Figure 3.

Cartoon schematic of proteoliposome sample preparation for MAS-DNP. The lipid/peptide multilamellar lipid vesicle (MLVs) are produced following traditional membrane peptide or protein sample preparations for ssNMR. Here, the lipid pellet is first hydrated in excess buffer and subjected to 15X freeze/thaw cycles to form homogenous MLVs and pelleted by ultracentrifugation. Following this, steps 1–3 illustrate addition of polarizing agents (PA), and additional freeze/thaw cycles to distribute the PA and glassing agent prior to sample transfer into the NMR rotor.