Preparation, Reconstitution, and Analysis of Empty Nanodiscs
5:11
Preparation and Analysis of the Nanodisc-Monotopic Protein Complex
7:56
Results: Monotopic Membrane Protein Binding on the Surfaces of Empty Nanodiscs (NDs)
9:26
Conclusion
Transkript
The overall goal of this procedure is to visualize the binding of an extrinsic membrane protein on a nanodisc membrane surface by negative stain transmission electron microscopy as a first step towards high resolution structure determination of th
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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.