CoV-2 3CL
Escherichia Coli.
Filtered White lyophilized (freeze-dried) powder.
Protein is >98% pure as determined by SDS-PAGE and HPLC analyses.
Description
Recombinant Coronavirus 2019 3CL Protease having a Mw of approximately 33.8 kDa was purified from E. coli. The CoV-2 3CL contains 306 amino acids and purified by proprietary chromatographic technique.
Product Specs
The SARS CoV-2 3CL protease (C30 Endopeptidase), derived from the human SARS-CoV-2 virus, is a cysteine protease of the C30 type. It is located within the NS3 segment of the viral genome. The SARS CoV-2 3CL protease plays a crucial role in the virus's life cycle by processing viral polyproteins into their functional, mature units. This protease, also known as the main protease, is essential for CoV replication and is classified as non-structural protein 5. SARS CoV-2 3CL cleaves the CoV polyprotein at 11 specific sites. Its active site contains a cysteine-histidine catalytic dyad, which cleaves peptide bonds at a Gln-(Ser/Ala/Gly) motif. Specifically, it targets the bond between glutamine and serine/alanine/glycine at the P1 position. The activity of the SARS CoV-2 3CL protease is indispensable for processing the CoV replicase polyprotein, a critical step in viral replication.
Recombinant Coronavirus 2019 3CL Protease, with an approximate molecular weight of 33.8 kDa, was produced using E. coli and subsequently purified. This protein comprises 306 amino acids and was purified using a proprietary chromatographic method.
White lyophilized (freeze-dried) powder that has been filtered.
The CoV-2 3CL solution is formulated with 5% Trehalose, 0.02% Tween-20, and PBS at a pH of 7.4.
To prepare a working stock solution, it is recommended to add deionized water to the lyophilized pellet, aiming for a concentration of approximately 0.5 mg/ml. Allow sufficient time for the pellet to dissolve completely. Please note that the CoV-2 3CL is not sterile. Before using it in cell culture, it is crucial to filter the product through an appropriate sterile filter.
For long-term storage, keep the lyophilized protein at -20°C. Once reconstituted, aliquot the protein to prevent repeated freeze-thaw cycles, which can degrade the protein. While the reconstituted protein can be stored at 4°C for a limited period, it is important to note that stability beyond two weeks at this temperature is not guaranteed.
The purity of the protein is greater than 98%, as determined by SDS-PAGE and HPLC analyses.
CoV-2 3CL, a cysteine protease, exhibits specific cleavage activity towards proteins containing the amino acid sequence LQ(S/A/G). The cleavage occurs C-terminal to the glutamine residue.
Escherichia Coli.
SGFRKMAFPS GKVEGCMVQV TCGTTTLNGL WLDDVVYCPR HVICTSEDML NPNYEDLLIR KSNHNFLVQA GNVQLRVIGH SMQNCVLKLK VDTANPKTPK YKFVRIQPGQ TFSVLACYNG SPSGVYQCAM RPNFTIKGSF LNGSCGSVGF NIDYDCVSFC YMHHMELPTG VHAGTDLEGN FYGPFVDRQT AQAAGTDTTI TVNVLAWLYA AVINGDRWFL NRFTTTLNDF NLVAMKYNYE PLTQDHVDIL GPLSAQTGIA VLDMCASLKE LLQNGMNGRT ILGSALLEDE FTPFDVVRQC SGVTFQ
Product Science Overview
The Coronavirus 2019 3CL Protease, also known as the main protease (Mpro) or 3C-like protease (3CLpro), is a crucial enzyme for the replication of the SARS-CoV-2 virus, which causes COVID-19 . This protease is responsible for cleaving the viral polyprotein into functional proteins necessary for the virus’s life cycle . The recombinant form of this protease is used extensively in research to develop antiviral drugs and understand the virus’s mechanisms.
The 3CL protease is a cysteine protease that consists of three domains: domain I, domain II, and domain III . Domains I and II form a chymotrypsin-like fold, while domain III is involved in dimerization, which is essential for the protease’s catalytic activity . The active site of the protease is located in a cleft between domains I and II and contains a catalytic dyad composed of cysteine and histidine residues .
The 3CL protease plays a pivotal role in the viral replication process by cleaving the viral polyprotein at specific sites to release non-structural proteins . These non-structural proteins include RNA-dependent RNA polymerase, helicase, and other enzymes essential for viral RNA synthesis and processing . The protease’s activity is crucial for the virus’s ability to replicate and produce new virions.
Recombinant 3CL protease is produced using various expression systems, such as Escherichia coli, to obtain large quantities of the enzyme for research purposes . The recombinant form retains the same structural and functional properties as the native protease, making it an invaluable tool for studying the enzyme’s activity and screening potential inhibitors .
Given its essential role in viral replication, the 3CL protease is a prime target for antiviral drug development . Researchers have identified several classes of inhibitors, including peptidomimetic covalent inhibitors, non-peptidomimetic covalent inhibitors, and non-covalent small molecule inhibitors . These inhibitors bind to the active site of the protease, preventing it from cleaving the viral polyprotein and thereby inhibiting viral replication .
