Fig. 1. The sequence and α-helical structure of melittin

Fig. 1. Diffusion of lipids on a DOPC/DPPC membrane under the action of melittin at 0.5 µg/mL: (a) Microscopy image with some trajectories marked in colors; (b) typical lipid trajectories in 3 s; Some of the immobile ones are marked with blue arrows; (c) representative time-averaged MSD of lipids.

Fig. 2. A2. Molecular structure of DOPC, DPPC and Rh-PE

Fig. 2. PDF distribution of lipid diffusion coefficients (D_L) of membrane with different melittin concentrations: (a) Histograms andfittings of the PDF; (b) corresponding peak locations and area proportionsin (a). Sample numbers are 657 (pristine membrane), 427 (with Mel at 0.5 µg/mL) and 507 (with Mel at 5.0 µg/mL), respectively.

Fig. 3. Three different types of lipid diffusion modes and their PDFs in three conditions of membrane without or with melittin exposure. Representative trajectories are shown on the right.

Fig. 3. Confocal images of calcein-encapsulated GUVs exposed to melittin at 0.5 or 5.0 µg/mL. The images were taken in the green (calcein), red (lipid) and overlaid channels. Cartoons on the right refer to the corresponding action states of peptides, including surface adsorption and transmembrane insertion

Fig. 4. PDF of lipids in a pure DOPC bilayer. The data were obtained from three times of independently repeated tests, with a sample number of 404. Peak of the fitted curve locates at 2.780 µm²/s

Fig. 4. Interaction between melittin and a phase-separated bilayer: (a) Snapshots showing the melittin-inducedporeformation process. Top: top view, bottom: side view. Red: melittin, blue: DPPC headgroup, green: DUPC headgroup. For clarity, lipid tails are not shown; (b) interaction details between Mel (red) and the surrounding lipids. Color codes are the same as in (a), with tails of the surrounding lipids in yellow, and heads of lipids away from the peptides in grey; (c) time evolution of phase boundary length (L) before and after the addition of melittin at P/L = 12/512. The time points of (a) are marked correspondingly in (c).

Fig. 5. Example of lipid trajectories handled by wavelet:　(a) X-coordinate profile of the original trajectory;(b) wavelet coefficients from the 2 nd to the 64 th

Fig. 6. The wavelet coefficients of the 2 nd, 32 th and 64 th scale; (d) the 32 th wavelet coefficient and the corresponding red line of threshold value

Fig. 7. (a) Trajectory discriminated by wavelet using the 32 th scale coefficient, whose red part refers to the “fast” motion type and blue part refers to the “slow” motion type;(b)the corresponding MSD distributions of the red and blue parts.

Fig. 8. Changes in MSD distributions of lipids before and after the addition of Mel in the simulations