Fig 3. MD simulations to investigate EDTA aggregation and interaction with SYPRO Orange.

(A) SASA and frequency distribution of the SASA over all 20 EDTA molecules in the simulation box; EDTA^4- (black), EDTA^3- (red), the arrow shows which snapshot was used to visualize the EDTA aggregate. (B) Snapshot of an MD trajectory showing an EDTA^4—Na⁺ aggregate; the snapshot was taken at 500 ns (see arrow in panel A); EDTA^4-: green: C atoms, blue: N atoms, red: O atoms; magenta: Na⁺ ions. Box: This part of the image was zoomed to get a better view on the bilayer within the aggregate. For clarity, some EDTA molecules in front of the bilayer were discarded. (C) Zoomed view on an EDTA^4—Na⁺ aggregate showing a bilayer formation.