- TEV protease is a catalytic part of the Nuclear Inclusion protein “a” (NIa) from tobacco etch virus (TEV) (1).
- TEV is a cysteine protease that specifically recognizes and cleaves a linear epitope with general sequence E-X-X-Y-X-Q-(G/S)(2-5) (where X is any amino acid). The cleavage occurs between Q and G/S.The most common sequence is ENLYFQG or ENLYFQS.
- Resistant to many widly used protein inhibitors like: PMSF, AEBSF, TLCK, E-64, “Complete” protease inhibitor cocktail (Roche).
- Robust enzyme active in the wide range of different buffers (with NaCl varied from 0 to 0.4 M and in pH from 4 to 9, enzyme tolerates MES, acetate, phosphate, glycerol and sorbitol).
- Active at 4 to 30°C (the enzyme is 3 times less active at 4°C than at 30°C)(6).
- Sensitive to some detergents (7).
- Extremely useful for removing affinity tags from fusion proteins in conditions friendly for target protein.
Unit definition: One unit is the amount of enzyme required to cleave >85% of 3 µg of control substrate (35 kDa fusion protein) in 1 hour at 30oC.
1 x Reaction Buffer: 25 mM Tris-HCl (pH 8.0), 150 mM NaCl, 14 mM β-mercaptoethanol.
Storage Buffer: 0.4 M NaCl, 50 mM Tris-HCl (pH 7.5), 2 mM EDTA, 1 mM DTT, 50% glycerol.
Storage Conditions: Store at –20°C. Protein is stable at least for 9 months.
Quality Control: Protease is greater than 95% single-band pure without non- specific protease contamination.
1. Nayak, S. et al. (2003) Focus in press.
2. Dougherty, W.G. et al. (1989) Virology 172, 302.
3. Dougherty, W.G., and Parks, T.D. (1989) Virology 172, 145.
4. Dougherty, W.G. et al. (1988) EMBO 7,1281.
5. Kapust, R.B., at al. (2002a) Biochem.Biophys. Res. Commun.294: 949-955.
6. Nallamsrtty, S. at al. (2004) Protein Expr Purif. 38(1): 108-15.
7. Mohanty, A.K. at al. (2003) Protein Expr Purif. 27: 109-114.