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Differential temperature dependence of tobacco etch virus and rhinovirus 3C proteases - PubMed

️Tue Jan 01 2013

Differential temperature dependence of tobacco etch virus and rhinovirus 3C proteases

Sreejith Raran-Kurussi et al. Anal Biochem. 2013.

Abstract

Because of their stringent sequence specificity, the 3C-like proteases from tobacco etch virus (TEV) and human rhinovirus are often used for the removal of affinity tags. The latter enzyme is rumored to have greater catalytic activity at 4 °C, the temperature at which fusion protein substrates are usually digested. Here we report that experiments with fusion protein and peptide substrates confirm this conjecture. Whereas the catalytic efficiency of rhinovirus 3C protease is approximately the same at its optimum temperature (30 °C) and at 4 °C, TEV protease is 10-fold less active at the latter temperature due primarily to a reduction in k(cat).

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Figures

**Figure 1**
Temperature Dependence of Initial Reaction Velocity. MBP-NusG fusion protein substrates were digested by TEV [A] or R3C [B] proteases at the indicated temperatures. In the respective insets (right), representative SDS-PAGE gels are shown for uncleaved (Lane 1) and cleaved (Lanes 2-10; 2, 5, 10, 20, 30, 40, 50, 60, and 120 min, respectively) substrates at 4°C.

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Differential temperature dependence of tobacco etch virus and rhinovirus 3C proteases - PubMed