Description:
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TEV protease is a catalytic part of the Nuclear Inclusion protein "a" (NIa) from tobacco etch virus (TEV) (1).
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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.
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Resistant to many widly used protein inhibitors like: PMSF, AEBSF, TLCK, E-64, “Complete” protease inhibitor cocktail (Roche).
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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).
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Active at 4 to 30°C (the enzyme is 3 times less active at 4°C than at 30°C)(6).
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Sensitive to some detergents (7).
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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.
References:
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Nayak, S. et al. (2003) Focus in press.
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Dougherty, W.G. et al. (1989) Virology 172, 302.
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Dougherty, W.G., and Parks, T.D. (1989) Virology 172, 145.
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Dougherty, W.G. et al. (1988) EMBO 7,1281.
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Kapust, R.B., at al. (2002a) Biochem.Biophys. Res. Commun.294: 949-955.
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Nallamsrtty, S. at al. (2004) Protein Expr Purif. 38(1): 108-15.
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Mohanty, A.K. at al. (2003) Protein Expr Purif. 27: 109-114.