Figure 3 | Scientific Reports

Figure 3

From: Structure based biophysical characterization of the PROPPIN Atg18 shows Atg18 oligomerization upon membrane binding

Figure 3

(A) Atg18 structure showing residues, which were mutated to cysteines for fluorescence labeling experiments. E170 is marked with an asterisk because its side chain is disordered in the structure and was modelled as an alanine. (B) Liposome flotation assays were performed with wild-type, cysteine-free and single cysteine Atg18 mutants using 100 nm LUVs composed of DOPC/DOPE/Texas Red-PE/PtdIns(3,5)P2 (76:21:2:1, molar ratio). The top two fractions of the gradient show protein bound to liposomes and bottom fractions contain unbound protein. The S448C FTTG mutant served as a negative control. (C) Observed changes of the normalized fluorescence at 520 nm for the Oregon Green labelled mutants after Texas Red liposome addition. (D) Model for Atg18 binding at the membrane. The docking was manually performed taking data from (C) into account. Residues are color coded according to the observed change in normalized fluorescence. Color bar is shown.