The invention relates to chimeric proteins comprising an artificial sequence for cleavage by thrombin, in which the activation peptide is fibrinopeptide A. Preferably, said chimeric proteins are derived from the zymogen of a serine protease such as PC or FX, by replacing the activation peptide of...
Addition of protein C or TAFI to cultured endothelial cells in the presence of thrombin results in cleavage of these substrates in a TM-dependent fashion55–57. Several previous reports have used cultured endothelial cells to assess the effects of various proteins (e.g., the platelet α-granule...
Introduction of thrombin sensitive cleavage sequences into the activation peptide of Factor X was accomplished using the four protein engineering strategies described below. It is known that two N-glycans located on amino acids 181 and 191 of wild type Factor X (SEQ ID NO: 1) are important for...
The specific cleavage of a peptide bond of a natural substrate occurs at the carbonyl Scientific REPOrts | 7: 14862 | DOI:10.1038/s41598-017-13868-1 4 www.nature.com/scientificreports/ Figure 6. MD trajectories of JJ1 in complex with thrombin. (A) Time evolutions of the center of ...
Alternatively, the end-point can also refer to the time that is necessary to reach a steady state of the cleavage reaction by thrombin in a normal sample. By washing off non-immobilised, thrombin-cleaved chelate-containing substrate fragments and non-immobilised, non-thrombin-cleaved substrate, th...
A novel purification method for histidine-tagged proteins containing a thrombin cleavage site. Anal. Biochem. 295:180-185.Hefti MH, Van Vugt-Van der Toorn CJG, Dixon R, Vervoort J. A novel purification method for histidine-tagged proteins containing a thrombin cleavage site. Anal. Biochem. ...
However, SdrG does not bind to thrombin-digested Fg. Thus, SdrG appears to target the thrombin cleavage site in the Fg Bβ chain. In fact, SdrG was found to inhibit thrombin-induced fibrinogen clotting by interfering with fibrinopeptide B release....
After 15 min at 37°C, the chromogenic substrate was added and the measurement of the cleavage reaction immediately started in a plate reader at 405 nm. All measurements were performed in duplicates. Model Generation The model of the different inhibitors on thrombin was generated on a Silicon ...
Mutations of ECL2 aspartic acid residues D224A, D230A/D235A (double mutation), and D22A/D230A/D235A mutants were all found to decrease thrombin cleavage of PAR4 and activation in yeast models (57). Furthermore, the activity of the PAR4 agonist peptide AYPGKF-NH2 was also abolished ...