Prospects & Overviews
Studying protein-reconstituted proteoliposome fusion with content indicators in vitro
Article first published online: 29 APR 2013
© 2013 WILEY Periodicals, Inc.
Volume 35, Issue 7, pages 658–665, July 2013
How to Cite
Diao, J., Zhao, M., Zhang, Y., Kyoung, M. and Brunger, A. T. (2013), Studying protein-reconstituted proteoliposome fusion with content indicators in vitro. Bioessays, 35: 658–665. doi: 10.1002/bies.201300010
- Issue published online: 13 JUN 2013
- Article first published online: 29 APR 2013
- Central Universities (Xi'an Jiaotong University to J.D.) and the National Institutes of Health (R37-MH63105 to A.T.B.)
- content mixing;
- membrane fusion;
- single molecule;
In vitro reconstitution assays are commonly used to study biological membrane fusion. However, to date, most ensemble and single-vesicle experiments involving SNARE proteins have been performed only with lipid-mixing, but not content-mixing indicators. Through simultaneous detection of lipid and small content-mixing indicators, we found that lipid mixing often occurs seconds prior to content mixing, or without any content mixing at all, during a 50-seconds observation period, for Ca2+-triggered fusion with SNAREs, full-length synaptotagmin-1, and complexin. Our results illustrate the caveats of commonly used bulk lipid-mixing fusion experiments. We recommend that proteoliposome fusion experiments should always employ content-mixing indicators in addition to, or in place of, lipid-mixing indicators.