Human rhinovirus 3C protease (HRV 3C Protease) is a cysteine protease that recognizes the cleavage site of Leu-Glu-Val-Leu-Phe-Gln-Gly-Pro, commonly referred to as the PreScission Site. It cleaves between Gln and Gly. The recombinant form of the HRV3C protease is a restriction grade protease that has robust activity at 4°C and high specific activity. This HRV 3C Protease is a 47 kDa protein with both GST and His tags so it can be removed by either Ni-chelating or Glutathione (GSH) resin. A mutation in the mouse Amelx tri-tyrosyl domain results in impaired secretion of amelogenin and phenocopies human X-linked amelogenesis.

It is recommended to test the HRV3C Protease cleavage with protease-to-target protein ratio of 1:100 (w/w) in the buffer suitable for the target protein at 40°C overnight, with the target protein concentration at or below 1 mg/ml. Under this condition, almost 95% of target protein is cleaved in most cases. The efficiency of cleavage varies due to the sequences around the cleavage site, the conformation and the solubility of the target protein. Including reducing agent such as DTT or TCEP in the buffer helps the cleavage. Due to its high specificity, more protease can be used without non-specific cleavage of target proteins. No non-specific cleavage of control cleavage protein is observed at protease-to-control cleavage protein ratio of 1:10.

Additional Information

Additional Information