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Literature summary for 3.4.21.22 extracted from

  • Yang, L.; Gopalakrishna, K.; Manithody, C.; Rezaie, A.R.
    Expression, purification and characterization of factor IX derivatives using a novel vector system (2006), Protein Expr. Purif., 50, 196-202.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
vitamin K dependent on Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in HEK-293 cells Homo sapiens

Protein Variants

Protein Variants Comment Organism
E186A site-directed mutagenesis, the fIXa mutant exhibits normal activity towards a fIXa-specific chromogenic substrate in the presence of Ca2+ with no obvious requirement for Na+ in the reaction, the mutant interacts with factor VIIIa with near normal affinity and catalyzes the activation of factor X in the intrinsic Xase complex with a normal catalytic efficiency Homo sapiens
H185A site-directed mutagenesis, the fIXa mutant exhibits normal activity towards a fIXa-specific chromogenic substrate in the presence of Ca2+ with no obvious requirement for Na+ in the reaction, the mutant interacts with factor VIIIa with near normal affinity and catalyzes the activation of factor X in the intrinsic Xase complex with a normal catalytic efficiency Homo sapiens
R188A site-directed mutagenesis, the fIXa mutant exhibits normal activity towards a fIXa-specific chromogenic substrate in the presence of Ca2+ with no obvious requirement for Na+ in the reaction, the mutant interacts with factor VIIIa with near normal affinity and catalyzes the activation of factor X in the intrinsic Xase complex with a normal catalytic efficiency Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
EDTA complete inhibition at 20 mM Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information steady-state kinetics of wild-type and mutant enzymes n presence or absence of NaCl Homo sapiens
2.2
-
CH3SO2-D-Leu-Gly-Arg-4-nitroanilide pH 7.5, 37°C, recombinant wild-type enzyme, in absence of NaCl Homo sapiens
2.2
-
CH3SO2-D-Leu-Gly-Arg-4-nitroanilide pH 7.5, 37°C, recombinant wild-type enzyme, in presence of 0.2 M NaCl Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ activates Homo sapiens
Na+ no requirement for Na+ in presence of Ca2+, can partially substite for Ca2+ Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
factor X + H2O Homo sapiens
-
activated factor X + ?
-
?
additional information Homo sapiens the enzyme is a trypsin-like serine protease in plasma that binds to its cofactor factor VIIIa on negatively charged membrane surfaces in the presence of Ca2+, i.e. intrinsic Xase, to activate factor X to factor Xa in the clotting cascade, factor IXa plays an essential role in hemostasis and its deficiency is associated with the life threatening disease, hemophilia B ?
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens P00740
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification proteolytic cleavages at two peptide bonds after Arg145 and Arg180 by either one of two physiological activators, the factor VIIa-tissue factor complex or factor XIa, leads to removal of the activation peptide and conversion of fIX to a fully active enzyme consisting of a light and a heavy chain held together by a single disulfie bond Homo sapiens

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from HEK-293 cells by immunoaffinity and anion exchange chromatogaphy Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
plasma factor IX circulates in plasma as a single chain inactive zymogen Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
CH3SO2-D-Leu-Gly-Arg-4-nitroanilide + H2O amidolytic activity Homo sapiens ?
-
?
factor X + H2O
-
Homo sapiens activated factor X + ?
-
?
factor X + H2O factor IXa acts as part of phosphocholine/phosphoserine vesicles interacting with factor VIIIa in the intrinsic tenase complex Homo sapiens activated factor X + ?
-
?
additional information the enzyme is a trypsin-like serine protease in plasma that binds to its cofactor factor VIIIa on negatively charged membrane surfaces in the presence of Ca2+, i.e. intrinsic Xase, to activate factor X to factor Xa in the clotting cascade, factor IXa plays an essential role in hemostasis and its deficiency is associated with the life threatening disease, hemophilia B Homo sapiens ?
-
?
additional information the loop harboring the S1 specficity site contains the residues His185, Glu186, and Arg188, which, incontrast to other coagulation factor proteases, e.g. unlike thrombin and factor Xa, do not play a functional role in modulating the catalytic function of fIXa in the intrinsic Xase complex, overview Homo sapiens ?
-
?

Subunits

Subunits Comment Organism
More the fully active enzyme consists of a light and a heavy chain held together by a single disulfie bond Homo sapiens

Synonyms

Synonyms Comment Organism
factor IXa
-
Homo sapiens
FIXa
-
Homo sapiens
intrinsic Xase
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
18.5
-
CH3SO2-D-Leu-Gly-Arg-4-nitroanilide pH 7.5, 37°C, recombinant wild-type enzyme, in absence of NaCl Homo sapiens
20.2
-
CH3SO2-D-Leu-Gly-Arg-4-nitroanilide pH 7.5, 37°C, recombinant wild-type enzyme, in presence of 0.2 M NaCl Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Homo sapiens