Recombinant 2019-nCoV PLpro/papain-like protease, part of a large replicase polyprotein 1ab (E1564-Y1882), was expressed in E. coli cells with an N-terminal. The purity of the 2019-nCoV PLpro was determined to be approx. 80% by densitometry with molecular weight 38 kDa.

The 2019-nCoV PLpro DUB activity is detected using a ubiquitin based proluciferin substrate that is processed through coupled enzyme reactions to release a light signal. The light intensity is quantitatively correlated with the enzyme activity.

SignalChem Biotech offers a virally encoded cysteine protease called 2019-nCoV papain-like protease (PLpro), also referred to as SARS-CoV-2 PLpro. The cysteine protease acts on three cleavage locations of the viral polyprotein to discharge mature non-structural proteins (nsp) [1, 2, and 3], a process that is crucial for viral replication.

In addition, PLpro possesses deubiquitinating and deISGylating activities, thus negatively regulating the ubiquitin and ISG15-dependent host immune response during viral infection.

PLpro also participates together with nsp4 in the assembly of virally-induced cytoplasmic double-membrane vesicles necessary for viral replication.

Product specification

Activity

Sample Activity Plot.

Sample Activity Plot. Image Credit: SignalChem Biotech

For specific information on a given lot, users can see the related technical datasheet.

Activity

PLpro inhibition profile in the presence of various concentrations of PR-619, a broad spectrum and reversible DUB inhibitor.

PLpro inhibition profile in the presence of various concentrations of PR-619, a broad spectrum and reversible DUB inhibitor. Image Credit: SignalChem Biotech

Specific activity

Purity

Sample Purity Data.

Sample Purity Data. Image Credit: SignalChem Biotech

For specific data on a given lot, users can see the related technical datasheet.

Storage, stability, and shipping

The product should be stored at −70 °C. For optimal storage, the target should be aliquoted into smaller amounts following centrifugation and can be stored at the proposed temperature. For best performance, multiple freeze/thaw cycles and repeated handling should be avoided.

Molecular weight

38 kDa

References

  1. Wu F. et al: A new coronavirus associated with human respiratory disease in China. Nature 579: 265-269, 2020
  2. Báez-Santos Y. M. et al: The SARS-coronavirus papain-like protease: structure, function and inhibition by designed antiviral compounds. Antiviral Res. 115: 21-38, 2015
  3. Lindner H. A. et al: The papain-like protease from the severe acute respiratory syndrome coronavirus is a deubiquitinating enzyme. J Virol. 79:15199-15208, 2005