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| Title | Lipid nanodiscs as a template for high-resolution cryo-EM structures of peripheral membrane proteins. |
|---|---|
| Journal, issue, pages | J Struct Biol, Vol. 215, Issue 3, Page 107989, Year 2023 |
| Publish date | Jun 24, 2023 |
 Authors | Kevin S Cannon / Reta D Sarsam / Tanita Tedamrongwanish / Kevin Zhang / Richard W Baker /  |
| PubMed Abstract | Peripheral membrane proteins are ubiquitous throughout cell biology and are required for a variety of cellular processes such as signal transduction, membrane trafficking, and autophagy. Transient ...Peripheral membrane proteins are ubiquitous throughout cell biology and are required for a variety of cellular processes such as signal transduction, membrane trafficking, and autophagy. Transient binding to the membrane has a profound impact on protein function, serving to induce conformational changes and alter biochemical and biophysical parameters by increasing the local concentration of factors and restricting diffusion to two dimensions. Despite the centrality of the membrane in serving as a template for cell biology, there are few reported high-resolution structures of peripheral membrane proteins bound to the membrane. We analyzed the utility of lipid nanodiscs to serve as a template for cryo-EM analysis of peripheral membrane proteins. We tested a variety of nanodiscs and we report a 3.3 Å structure of the AP2 clathrin adaptor complex bound to a 17-nm nanodisc, with sufficient resolution to visualize a bound lipid head group. Our data demonstrate that lipid nanodiscs are amenable to high-resolution structure determination of peripheral membrane proteins and provide a framework for extending this analysis to other systems. |
 External links | J Struct Biol / PubMed:37364761 |
| Methods | EM (single particle) |
| Resolution | 3.3 - 6.8 Å |
| Structure data |  EMDB-16803: AP2 bound to MSP1 nanodisc with Tgn38 cargo peptide  EMDB-16804: AP2 bound to MSP1E3D1 nanodisc with Tgn38 cargo peptide  EMDB-29977: AP2 bound to an MSP1 nanodisc  EMDB-40034: Structure of AP2 bound to MSP1E3D1 nanodisc  EMDB-40035: Structure of AP2 bound to MSP2N2 nanodisc  EMDB-40963: AP2 bound to MSP2N2 nanodisc with Tgn38 cargo peptide; focused refinement 1  EMDB-40964: AP2 bound to MSP2N2 nanodisc with Tgn38 cargo peptide; focused refinement 2  EMDB-40965: AP2 bound to MSP2N2 nanodisc with Tgn38 cargo peptide; focused refinement 3  EMDB-40966: AP2 bound to MSP2N2 nanodisc with Tgn38 cargo peptide; consensus refinement EMDB-40973, PDB-8t1o: |
| Chemicals |  ChemComp-PIO: |
| Source |
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 Keywords |  ENDOCYTOSIS / Clathrin-mediated endocytosis; peripheral membrane protein |
