What does factor Xa cleave?
What does factor Xa cleave?
Factor Xa cleaves after the arginine residue in its preferred cleavage site Ile-Glu/Asp-Gly-Arg. It will sometimes cleave at other basic residues, depending on the conformation of the protein substrate. The most common secondary site, among those that have been sequenced, is Gly-Arg.
Where does factor Xa cleave?
Ile-(Glu or Asp)-
Factor Xa cleaves after the arginine residue in its preferred cleavage site Ile-(Glu or Asp)-Gly-Arg. It will sometimes cleave at other basic residues, depending on the conformation of the protein substrate (1,2,3).
Is factor Xa a protease?
Factor X, also known by the eponym Stuart–Prower factor, is an enzyme (EC 3.4. 21.6) of the coagulation cascade. It is a serine endopeptidase (protease group S1, PA clan). Factor X is synthesized in the liver and requires vitamin K for its synthesis.
How does PreScission protease work?
PreScission Protease is a fusion protein of glutathione S-transferase (GST) and human rhinovirus (HRV) type 14 3C protease . The protease specifically recognizes a subset of sequences which include the core amino acid sequence Leu-Phe-Gln/Gly-Pro cleaving between the Gln and Gly residues .
What is factor Xa and thrombin?
Factor Xa, produced by both the extrinsic tenase and intrinsic tenase complexes, represents the point of convergence of the extrinsic and intrinsic pathways of the coagulation cascade and converts prothrombin into thrombin (Figure 1)—the serine protease responsible for fibrin formation and the subsequent growth and …
What is factor Xa in the body?
Factor Xa is a serine protease which cleaves prothrombin to generate thrombin and lies at the crossroads of the extrinsic and intrinsic coagulation pathway. Only a small amount of factor Xa is needed to generate many molecules of thrombin.
What is the role of serine in factor Xa?
How do you cleave his tags?
His-tag removal from protein using TEV Protease
- Dialyze the protein against 20 mM Tris-HCl, pH 7.5.
- Determine the protein concentration.
- Combine 15 μg of protein and H2O (if necessary) to make a 45 μl total reaction volume.
- Add 5 μl of TEV Protease Reaction Buffer (10X) to make a 50 μl total reaction volume.
How much PreScission do I add to protease?
Add 1 µl (2 units) of PreScission Protease for each 100 µg of fusion protein in the eluate.
What is factor 10a?
What is coagulation factor Xa? Coagulation factor Xa is a protein that reverses the effects of certain anticoagulant medications that are used to treat or prevent blood clots. Reversing anticoagulant medicine is necessary if you have uncontrolled or life-threatening bleeding as a result of how that medicine works.
What is factor Xa responsible for?
What is Argatroban?
Argatroban is used to decrease the clotting ability of the blood and to help prevent harmful clots from forming in the blood vessels. This medicine is used to treat or prevent blood clots in patients with bleeding problems caused by another medicine called heparin.
Does factor Xa cleave proline or arginine?
Factor Xa: Factor Xa cleaves after the arginine residue in its preferred cleavage site Ile- (Glu or Asp)-Gly-Arg. It will not cleave a site followed by a proline or arginine. Factor Xa contains two disulfide-linked polypeptide chains with molecular weights of 17 and 42 kDa.
What is factor Xa protease?
Factor Xa Protease. Factor Xa cleaves after the arginine residue in its preferred cleavage site Ile-Glu/Asp-Gly-Arg. It will sometimes cleave at other basic residues, depending on the conformation of the protein substrate. The most common secondary site, among those that have been sequenced, is Gly-Arg. Compatible for use with the pMAL™ Protein
How do you remove thrombin and Factor Xa from a protein?
Thrombin or Factor Xa is removed by connecting a second (Benzamidine) column. Tagged proteases are removed on the same column as used for capture. An affinity tag can be cleaved from your target protein by first eluting the protein from an affinity resin and then adding a protease.
How much factor Xa does it take to cleave MBP?
1 μg of Factor Xa will cleave 50 μg of MBP fusion protein test substrate, MBP-ΔSal to 95% completion in a total reaction volume of 50 μl in 6 hours or less at 23°C in 20 mM Tris-HCl (pH 8.0 @ 25°C) with 100 mM NaCl and 2 mM CaCl2.
