Tobacco Etch Virus (TEV) NIa protease is a very useful tool in molecular biology. It recognizes a seven amino acid consensus sequence, Glu-X-X-Tyr-X-Gln/Ser, where X can be various amino acyl residues. Cleavage occurs between the conserved Gln and Ser residues. TEV protease is a Cys protease and is thus inhibited by thiol reagents such as Iodoacetamide. Because of its specificity it is often used to remove tags such as 6His tags after protein purification.
We have established the use of TEV protease for Target Directed Proteolysis (TDP) in vivo. Because of its distinct specificity, TEV protease can be expressed in various cellular compartments without interfering with viability. Polypeptides that are not natural substrates of TEV protease are proteolyzed if they carry the appropriate cleavage site. Therefore, TDP allows both structural and functional mapping of a protein of interest in its native environment in vivo.
There are many potential uses for TDP in vivo for example controlled inactivation of essential proteins and topological studies of integral membrane proteins, among others.
These pages provide information on how to use TEV protease.