title: Differential temperature dependence of tobacco etch virus and rhinovirus 3C proteases.
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).
authors: Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
authors: Sreejith
authors: S
authors: Raran-Kurussi
authors: József
authors: J
authors: Tözsér
authors: Scott
authors: S
authors: Cherry
authors: Joseph E
authors: JE
authors: Tropea
authors: David S
authors: DS
authors: Waugh
mesh_terms: D000086042:3C Viral Proteases; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012229:Rhinovirus / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D010450:Endopeptidases; C099488:TEV protease; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases
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