Cleavage Enzymes

HRV 3C Protease – Cleavage Enzyme

Recombinant type 14 3C protease from human rhinovirus (HRV 3C) is a highly purified, recombinant 6XHis-tagged enzyme, which recognizes the cleavage site LeuGluValLeuPheGln↓GlyPro. The small, 22-kDa size of the protease, with optimal activity at 4 ºC, high specificity, and His-tag fusion make HRV 3C protease an ideal choice for rapid removal of fusion tags, even from labile target proteins.



HRV 3C Protease cleaves fusion proteins more efficiently (lanes 2 and 4) compared to cleavage with a competitor’s protease (lanes 3 and 5).

Experimental Methods
Using a 1:100 (w/w) ratio of protease:target protein, 500 μg of purified Nus•Tag™ enolase fusion protein was incubated in parallel 500 μL reactions at 4°C. The reactions was quenched by adding equal volume 4X SDS Sample Buffer and then immediately placing the samples into a water bath at 75 °C for 5 min. Lane M - PerfectProtein Markers, 10-225 kDa, lane 1 - 3 μg purified Nus•Tag enolase fusion protein, lane 2 - 3 μg Nus•Tag enolase fusion protein with 30-min HRV3C protease reaction, lane 3 - 3 μg Nus•Tag enolase fusion protein with 30-min competitor’s protease reaction, lane 4 - 3 μg Nus•Tag enolase fusion protein with 60-min HRV3C protease reaction, lane 5 - 3 μg Nus•Tag enolase fusion protein with 60-min competitor’s protease reaction.