Studying the interaction between PEX5 and its full-length cargo proteins in living cells by a novel Forsters resonance energy transfer-based competition assay.

ABSTRACT: The import of the majority of soluble peroxisomal proteins is initiated by the interaction between type-1 peroxisomal targeting signals (PTS1) and their receptor PEX5. PTS1 motifs reside at the extreme C-terminus of proteins and consist of a characteristic tripeptide and a modulatory upstream region. Various PTS1-PEX5 interactions have been studied by biophysical methods using isolated proteins or in heterologous systems such as two-hybrid assays, but a recently established approach based on Försters resonance energy transfer (FRET) allows a quantifying investigation in living cells. FRET is the radiation-free energy transfer between two fluorophores in close proximity and can be used to estimate the fraction of acceptor molecules bound to a donor molecule. For PTS1-PEX5 this method relies

SUBMITTER: Hochreiter B 

PROVIDER: S-EPMC9669585 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature