PreScission Protease for GST-tag removal
PreScission Protease is a fusion protein of human rhinovirus (HRV) 3C protease and GST. It allows for on-column cleavage of GST tags and protein purification in one step.
PreScission Protease for GST-tag removal
PreScission Protease is a fusion protein of human rhinovirus (HRV) 3C protease and GST. It allows for on-column cleavage of GST tags and protein purification in one step.
Overview
- Specific cleavage – between the Gln and Gly residues of the recognition sequence LeuGluValLeuPheGln/GlyPro.
- Time-saving – GST-tagged proteins can be cleaved while still bound to an affinity resin.
- Low incubation temperature – enables low temperature cleavage of fusion proteins containing the HRV 3C protease recognition sequence.
On-column or off-column enzyme cleavage
GST tags and protease can be removed in a single step while fusion proteins are still bound to Glutathione Sepharose 4B, Glutathione Sepharose High Performance, or Glutathione Sepharose 4 Fast Flow affinity resin. Another option is to elute the GST-tagged protein using reduced glutathione, cleave, then pass the sample over the same resin to remove the protease and tags.
Unlike other common proteases, such as thrombin and Factor Xa, this GST-tagged fusion protein can be removed easily through binding to a resin with affinity for GST. Other non-recombinant proteases require an additional purification step with a different resin.
Vectors encoding the HRV 3C protease recognition sequence
pGEX-6P vectors pGEX-6P-1, pGEX-6P-2, and pGEX-6P-3 encode the recognition sequence for this 3C protease.
Successful use in published research
PreScission Protease is used for enzyme cleavage in a variety of research applications, including:
- On-column cleavage and purification of proteins involved in ubiquitination
- Cleavage and purification of GST-tagged proteins from inclusion bodies for study of ion channel complex formation in hepatitis C virus
