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Co-Translational Insertion of Membrane Proteins into Preformed Nanodiscs
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Biochemistry
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JoVE Journal Biochemistry
Co-Translational Insertion of Membrane Proteins into Preformed Nanodiscs

Co-Translational Insertion of Membrane Proteins into Preformed Nanodiscs

DOI:
10.3791/61844-v

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08:24 min

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November 19, 2020

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, , ,
1Institute of Biophysical Chemistry,Goethe University Frankfurt

Chapters

  • 00:05Introduction
  • 00:45S30 Ribosomal Protein Lysate Preparation
  • 02:17Membrane Scaffold Protein 1E3D1 (MSP1E3D1) Nanodisc Assembly
  • 03:243-mL Continuous Exchange Cell-Free (CECF) Reaction Setup
  • 04:1060-Microliter CECF Reaction Setup
  • 06:07Results: Representative Preformed Nanodisc-Modified Membrane Protein Characterization
  • 07:40Conclusion

Summary

Automatic Translation

Automatic Translation

Co-translational insertion into pre-formed nanodiscs makes it possible to study cell-free synthesized membrane proteins in defined lipid environments without contact with detergents. This protocol describes the preparation of essential system components and the critical parameters for improving expression efficiency and sample quality.

Tags

Keywords: Co-translational InsertionMembrane ProteinsNanodiscDetergent-freeProtein FoldingLipid EffectsCell-free ExpressionMembrane Scaffold ProteinCell DisruptionDialysis

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