What is HRV 3C?

What is HRV 3C?

Recombinant type 14 3C protease from human rhinovirus (HRV 3C) is a recombinant restriction-grade protease. HRV 3C is a highly purified recombinant 6XHis-fusion protein, that recognizes the same cleavage site as the native enzyme: LeuGluValLeuPheGln↓GlyPro.

What does 3C protease do?

The 3C-like protease (3CLpro) or main protease (Mpro), formally known as C30 endopeptidase or 3-chymotrypsin-like protease, is the main protease found in coronaviruses. It cleaves the coronavirus polyprotein at eleven conserved sites. It is a cysteine protease and a member of the PA clan of proteases.

Where does 3C cleave?

It is used to cleave affinity tags from fusion proteins. The optimum recognition site for this enzyme is the sequence Leu-Glu-Val-Leu-Phe-Gln/Gly-Pro(LEVLFQ/GP) and cleavage occurs between the Gln and Gly-Pro residues.

What is thrombin cleavage site?

A thrombin cleavage site (e.g., Leu-Val-Pro-Arg-ll-Gly-Ser; where ll denotes the cleavage site) is widely incorporated within the linker region of fusion or affinity tagged recombinant proteins. After successful cleavage with thrombin, affinity tags or fused proteins can be separated from the target protein.

What is PreScission protease?

PreScission Protease is a genetically engineered fusion protein of human rhinovirus 3C protease and glutothione S trasferase (GST). PreScisson protease specifically cleaves between the Gln and Gly residues of the recognition sequence LeuGluValLeuPheGln/GlyPro.

What is the function of 3CLpro?

3CLpro is also called the main protease (Mpro) and plays a major role in the viral replication. PLpro and 3CLpro cleaves the PP chain into 16 NSPs and out of the 16 NSPs generated, 11 NSPs are generated by the 3CLpro, making this protease one of the major targets for developing anti-SARS-CoV drugs8,9.

What are poly proteins?

Polyproteins are chains of covalently conjoined smaller proteins that occur in nature as versatile means to organize the proteome of viruses including HIV.

What pH is thrombin?

Thrombin is active in the pH range of 5-10. Catalytic optimum is pH 8.3. Thrombin precipitates at pH 5 or less. Thrombin does not require divalent metal ions or cofactors for activity.

How do you use PreScission protease?

For each ml of washed Glutathione Sepharose bed volume, mix 40 µl (80 units) of PreScission Protease with 960 µl of Cleavage Buffer at 5 °C. Add the PreScission Protease mixture to the fusion protein-bound Glutathione Sepharose and gently resuspend. Incubate at 5 °C for 4 hours.

How do I remove GST tag from protein?

Thrombin or Factor Xa can be removed from the protein of interest in one step using a HiTrap™ Benzamidine FF (high sub) column in series after the GSTrap™ column. In this process, the cleaved, tagged protein and thrombin or Factor Xa is washed from the GSTrap™ column onto the HiTrap™ Benzamidine FF (high sub) column.

Which drug is a protease inhibitor?

Protease inhibitor drug names darunavir ethanolate (Prezista) fosamprenavir calcium (Lexiva) ritonavir (Norvir) saquinavir mesylate (Invirase)

What is 6LU7?

6LU7: The crystal structure of COVID-19 main protease in complex with an inhibitor N3. NCBI. National Center for. Biotechnology Information.

What is a viral polyprotein?

3CD polyprotein is made up of the 3C protease and the viral RNA polymerase (3D) that is responsible for genome replication. The efficiency of 3C protease and 3D polymerase activity is altered depending on the cleavage state.

What is HRV 3C protease?

HRV 3C Protease is a highly purified recombinant protein derived from human Rhinovirus type 14 and expressed in E. coli. HRV 3C Protease cleaves a specific amino acid sequence (LeuGluValLeuPheGln ↓ GlyPro) and can be used to cleave fusion tag sequences from recombinant proteins that contain an HRV 3C protease cleavage site.

What is hrv3c used for?

HRV3C is also active in a variety of commonly used buffers, providing further flexibility in experimental design. For Research Use Only. Not for use in diagnostic procedures. Store at -20°C.

What is a 1x HRV 3C cleavage buffer?

1 unit (U) is defined as the amount of enzyme that cleaves at least 95% of the Cleavage Control Fusion Protein (100 µg in 1X HRV 3C Cleavage Buffer) in 16 hours at 4°C. Please see the product’s Certificate of Analysis for information about storage conditions, product components, and technical specifications.

What is the structure of rhinovirus-14 3C protease?

Structure of human rhinovirus 3C protease reveals a trypsin-like polypeptide fold, RNA-binding site, and means for cleaving precursor polyprotein. The structure of human rhinovirus-14 3C protease (3Cpro) has been determined at 2.3 A resolution and refined to an R factor of 0.22.