BBa_K316012 1 TEV TEV protease S219P autocatalysis resistant variant 2010-10-22T11:00:00Z 2015-05-08T01:11:56Z TEV protease is naturally found in Tobacco Etch Virus genome. This part contains the sequence codon optimized for expression in B.subtilis using mwg ??? eurofins [[www.eurofinsdna.com]] proprietary software. TEV protease S219P autocatalysis resistant variant Introduction : This is the nuclear inclusion protease, endogenous to Tobacco Etch Virus and is used in the late lifecycle to cleave polyprotein precursors. The recognition sequence is ENLYFQG/S [http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6W9V-45PMGK3-9P&_user=217827&_coverDate=02%2F01%2F1994&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_acct=C000011279&_version=1&_urlVersion=0&_userid=217827&md5=e075aad3a349720ad9484095d01a65be&searchtype=a]] between QG or QSDue to it???s stringent sequence specificity, TEV is commonly used to cleave genetically engineered proteins. Uses: TEV proteinase is used to cleave fusion proteins. It is useful due to its high degree of specificity [[http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6W9V-45PMGK3-9P&_user=217827&_coverDate=02%2F01%2F1994&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_acct=C000011279&_version=1&_urlVersion=0&_userid=217827&md5=e075aad3a349720ad9484095d01a65be&searchtype=a]] and potential to be used in vivo or in vitro applications. Auto-inactivation Wild type TEV protease also cleaves itself at Met 218 and Ser 219 [[http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WXR-45R86R6-5X&_user=7635175&_coverDate=06%2F20%2F1995&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_acct=C000011279&_version=1&_urlVersion=0&_userid=7635175&md5=0a1521e783c0bd4126b145f3f6d766d4&searchtype=a]] . This leads to auto-inactivation of the TEV protease and progressive loss of activity of the protein. The rate of inactivation is proportional to the concentration of protease [[http://peds.oxfordjournals.org/content/14/12/993.long ]] More stable Mutants have been produced by single amino acid substitutions S219V (AGC(serine) to GTG(valine) and S219P (AGC(serine) to CCG(proline) Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency '''Table I.''' Kinetic parameters for wild-type and mutant TEV proteases with the peptide substrate TENLYFQSGTRR-NH2. Enzyme Km (mM) kcat (s−1) kcat/Km (mM−1 s−1) Wild-type 0.061 ?? 0.010 0.16 ?? 0.01 2.62 ?? 0.46 S219V* 0.041 ?? 0.010 0.19 ?? 0.01 4.63 ?? 1.16 S219P* 0.066 ?? 0.008 0.09 ?? 0.01 1.36 ?? 0.22 S219P* - virtually imperivious to autocatalysis S219V* - retains same activity as wild type Full article can be seen here [[http://peds.oxfordjournals.org/content/14/12/993.long]] false false _440_ 0 7480 9 Not in stock false The part was produced by nucleotide synthesis by mwg ??? eurofin false IC 2010 Team annotation2100977 1 TEV protease range2100977 1 1 717 BBa_K316012_sequence 1 ttattagttcattaactgtgtggcctctttaaccggttgaaacggttcttccggtttatccatgaacactttgtgacctccccaaagaacggaatctgcattcagtctccagcctgaaacccattgttgagcttcttggttcgtcagtaactccatgaagttcttgggaacactcgtaaagtaattattcgtattggtgaagtttgacgcagagtgtatgcccacaataaacccatcccgtgttgacacaagaggagagccacattgcccatcttttgtctgaatccaatgcttccaaaagatgccatcagagctagggaatgtacagctagtgtctgataccattgaggacatagatttggtctgaaagtttgttgtgactaagcagatccgttcttcacgctgaggttcgcgaaacttgagtttctgaggaaacggtggaaagtctttcggcattcgaatgataatcatgtctcttccatctatcagatgttgctgaagcgtcgttgtgtttttgaccttaaagactccatgaagggattgtaccaacagtgtaccattattgcgcctaaagaggtgtttgttcgtgatgatgaatggcccaaagccaatgccatacaagctagtggtatgaccatcactctcattggtcagatggcaaatagtcgagctaatcggattatagtcacgtggtcctttaaacaacgatcttcccat igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z