Difference between revisions of "Part:BBa K3989002"
 
(19 intermediate revisions by the same user not shown)
Line 3: Line 3:
 
<partinfo>BBa_K3989002 short</partinfo>
 
<partinfo>BBa_K3989002 short</partinfo>
  
−
eTEV protease is mutant from the original TEV protease. Seven amino acids(S3I, P8Q, S31T, E79G, T173A, V219R, A231V) are mutated in this variant and the catalytic efficiency is 2-fold higher than the original one.
+
eTEV protease is a variant from the original TEV protease. Seven amino acids(S3I, P8Q, S31T, E79G, T173A, V219R, A231V) are mutated in this variant and the catalytic efficiency is higher than the original one.
 +
 
  
−
<!-- Add more about the biology of this part here
 
 
===Structure and binding site===
 
===Structure and binding site===
 +
<html>
 +
This variant is generated via a directed evaluation system called <b>YESS 2.0</b> (Yeast Endoplasmic Reticulum (ER) Sequestration Screening). The structure is shown below.(Fig.A) Particularly, the sub-structure of at the binding site is shown as well.(Fig. B) The blue amino acid residues represent the changing sites of the variant and the purple residue represent the binding and cutting site of the ligand.
 +
<figure>
 +
  <img src="https://parts.igem.org/wiki/images/b/bb/21_UZurich_eTEV_structure.jpeg">
 +
  <figcaption><b>Fig. A </b> Structural model of eTEV </figcaption>
 +
</figure>
  
  
 +
<figure>
 +
  <img src="https://static.igem.org/mediawiki/parts/2/2b/21_UZurich_eTEV_binding_site.jpeg">
 +
  <figcaption><b>Fig. B </b> Sub-structure at the binding site of eTEV. </figcaption>
 +
</figure>
 +
</html>
  
−
<!-- -->
+
===Performance===
−
<span class='h3bb'>Sequence and Features</span>
+
<html>
−
<partinfo>BBa_K3989002 SequenceAndFeatures</partinfo>
+
Besides this variant, the author also reported some other variants(for instance, <a href="https://parts.igem.org/Part:BBa_K3989001"> BBa_K3989001</a>) with different mutations. Different mutations in the TEV protease ligand-binding site show different catalytic efficincy. The results are shown in Fig. C.
  
 +
<figure>
 +
  <img src="https://static.igem.org/mediawiki/parts/0/09/21_UZurich_catalytic_efficiency.jpeg">
 +
  <figcaption><b>Fig. C </b> Catalytic efficiencies of different TEV protease variants. </figcaption>
 +
</figure>
 +
</html>
  
−
<!-- Uncomment this to enable Functional Parameter display
+
===Sequence and Features===
−
===Functional Parameters===
+
<partinfo>BBa_K3989002 SequenceAndFeatures</partinfo>
−
<partinfo>BBa_K3989002 parameters</partinfo>
+
−
<!-- -->
+
  
 
===Reference===
 
===Reference===

Latest revision as of 10:45, 19 October 2021


eTEV, efficiency-enhanced TEV protease variant

eTEV protease is a variant from the original TEV protease. Seven amino acids(S3I, P8Q, S31T, E79G, T173A, V219R, A231V) are mutated in this variant and the catalytic efficiency is higher than the original one.


Structure and binding site

This variant is generated via a directed evaluation system called YESS 2.0 (Yeast Endoplasmic Reticulum (ER) Sequestration Screening). The structure is shown below.(Fig.A) Particularly, the sub-structure of at the binding site is shown as well.(Fig. B) The blue amino acid residues represent the changing sites of the variant and the purple residue represent the binding and cutting site of the ligand.

Fig. A Structural model of eTEV
Fig. B Sub-structure at the binding site of eTEV.

Performance

Besides this variant, the author also reported some other variants(for instance, BBa_K3989001) with different mutations. Different mutations in the TEV protease ligand-binding site show different catalytic efficincy. The results are shown in Fig. C.

Fig. C Catalytic efficiencies of different TEV protease variants.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Reference

1) Denard, C. A., Paresi, C., Yaghi, R., McGinnis, N., Bennett, Z., Yi, L., ... & Iverson, B. L. (2021). YESS 2.0, a Tunable Platform for Enzyme Evolution, Yields Highly Active TEV Protease Variants. ACS Synthetic Biology, 10(1), 63-71.