Jessica Anderson, John Schultz, Jami English, Jim Hartnett, Christine Andrews, Monika Wood and Bob Bulleit
Promega Corporation, Madison, WI USA
Tobacco Etch Virus Nia Protease (TEV protease) is initially synthesized as a part of a large precursor protein and is processed to a 50kDa form in the plant cell. The published sequence for native cleavage sites for the protein has been identified as EXXYXQ(G/S), with cleavage occurring after the glutamine residue.
In this report we examine the properties of the 50kDa form of the protein (ProTEV Protease) as a protease for cleavage of recombinant fusion proteins, both as purified protein species and in crude cell lysates. The 50kDa form of the protease displays a relatively broad pH optima from pH 6.5–pH 8.5 for most substrates, though some substrates display shifted pH optima. Even though somewhat larger than the typically used form of the protease, ProTEV Protease can effectively digest proteins immobilized on affinity purification resins, releasing the cleaved protein for further studies.