Results
Introduction
While there still is much that can be done to improve our toolbox further, we are nonetheless extremely proud of our achievements. Months of lab work payed off! These are our results.
Sub-projects
MD simulations
The first experiments we performed in the wet lab are the tests of the receptors we modelled via molecular dynamics. As soon as we finished building our constructs, we transformed them into PEX5 knock out yeast cells. The results of this experiments can be seen in the following figure.
The fluorescent signal of mTurquoise was detected in the whole cell of each modelled receptor−peptide combination. This indicates that our receptors were not able to recognize the PTS-variants tagged to mTurquoise and thus we did not see import.
PEX5 variant R19
Our second approach for the modification of the importer PEX5 was our so called receptor R19. Based on published literature we built this receptor by replacing three amino acids within the PEX5 protein sequence of the wild type yeast. The corresponding modified PTS1* is characterized by its -SYY sequence at the very end of the peptide. In the following figure one can see the microscopy image of this import system variant cloned in PEX5 knock-out yeast.
