How much 3C protease?
A good starting point for optimization is to use 1 μg of HRV-3C protease per 100 μg of target protein for 1 hour at 0-8 °C, or 1 μg of HRV-3C protease per 500 μg at 0-8 °C for 12-24 hours. Temperatures up to 30 °C can be used for faster digestion.
What is HRV3C protease?
Human rhinovirus 3C protease (HRV3C Protease) is a cysteine protease that recognizes the cleavage site of Leu-Glu-Val-Leu-Phe-Gln*Gly-Pro. HRV3C Protease is capable of cleaving small peptides with the sequence of polyprotein processing sites. It cleaves after the glutamine residue.
What is PreScission protease?
PreScission Protease is a genetically engineered fusion protein of human rhinovirus 3C protease and glutothione S trasferase (GST). PreScisson protease specifically cleaves between the Gln and Gly residues of the recognition sequence LeuGluValLeuPheGln/GlyPro.
How does PreScission protease work?
PreScission Protease is a fusion protein of glutathione S-transferase (GST) and human rhinovirus (HRV) type 14 3C protease (1). 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 (2).
Which one of the following enzymes is an example of cysteine protease?
Cysteine proteases, also known as thiol proteases, are hydrolase enzymes that degrade proteins. These proteases share a common catalytic mechanism that involves a nucleophilic cysteine thiol in a catalytic triad or dyad….Cysteine protease.
| Cysteine peptidase, CA clan | |
|---|---|
| SMART | SM00645 |
| PROSITE | PDOC00126 |
| MEROPS | C1 |
| SCOP2 | 1aec / SCOPe / SUPFAM |
How do I remove GST tag from protein?
Thrombin or Factor Xa can be removed from the protein of interest in one step using a HiTrap™ Benzamidine FF (high sub) column in series after the GSTrap™ column. In this process, the cleaved, tagged protein and thrombin or Factor Xa is washed from the GSTrap™ column onto the HiTrap™ Benzamidine FF (high sub) column.
What is thrombin cleavage?
A thrombin cleavage site (e.g., Leu-Val-Pro-Arg-ll-Gly-Ser; where ll denotes the cleavage site) is widely incorporated within the linker region of fusion or affinity tagged recombinant proteins. The kit contains active thrombin enzyme sufficient to cleave up to 5 mg of the target protein.
How do proteases break down proteins?
Proteases are involved in digesting long protein chains into shorter fragments by splitting the peptide bonds that link amino acid residues.