CleaTag™ PreScission Protease

Overview

Description

CleaTag™ Recombinant PreScission Protease (GST-tagged)

Specificity

PreScission Protease is a fusion protein of human rhinovirus (HRV) 3C protease and GST. HRV 3C Protease is encoded by human rhinovirus 14.
It allows for on-column cleavage of GST tags and protein purification in one step.
 

 

Specific cleavage – between the Gln and Gly residues of the recognition sequence LeuGluValLeuPheGln/GlyPro.

Vectors encoding the HRV 3C protease recognition sequence

pGEX-6P vectors pGEX-6P-1, pGEX-6P-2, and pGEX-6P-3 or other vectors encode the recognition sequence for this 3C protease.

Properties

Form

Clear aqueous solution, filter-sterilized, recombinant human rhinovirus (HRV) 3C protease expressed in HEK293.

Database links

UniProt: P03303 (POLG_HRV14)

Synonyms

PreScission Protease(PSP), HRV 3C protease, 3C protease, PP

Molecular weight

47.8kDa (HRV 3C:~22kDa, GST:26kDa )

Unit definition

One unit is defined as the amount of enzyme needed to cleave 100 μg of fusion protein in 16 hours to 90% completion at 5°C in a buffer containing 50 mM Tris-HCl, pH 7.0, 150 mM NaCl, 1 mM EDTA, and 1 mM DTT.

Applications

Application Image

Highlights

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.

Protocols