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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.
    View publication on PubMed

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