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Team:TU Eindhoven/Background/Membrane Anchors
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<h3>Membrane Anchor Protein CPX</h3> | <h3>Membrane Anchor Protein CPX</h3> | ||
| - | <p>OmpX (Outer Membrane Protein X) is a small, monomeric β-barrel protein that is highly expressed and very useful for protein display applications in <i>E. coli</i>. CPX (Circularly Permuted OmpX) was developed as a bacterial display methodology for N- and C-terminal display. It demonstrated to enable rapid screening of very large peptide libraries with high precision and efficiency. OmpX possesses four extracellular loops, with loops 2 and 3 forming a semi rigid β-sheets protruding from the cell surface. The native N- and C-termini were fused together by a GGSG-linker, and the newly formed N- and C-termini reside on the cell surface. This makes insertion of an unnatural amino acid fairly easy as it can be positioned before the N-terminus or after the C-terminus. By doing this, the protein itself does not have to be modified. See <a href=' | + | <p>OmpX (Outer Membrane Protein X) is a small, monomeric β-barrel protein that is highly expressed and very useful for protein display applications in <i>E. coli</i>. CPX (Circularly Permuted OmpX) was developed as a bacterial display methodology for N- and C-terminal display. It demonstrated to enable rapid screening of very large peptide libraries with high precision and efficiency. OmpX possesses four extracellular loops, with loops 2 and 3 forming a semi rigid β-sheets protruding from the cell surface. The native N- and C-termini were fused together by a GGSG-linker, and the newly formed N- and C-termini reside on the cell surface. This makes insertion of an unnatural amino acid fairly easy as it can be positioned before the N-terminus or after the C-terminus. By doing this, the protein itself does not have to be modified. See <a href='Fig1#'>Figure 1</a> for a schematic overview of CPX and OmpX. |
</p> | </p> | ||
<img id='Fig1' src="https://static.igem.org/mediawiki/2014/5/5e/TU_Eindhoven_OmpX_CPX.png" class="image_wrapper image_fr" width="1085"> | <img id='Fig1' src="https://static.igem.org/mediawiki/2014/5/5e/TU_Eindhoven_OmpX_CPX.png" class="image_wrapper image_fr" width="1085"> | ||
Revision as of 05:02, 5 October 2014
Membrane Anchors: OmpX and Penn 2012 Anchor
To be able to click molecules over the entire cell surface, a protein to anchor the desired coating to the cell is needed. This protein has to be modified in such a way that it contains an azide displayed on the outside of the cell, where it can react with a DBCO-conjugate. Check also our SPAAC Reaction Page for detailed information.
Membrane Anchor Protein CPX
OmpX (Outer Membrane Protein X) is a small, monomeric β-barrel protein that is highly expressed and very useful for protein display applications in E. coli. CPX (Circularly Permuted OmpX) was developed as a bacterial display methodology for N- and C-terminal display. It demonstrated to enable rapid screening of very large peptide libraries with high precision and efficiency. OmpX possesses four extracellular loops, with loops 2 and 3 forming a semi rigid β-sheets protruding from the cell surface. The native N- and C-termini were fused together by a GGSG-linker, and the newly formed N- and C-termini reside on the cell surface. This makes insertion of an unnatural amino acid fairly easy as it can be positioned before the N-terminus or after the C-terminus. By doing this, the protein itself does not have to be modified. See Figure 1 for a schematic overview of CPX and OmpX.
Figure 1. Structure of OmpX alongside a topological depiction of OmpX and CPX.
