Extended Data Fig. 5: The role of charge and hydrophobicity in phospholipid partitioning.
From: Biomolecular condensates create phospholipid-enriched microenvironments
a, Chemical library used for studying the effect of length and headgroup on phospholipid enrichment in MED1 condensates. b, Condensate metabolome experiments were performed using the chemical library from d rather than liver metabolite extract, with each molecule added to a final concentration of 100 nM (blue), 1 μM (green) or 10 μM (orange). The role of phospholipid tail length was assessed by plotting the combined tail length (x-axis) of each phosphatidylcholine against their log2-fold enrichment in MED1 condensates (y-axis). Choline glycerophosphoryl head group (carbons = 0) and the library’s lysophosphatidylcholine (carbons = 16) were also included. Results from each replicate are plotted as separate dots and crossbars represents means across replicates. Enrichment is only apparent with fatty acyl moiety chain length > 15. Combined fatty acyl moieties lengths of 16 and 18 have reduced enrichment at the lowest concentration, while phospholipids with longer fatty acyl moieties (>23) have higher enrichment at lower concentrations. Error bars indicate s.e.m. (n = 3). c, Condensate enrichment was compared between phospholipids with dipalmitoyl fatty acyl moieties, but different head groups using the library described in a. The log2-fold enrichment in MED1 condensates (y-axis) is plotted for each phospholipid (x-axis) when metabolites were added 100 nM (blue), 1 μM (green) or 10 μM (orange). Results of individual replicates are plotted as dots (black), while bars indicate the mean. Error bars indicate s.e.m. (n = 3). d, Net neutral charge phospholipids preferentially partition into MED1 and HNRNPA1 condensates. Phospholipids from the metabolomics datasets analyzed in Fig. 2 were grouped based on head group (x-axis): sphingomyelins (SM, n = 4), phosphatidylcholines (PC, n = 4), phosphatidylethanolamines (PE, n = 27), phosphatidylinositols (PI, n = 5), and phosphatidylserines (PS, n = 11). For each group, median log2-fold enrichment was plotted for nucleocapsid (purple), MED1 (blue; p = 0.02637, SM/PS; p = 0.02584, PE/PI; p = 0.0001876, PE/PS) or HNRNPA1 (green; p = 0.01587, SM/PI; p = 0.001465, SM/PS, p = 0.01587, PC/PI; p = 0.001465, PC/PS; p = 0.00149, PE/PI; p = 6.79e-06, PE/PS) condensates. Violin plots represent the distribution of median enrichment for each head group and lines demarcate quartiles. *p < 0.05, **p < 0.005, two-sided Wilcoxon rank-sum test (n = 3 condensate metabolomic experiments per protein).
