Visualizing active enzyme complexes using a photoreactive inhibitor for proximity ligation--application on γ-secretase.

ABSTRACT: Here, we present a highly sensitive method to study protein-protein interactions and subcellular location selectively for active multicomponent enzymes. We apply the method on γ-secretase, the enzyme complex that catalyzes the cleavage of the amyloid precursor protein (APP) to generate amyloid β-peptide (Aβ), the major causative agent in Alzheimer disease (AD). The novel assay is based on proximity ligation, which can be used to study protein interactions in situ with very high sensitivity. In traditional proximity ligation assay (PLA), primary antibody recognition is typically accompanied by oligonucleotide-conjugated secondary antibodies as detection probes. Here, we first performed PLA experiments using antibodies against the γ-secretase components presenilin 1 (PS1), containing the cat

SUBMITTER: Schedin-Weiss S 

PROVIDER: S-EPMC3663845 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature