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. 2020 Aug 7;11:3944. doi: 10.1038/s41467-020-17585-8

Fig. 2. Partitioning of hydrophobic and amphipathic helix-containing proteins to droplet interface bilayers.

Fig. 2

a Formation of protein-containing DIBs: soluble protein, proteoliposome, or purified LDs are added to the buffer droplet (left). Mixing the droplet in a TG-containing phospholipids solution allows the relocalization of proteins to the buffer droplet interface (middle). When two drops come close together, their monolayers zip to form a bilayer. The proteins are thus offered to relocalize to the bilayer (right). b Protein distribution between the bilayer and the monolayer is determined by the partition coefficient P, which is the protein signal at the bilayer divided by sum of its signals at the monolayers. When a protein partitions preferentially to the bilayer, p > 1; if preference is for the monolayer then p < 1. c, d Distribution of GMAP-210-AH (soluble protein) and ACSL3 (monotopic membrane protein) respectively, in DOPE or DOPC/DOPE (1:1) DIBs. Scale bar: 20 µm. The partition coefficient is represented for each condition as box-plots from n = 5 independent measurements (excepted for GMAP-210-AH in PE, n = 4). e Enrichment parameter in DOPC/DOPE (1:1) membranes for AH- (green) and HD- (pink) containing proteins, shown as floating bars (bar limits, min to max values; central line, mean), 2 ≤ n ≤ 8 independent measurements were done for each protein. HD-containing proteins coming from LDs are Plin 1, Plin 1C, ACSL3, CG2254, CG9186, Oleosin 1, Caveolin 1, HPos; those coming from proteoliposomes are Syt1 57-421, t-snare, Vamp2; AH-containing proteins coming from LDs are Plin3, Plin2, Plin1N; the other AHs are added soluble. f Average partition coefficient of the groups of soluble or monotopic membrane proteins in DOPE or DOPC/DOPE (1:1). Results presented in (Supplementary Fig. S2e) were used to generate (f). Box-plots are defined as follow: box limits, upper and lower quartiles; middle line, median; whiskers, minimum and maximum value. In c, d, e, f each point is represented as a black dot. Source data are provided as a Source Data file.