SARS-CoV 3CL Protease

Product Information

Product Data
Storage/Stability
References

Product Description

Generally, viruses have proteases to process their proteins into active form. Because of its pivotal role in the viral life cycle, proteases are primary targets for the development of antiviral agents. 3CL protease, a viral cysteine proteinase, plays an important role in co-translational proteolytic processing of coronavirus polyproteins. The 3CL protease cleaves as much as 11 sites in the replicase polyproteins and also releases the key replicative functions of polymerase and helicase. 3CL protease is the only coronavirus protein for which structural information is available. 3CL protease comprises three domains, the substrate-binding site is expected to be located between domains I and II, and domain III is a globular cluster comprising five helices. 3CL protease is a homodimer.

Sequence
1 SGFRKMAFPS GKVEGCMVQV TCGTTTLNGL WLDDTVYCPR HVICTAEDML
51 NPNYEDLLIR KSNHSFLVQA GNVQLRVIGH SMQNCLLRLK VDTSNPKTPK
101 YKFVRIQPGQ TFSVLACYNG SPSGVYQCAM RPNHTIKGSF LNGSCGSVGF
151 NIDYDCVSFC YMHHMELPTG VHAGTDLEGK FYGPFVDRQT AQAAGTDTTI
201 TLNVLAWLYA AVINGDRWFL NRFTTTLNDF NLVAMKYNYE PLTQDHVDIL
251 GPLSAQTGIA VLDMCAALKE LLQNGMNGRT ILGSTILEDE FTPFDVVRQC
301 SGVTFQ

Product Data

Purified SARS-3CL Protease protein was run on a 15% SDS-acrylamide gel and stained by Coomassie Blue. Arrow highlights the 3CL protease band.

Lane M: Molecular weight markers
Lanes 1-3: 2 µg, 4 µg, and 6 µg of the protein, respectively.

Click for gel image (opens in a new browser window).

Storage/Stability

Store at -80°C.  Stable for one year under these conditions. Avoid repeated freeze-thaw cycles.

References

Zuo, X., et al. (2005). “Expression and purification of SARS Coronavirus Proteins using SUMO Fusions.” Protein Expres Purif (In Press).

Zuo, Xun., et al. (2005). “Enhanced expression and purification of membrane proteins by SUMO fusion in Escherichia coli.” J Struct Funct Genomics (In Press).

Malakhov, M. P., et al. (2004). “SUMO fusion and SUMO-specific protease for efficient expression and purification of proteins.” J Struct Funct Genomics 5(1-2): 75-86.

Butt, T. R. et al. (2005). “SUMO fusion technology for difficult-to-express proteins.” Protein Expres Purif (In Press).

Marra, M.A., et al. (2003). "The genome sequence of the SARS-associated coronavirus." Science 300(5624): 1399-1404.

Sun H. et al. (2003). "Molecular cloning, expression, purification, and mass spectrometric characterization of 3C-like protease of SARS coronavirus." Protein Expres Purif 32(2): 302-308.

X. Xu, et al. (2003). "Molecular model of SARS coronavirus polymerase: implications for biochemical functions and drug design." Nucleic Acids Res. 31(24): 7117-7130.

T.G. Ksiazek, et. al. (2003). "A novel coronavirus associated with severe acute respiratory syndrome." N. Engl. J. Med. 348(20): 1953-1966.

P.A. Rota, et al. (2003). "Characterization of a novel coronavirus associated with severe acute respiratory syndrome." Science 300(5624): 1394-1399.

S. Muller, et al. (2001). "SUMO, ubiquitin's mysterious cousin." Nat. Rev. Mol. Cell Biol.2(3) 202-210.

F. Melchior (2000). "SUMO--nonclassical ubiquitin." Annu. Rev. Cell Dev. Biol. 16: 591-626.