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Anal Biochem. 2013 May 15;436(2):142-4. doi: 10.1016/j.ab.2013.01.031. Epub 2013 Feb 7.

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

Author information

1
Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.

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).

PMID:
23395976
PMCID:
PMC4196241
DOI:
10.1016/j.ab.2013.01.031
[Indexed for MEDLINE]
Free PMC Article

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