Part:BBa_K2271105

PEX5 variant R15

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.

Figure 1: TPR domains of the yeasts PEX5 protein.

Figure 2: Import mechanism

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]