Supported bilayers with excess membrane reservoir: a template for reconstituting membrane budding and fission.

Biophysical Journal
Thomas J Pucadyil, Sandra L Schmid

Abstract

A complete mechanistic understanding of membrane-localized processes in vesicular transport, such as membrane budding and fission, requires their reconstitution with biochemically-defined components from a biochemically-defined substrate. Supported bilayers formed by vesicle fusion represent an attractive substrate for this purpose. However, conventional supported bilayers lack a sufficient membrane reservoir to recreate membrane budding and fission events. We describe the formation of supported bilayers with excess membrane reservoir (SUPER) templates from the fusion of liposomes containing negatively charged lipids on silica beads under high-ionic-strength conditions. Using a fluorescence microscopy-based assay to monitor early and late stages of supported bilayer formation, we show that an increase in ionic strength leads to an increase in the rates of liposome adsorption and subsequent fusion during formation of supported bilayers. The two rates, however, increase disproportionally, leading to accumulation of excess reservoir with an increase in ionic strength. SUPER templates allow the seamless application of microscopy-based assays to analyze membrane-localized processes together with sedimentation-based assays to isolate...Continue Reading

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Citations

Jun 15, 2011·Proceedings of the National Academy of Sciences of the United States of America·Ya-Wen LiuSandra L Schmid
Mar 23, 2013·Science·Anna V ShnyrovaVadim A Frolov
Feb 6, 2013·PloS One·Ya-Wen LiuSandra L Schmid
May 24, 2011·Annual Review of Cell and Developmental Biology·Sandra L Schmid, Vadim A Frolov
Nov 26, 2015·The Journal of Cell Biology·Caitlin Sedwick, Caitlin Sedwick
May 10, 2018·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Michael G HannaAnjon Audhya
Nov 17, 2017·Scientific Reports·Javier H HervásAlicia Alonso
Nov 1, 2018·Experimental Biology and Medicine·Sarina ChandBarbara Ciani
Apr 4, 2015·The Journal of Biological Chemistry·Sachin S HolkarThomas J Pucadyil
Aug 11, 2016·Soft Matter·Jaime Agudo-Canalejo, Reinhard Lipowsky
Dec 31, 2016·Molecular Biology of the Cell·Srishti Dar, Thomas J Pucadyil
May 6, 2017·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Morgan ChabanonPadmini Rangamani
Jul 19, 2013·The Journal of Biological Chemistry·Sylvia Neumann, Sandra L Schmid
Sep 29, 2018·Journal of Cell Science·Sagardip MajumderG W Gant Luxton
Jul 18, 2018·Chemistry and Physics of Lipids·Noah A SchenkArun Anantharam
Jul 11, 2021·Proceedings of the National Academy of Sciences of the United States of America·Oleg M GanichkinJeffrey K Noel
Nov 8, 2018·Biochemistry·Sukrut C KamerkarThomas J Pucadyil

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