²H NMR spectra and order parameters of DMPC-<i>d</i>₅₄/DMPS membranes in the absence and presence of ghrelin and deacylghrelin.

<p>²H NMR spectra in DMPC-<i>d</i>₅₄/DMPS membranes (5/1, mol/mol) in the presence of ghrelin (A) and ghrelin-<i>d</i>₁₅ in DMPC/DMPS membranes (B). C) ²H NMR order parameters of DMPC-<i>d</i>₅₄/DMPS (5:1, mol/mol) membranes in the presence or absence of ghrelin or desacylghreliln (1:30 protein to lipid molar ratio) at a temperature of 30°C and a buffer content of 35 wt%. Error bars of the ²H NMR order parameters are smaller than the symbol size.</p

Chemical shifts measured for acylated ghrelin bound to DMPC/DMPC membranes (5/1, mol/mol) using MAS solid-state NMR.

<p>Chemical shifts measured for acylated ghrelin bound to DMPC/DMPC membranes (5/1, mol/mol) using MAS solid-state NMR.</p

Ghrelin sequence showing the isotopically labeled amino acids of the different ghrelin molecules and ssNMR spectra of membrane-embedded ghrelin.

<p>Labeled amino acids are shown in bold italics (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122444#pone.0122444.s010" target="_blank">S2 Table</a>). A) ¹³C CP MAS NMR spectrum of ghrelin (with Gly1, Leu5, and Ser6 ¹³C/¹⁵N labeled) in DMPC-<i>d</i>₆₇/DMPS-<i>d</i>₅₄ (5:1, mol/mol) membranes at a ghrelin concentration of 3.3 mol%. B) ¹H-¹³C MAS HetCor NMR spectrum of the same preparation, all at 30°C and a MAS frequency of 7 kHz.</p

Outline of model ensemble selection algorithm.

<p>The flowchart outlines the process by which the agreement with experimental data is determined for an ensemble of models selected from a large pool.</p

Secondary structure analysis of ghrelin.

<p>A) Ramachandran plots of various subsets of residues as labeled at the top of the plots. The torsion angles of all models generated in Rosetta (gray) and the final ensemble of models (black) are plotted. B) Weblogo (<a href="http://weblogo.threeplusone.com" target="_blank">http://weblogo.threeplusone.com</a>) of PPII-DSSP analysis of final ensemble of ghrelin models. Color key: black = random coil (C), blue = bend (S) or turn (T), and green = α-, 3₁₀-, or PPII helix (H, G, or P, respectively).</p