Coding

Part:BBa_K1639008

Designed by: Mustafa Yılmaz   Group: iGEM15_ATOMS-Turkiye   (2015-09-13)
Revision as of 14:51, 13 September 2015 by Mustafa26 (Talk | contribs) (References)

Tev Protease, single S219V mutation in the internal cleavage site

TEV protease (also called Tobacco Etch Virus nuclear inclusion a endopeptidase) is a highly sequence-specific cysteine protease from Tobacco Etch Virus (TEV).The consensus for these native cut sites is ENLYFQ\S where ‘\’ denotes the cleaved peptide bond.One of the main uses of this protein is for removing affinity tags from purified proteins. The reason for the use of TEV protease as a biochemical tool is its high sequence specificity. This specificity allows for the controlled cleavage of proteins when the preference sequence is inserted into flexible loops. It also makes it relatively non-toxic in vivo as the recognized sequence scarcely occurs in proteins[1]. However, TEV protease does have limitations as a biochemical tool. It is prone to deactivation by self-cleavage (autolysis), though this can be abolished through a single S219V mutation in the internal cleavage site.[2]


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2
    Illegal XhoI site found at 744
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 605

References

[1]Parks TD, Leuther KK, Howard ED, Johnston SA, Dougherty WG (February 1994). "Release of proteins and peptides from fusion proteins using a recombinant plant virus proteinase". Anal. Biochem. 216 (2): 413–7. doi:10.1006/abio.1994.1060. PMID 8179197

[2]Kapust RB, Tözsér J, Fox JD, Anderson DE, Cherry S, Copeland TD, Waugh DS (December 2001). "Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency". Protein Eng. 14 (12): 993–1000. doi:10.1093/protein/14.12.993. PMID 11809930


[edit]
Categories
//cds/enzyme/protease
//chassis/prokaryote/ecoli
//collections/probiotics/production
Parameters
n/aTev Protease, single S219V mutation in the internal cleavage site