Stoichiometric analysis of protein complexes by cell fusion and single molecule imaging.

ABSTRACT: The composition, stoichiometry and interactions of supramolecular protein complexes are a critical determinant of biological function. Several techniques have been developed to study molecular interactions and quantify subunit stoichiometry at the single molecule level. However, these typically require artificially low expression levels or detergent isolation to achieve the low fluorophore concentrations required for single molecule imaging, both of which may bias native subunit interactions. Here we present an alternative approach where protein complexes are assembled at physiological concentrations and subsequently diluted in situ for single-molecule level observations while preserving them in a near-native cellular environment. We show that coupling this dilution strategy with fluoresce

SUBMITTER: Singh A 

PROVIDER: S-EPMC7483473 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature