Chapter title |
Reconstitution of Synaptic SNAREs into Large Liposomes with Reduced Curvature Stress.
|
---|---|
Chapter number | 8 |
Book title |
Synapse Development
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6688-2_8 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6686-8, 978-1-4939-6688-2
|
Authors |
Javier M. Hernandez |
Editors |
Alexandros Poulopoulos |
Abstract |
Liposomes constitute a convenient biochemical model system to investigate mechanistic aspects of the membrane fusion of synaptic vesicles. The proteins responsible for mediating fusion are the SNAREs that belong to a highly conserved family of transmembrane proteins. Reconstituting SNAREs into liposomes using detergents has become a common approach not only to understand how SNAREs work, but also how fusion is regulated by the vast array of accessory proteins present at the presynapse. However, a concern has been that the high curvature stress of the small liposomes (diameters of ~40 nm) frequently used in many studies renders them prone to spontaneous fusion, bringing into question whether the measurements obtained faithfully represent SNARE-mediated fusion. By systematically varying the detergent concentration and characterizing the SNARE-liposome size distributions by light scattering, we describe a detailed procedure to reconstitute SNAREs into large liposomes with considerably reduced curvature stress. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 5 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Professor | 1 | 20% |
Student > Ph. D. Student | 1 | 20% |
Researcher | 1 | 20% |
Unknown | 2 | 40% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 1 | 20% |
Unknown | 4 | 80% |