| Analyzing membrane remodeling and fission using supported bilayers with excess membrane reservoir. | |
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MedLine Citation:
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PMID: 23288321 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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A complete understanding of the molecular mechanisms governing vesicle formation requires quantitative assays and vesicle reconstitution using purified components. We describe a simple model membrane template for studying protein-mediated membrane remodeling and vesicle formation or fission that is amenable to both quantitative biochemical analysis and real-time imaging by epifluorescence microscopy. Supported bilayers with excess membrane reservoir (SUPER) templates are compositionally well-defined unilamellar membrane systems prepared on 2-5-μm silica beads under conditions that enable incorporation of excess membrane to form a loosely fitting bilayer that can be used to study membrane remodeling and fission. This protocol describes methods for SUPER template formation and characterization, as well as for the qualitative observation and quantitative measurement of vesicle formation and fission via microscopy and a simple sedimentation assay. SUPER templates can be prepared within 60 min. Results from either sedimentation-based or microscopy-based assays can be obtained within an additional 60 min. |
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Authors:
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Sylvia Neumann; Thomas J Pucadyil; Sandra L Schmid |
Publication Detail:
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Type: Journal Article Date: 2013-01-03 |
Journal Detail:
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Title: Nature protocols Volume: 8 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2013 Jan |
Date Detail:
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Created Date: 2013-01-04 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
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Languages: eng Pagination: 213-22 Citation Subset: IM |
Affiliation:
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Department of Cell Biology, The Scripps Research Institute, La Jolla, California, USA. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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