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Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy
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Biochemistry
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JoVE Journal Biochemistry
Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy

Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy

DOI:
10.3791/55148-v

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10:49 min

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March 05, 2017

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, , ,
1Department of Biosciences and Nutrition,Karolinska Institutet, 2School of Technology and Health,KTH Royal Institute of Technology

Chapters

  • 00:05Title
  • 01:08Preparation, Reconstitution, and Analysis of Empty Nanodiscs
  • 05:11Preparation and Analysis of the Nanodisc-Monotopic Protein Complex
  • 07:56Results: Monotopic Membrane Protein Binding on the Surfaces of Empty Nanodiscs (NDs)
  • 09:26Conclusion

Summary

Automatic Translation

Automatic Translation

Many proteins perform their function when attached to membrane surfaces. The binding of extrinsic proteins on nanodisc membranes can be indirectly imaged by transmission electron microscopy. We show that the characteristic stacking (rouleau) of nanodiscs induced by the negative stain sodium phosphotungstate is prevented by the binding of extrinsic protein.

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Keywords: Membrane ProteinsTransmission Electron MicroscopyNanodiscMSP1E3D1POPCSodium CholateProtein-membrane InteractionDrug DevelopmentStructural Analysis

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