Ultrasensitive Measurement of Ca²⁺ Influx into Lipid Vesicles Induced by Protein Aggregates.

ABSTRACT: To quantify and characterize the potentially toxic protein aggregates associated with neurodegenerative diseases, a high-throughput assay based on measuring the extent of aggregate-induced Ca²⁺ entry into individual lipid vesicles has been developed. This approach was implemented by tethering vesicles containing a Ca²⁺ sensitive fluorescent dye to a passivated surface and measuring changes in the fluorescence as a result of membrane disruption using total internal reflection microscopy. Picomolar concentrations of Aβ42 oligomers could be observed to induce Ca²⁺ influx, which could be inhibited by the addition of a naturally occurring chaperone and a nanobody designed to bind to the Aβ peptide. We show that the assay can be used to study aggregates from othe

SUBMITTER: Flagmeier P 

PROVIDER: S-EPMC5615231 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature