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

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

DOI:
10.3791/59867-v

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08:43 min

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July 03, 2019

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,
1Department of BioSciences,Rice University

Chapters

  • 00:04Title
  • 00:56Liposome Production by Extrusion Method
  • 02:45Reconstitution by Detergent Assisted Incorporation into Preformed Liposomes
  • 05:10Lipid Mixing Assays
  • 06:16Results: Analysis of Liposomes Reconstituted from Recombinant Drosophila Atlastin
  • 07:58Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

Keywords: Detergent-assisted ReconstitutionRecombinant Drosophila AtlastinLiposomesLipid-mixing AssaysFRET-based Lipid-mixingProteoliposomesLipid FilmFreeze-thawExtrusionLipid Concentration

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