What is TEV sequence?

What is TEV sequence?

TEV protease (EC 3.4. 22.44, Tobacco Etch Virus nuclear-inclusion-a endopeptidase) is a highly sequence-specific cysteine protease from Tobacco Etch Virus (TEV). Due to its high sequence specificity it is frequently used for the controlled cleavage of fusion proteins in vitro and in vivo.

What kind of protease is TEV?

cysteine protease
The TEV protease is cysteine protease from Tobacco Etch Virus (TEV) which is highly used for the cleavage of fusion proteins and removal of tags from recombinant proteins in vitro or in vivo. This enzyme belongs to chymotrypsin-like proteases and shows high sequence specificity.

How do you use TEV protease?

How Much TEV Protease to Use? Rule of thumb: Use 1 µg TEV protease per 25 µg to 100 µg of substrate (minimum enzyme concentration: 1 unit/mL). Use more enzyme, if the cleavage site of the substrate is occluded sterically or the substrate is aggregated.

Does TEV protease require EDTA?

The “standard” reaction buffer for TEV protease is 50 mM Tris-HCl (pH 8.0), 0.5 mM EDTA and 1mM DTT. The duration of the cleavage reaction is typically overnight, although lots of cleavage will happen in the first few hours and prolonged incubation times may not lead to proportional increases in cleavage.

How does TEV protease recognize the amino acid sequence?

TEV Protease is a highly specific cysteine protease that recognizes the amino-acid sequence Glu-Asn-Leu-Tyr-Phe-Gln- (Gly/Ser) and cleaves between the Gln and Gly/Ser residues. Removal of affinity purification tags such as maltose-binding protein (MBP) or poly-histidine from fusion proteins Optimal activity and stability for up to 24 months

Which is the cut site of TEV protease?

The consensus for these native cut sites is ENLYFQ\\S where ‘\\’ denotes the cleaved peptide bond. Residues of the substrate are labelled P6 to P1 before the cut site and P1’ after the cut site. Early works also measured cleavage of an array of similar substrates to characterise how specific the protease was for the native sequence.

What is the protease sequence of tobacco etch virus?

Bulk packaging may also be available and requested for large recurring orders. Product Information TEV Protease, also known as Tobacco Etch Virus (TEV) Protease, is a highly specific cysteine protease that recognizes the amino-acid sequence Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser) and cleaves between the Gln and Gly/Ser residues.

Can a directed evolution change TEV protease specificity?

Although rational design has had limited success in changing protease specificity, directed evolution has been used to change the preferred residue either before or after the cleavage site. However, TEV protease does have limitations as a biochemical tool.