Towards designer organelles by subverting the peroxisomal import pathway

Abstract

The development of ‘designer’ organelles could be a key strategy to enable foreign pathways to be efficiently controlled within eukaryotic biotechnology. A fundamental component of any such system will be the implementation of a bespoke protein import pathway that can selectively deliver constituent proteins to the new compartment in the presence of existing endogenous trafficking systems. Here we show that the protein–protein interactions that control the peroxisomal protein import pathway can be manipulated to create a pair of interacting partners that still support protein import in moss cells, but are orthogonal to the naturally occurring pathways. In addition to providing a valuable experimental tool to give new insights into peroxisomal protein import, the variant receptor-signal sequence pair forms the basis of a system in which normal peroxisomal function is downregulated and replaced with an alternative pathway, an essential first step in the creation of a designer organelle.

Prokaryotic nanocompartments form synthetic organelles in a eukaryote

“…Recent reports have explored the localization of engineered proteins to eukaryotic compartments such as the peroxisome 21,22 , mitochondria 19 , and vacuoles 30 . While peer-reviewed) is the author/funder.…”

“…In contrast to approaches that leverage existing organelles [19][20][21][22] , encapsulins have the advantage of being completely orthogonal to endogenous eukaryotic compartments. There is also ample choice of different encapsulin protein variants derived from different families of bacteria and archaea.…”

Engineering of Metabolic Pathways Using Synthetic Enzyme Complexes

“…Modification of the existing peroxisomal protein import system can increase its efficiency for importing enzymes (DeLoache et al, 2016). However, the recent creation of a different protein import system that runs in parallel with the endogenous system provides a step toward synthetic peroxisomes (Cross et al, 2017). Deeper understanding of proliferation mechanisms and the protein-protein interactions that hold the peroxisomal matrix together will also assist in reaching this goal.…”

Plant peroxisomes at the crossroad of NO and H2O2 metabolism

“…In another case, it has been reported that protein denominated TGBp1 (triple gene block protein 1) from the pepino mosaic virus can interact with catalase to increase its activity and trigger virus accumulation (Mathioudakis et al ). Moreover, the peroxisomal import system has started to be used as a strategy model to generate designed peroxisomes with a protein composition on demand (Cross et al ).…”