Flagmeier, P;
De, S;
Wirthensohn, DC;
Lee, SF;
Vincke, C;
Muyldermans, S;
Knowles, TPJ;
... Klenerman, D; + view all
(2017)
Ultrasensitive Measurement of Ca2+ Influx into Lipid Vesicles Induced by Protein Aggregates.
Angewandte Chemie - International Edition
, 56
(27)
pp. 7750-7754.
10.1002/anie.201700966.
Preview |
Text (Published article)
Flagmeier_Ultrasensitive_Measurement_Ca2+_VoR.pdf - Published Version Download (2MB) | Preview |
Preview |
Text (Supplementary information)
Flagmeier_Ultrasensitive_Measurement_Ca2+_Suppl.pdf Download (3MB) | Preview |
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 other proteins, such as α-synuclein, and to probe the effects of complex biofluids, such as cerebrospinal fluid, and thus has wide applicability.
| Type: | Article |
|---|---|
| Title: | Ultrasensitive Measurement of Ca2+ Influx into Lipid Vesicles Induced by Protein Aggregates |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1002/anie.201700966 |
| Publisher version: | http://doi.org/10.1002/anie.201700966 |
| Language: | English |
| Additional information: | Copyright © 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
| Keywords: | Alzheimer's disease; fluorescence imaging; nano-scale biophysics; neurodegeneration; protein aggregation |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Clinical and Movement Neurosciences |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1558219 |
Archive Staff Only
 |
View Item |
