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Biochemistry

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JoVE Journal Biochemistry
Detergent-assisted Reconstitution of Recombinant Drosophila Atlastin into Liposomes for Lipid-mixing Assays
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Detergent-assisted Reconstitution of Recombinant Drosophila Atlastin into Liposomes for Lipid-mixing Assays

Article DOI: 10.3791/59867-v • 08:43 min • July 3rd, 2019
July 3rd, 2019 •
1, 1
1Department of BioSciences, Rice University

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Biological membrane fusion is catalyzed by specialized fusion proteins. Measuring the fusogenic properties of proteins can be achieved by lipid mixing assays. We present a method for purifying recombinant Drosophila atlastin, a protein that mediates homotypic fusion of the ER, reconstituting it to preformed liposomes, and testing for fusion capacity.

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Detergent-assisted Reconstitution Recombinant Drosophila Atlastin Liposomes Lipid-mixing Assays Membrane-bound Proteins Model Lipid Bilayer Fusion Proteins FRET-based Assay Protein Reconstitution Reconstitution Orientation Lipid Mixes Chloroform DPPC Lipid Film A100 Glycerol 2-mercaptoethanol
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