[2] Due to its high sequence specificity, TEV protease is frequently used for the controlled cleavage of fusion proteins in vitro and in vivo.

[4] The tobacco etch virus encodes its entire genome as a single massive polyprotein (350 kDa).

[5] It is composed of two β-barrels and a flexible C-terminal tail and displays structural homology to the chymotrypsin superfamily of proteases (PA clan, C4 family by MEROPS classification).

Early works also measured cleavage of an array of similar substrates to characterise how specific the protease was for the native sequence.

This tunnel contains a set of tight binding pockets such that each side chain of the substrate peptide (P6 to P1’) is bound in a complementary site (S6 to S1’).

It has also been shown that expression can be improved by fusion to maltose binding protein (MBP) which acts a solubility-enhancing partner.

[19] The original TEV protease required the presence of reducing agent for high activity, which could interfere with the function of proteins containing disulfide bonds.

After incorporation of various mutations, later "superTEV protease" versions are highly active in the presence or absence of reducing agent.