Difference between revisions of "Part:BBa K2271105"
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K2271105 short</partinfo>
 
<partinfo>BBa_K2271105 short</partinfo>
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<h1>PEX5</h1>
 
<h1>PEX5</h1>
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<h2>Brief introduction</h2>
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  <h2>Brief introduction</h2>
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<p>
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    <p>
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PEX5 is one of two proteins that mediate most of the protein import into the peroxisomal matrix. It recognizes the very c-terminal peroxisomal targeting signal 1 (<b>PTS1</b>), then folds itself to a more compact version and interacts with a variety of enzymes.
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      Protein import into the peroxisome is mediated by two so called peroxins &minus; PEX5 and PEX7. PEX5 is the protein that is responsible for most of the protein import into the peroxisomal membrane. It detects the very twelve amino acids at the C-terminus and then mediates the import of the protein attached to it.
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<br>
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      <br>
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The whole process is divided into five steps: binding, transport, docking, translocation and receptor recycling.
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      The PEX5 of <i>Saccharomyces cerevisiae</i> is a 612 amino acid long proteins, that contains seven tetratricopeptide (TPR) regions which are interacting motifs of the receptor.
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</p>
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      <img src="https://static.igem.org/mediawiki/2017/c/c5/Artico_p5wt_polar.png">
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    </p>
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<h2>Variant</h2>
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      <img src="https://static.igem.org/mediawiki/2017/e/e7/Artico_p5shuttle.jpeg">
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<p>
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      <cite>Peroxisomal matrix protein import: the transient pore model, Erdmann et al. (2005)</cite>
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We followed a targeted mutagenesis approach to achieve an orthogonal import mechanism &minus; our PEX5 variant should recognize a non-native PTS1 variant which is not recognized with the wildtype PEX5.
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    <p>
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</p>
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      The figure above shows all steps of the import mechanisms. It starts with the binding of the PTS1, then the transport to the membrane, where PEX5 interacts with PEX13, PEX14 and PEX17 which leads to membrane integration and pore formation of PEX5. Then the interaction with PEX8, which is bound to PEX2, PEX10 and PEX12, causes cargo release into the matrix. Subsequently ubiquitination of PEX5 lead either to receptor recycling or degradation. This depends on the degree of ubiquitination &minus; while mono- or di-ubiquitination cause recycling, polyubiquitination causes degradation.
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    </p>
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  <h2>Targeted mutagenesis</h2>
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    <p>
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      This biobrick contains a variant of the PEX5 gene which was created in the cause of a targeted mutagenesis approach. This variant should interact with a PTS1 variant instead of the PTS1 signal and thereby provide the necessary basis for an artificial compartment due to an orthogonal import mechanism.
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    </p>
  
  

Revision as of 21:25, 28 October 2017


PEX5 variant R15

PEX5

Brief introduction

Protein import into the peroxisome is mediated by two so called peroxins − PEX5 and PEX7. PEX5 is the protein that is responsible for most of the protein import into the peroxisomal membrane. It detects the very twelve amino acids at the C-terminus and then mediates the import of the protein attached to it.
The PEX5 of Saccharomyces cerevisiae is a 612 amino acid long proteins, that contains seven tetratricopeptide (TPR) regions which are interacting motifs of the receptor. <img src="Artico_p5wt_polar.png"> 

     <img src="Artico_p5shuttle.jpeg">
     Peroxisomal matrix protein import: the transient pore model, Erdmann et al. (2005)

The figure above shows all steps of the import mechanisms. It starts with the binding of the PTS1, then the transport to the membrane, where PEX5 interacts with PEX13, PEX14 and PEX17 which leads to membrane integration and pore formation of PEX5. Then the interaction with PEX8, which is bound to PEX2, PEX10 and PEX12, causes cargo release into the matrix. Subsequently ubiquitination of PEX5 lead either to receptor recycling or degradation. This depends on the degree of ubiquitination − while mono- or di-ubiquitination cause recycling, polyubiquitination causes degradation.

Targeted mutagenesis

This biobrick contains a variant of the PEX5 gene which was created in the cause of a targeted mutagenesis approach. This variant should interact with a PTS1 variant instead of the PTS1 signal and thereby provide the necessary basis for an artificial compartment due to an orthogonal import mechanism.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 409
  • 1000
    COMPATIBLE WITH RFC[1000]