/min/mg] |
Specific Activity Maximum [µmol/min/mg] |
Comment |
Organism |
---|
3.4.21.6 |
additional information |
- |
coagulation activity of different, recombinant enzymes forms in presence of varying mixtures of phospholipids |
Mus musculus |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
3.4.21.6 |
D-hexahydrotyrosyl-L-alanyl-L-arginine-4-nitroanilide + H2O |
synthetic chromogenic substrate |
Mus musculus |
D-hexahydrotyrosyl-L-alanyl-L-arginine + 4-nitroaniline |
- |
? |
|
3.4.21.6 |
prothrombin + H2O |
- |
Mus musculus |
thrombin + ? |
- |
? |
|
3.4.21.6 |
prothrombin + H2O |
the recombinantly expressed enzyme is inactive in Sf9 cells, activity can be restored by addition of phosphatidyl-L-serine, and further enhanced by addition of factor Va and Ca2+ |
Mus musculus |
thrombin + ? |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
3.4.21.6 |
? |
x * 65000, recombinant wild-type enzyme, SDS-PAGE |
Mus musculus |
3.4.21.6 |
More |
factor Xa is part of the prothrombinase complex with Ca2+, phospholipids and factor Va |
Mus musculus |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
3.4.21.6 |
factor Xa |
- |
Mus musculus |
3.4.21.6 |
Fgl2 protein |
- |
Mus musculus |
3.4.21.6 |
More |
the enzyme belongs to the peptidase family S1, i.e. trypsin family |
Mus musculus |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
3.4.21.6 |
37 |
- |
assay at |
Mus musculus |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
3.4.21.6 |
7 |
7.4 |
assay at |
Mus musculus |