File S1 - Structure Analysis and Conformational Transitions of the Cell Penetrating Peptide Transportan 10 in the Membrane-Bound State

Figure S1, CD spectra of the CF3-Bpg labeled TP10 analogs bound to DMPC/DMPG vesicles. (A) L- and (B) D-epimers at P/L = 1∶50. (C) L- and (D) D-epimers at P/L = 1∶100. The wild type TP10 is shown as a black line; green lines show analogs with CF3-Bpg labels in the galanin part, and red lines in the mastoparan part. Figure S2, CD spectra of the carboxyfluorescein- and CF3-Bpg labeled TP10 analogs in DMPC/DMPG vesicles at P/L = 1∶50. (A) L- and (B) D-epimers. The wild type TP10 is shown as a black line; green lines show analogs with CF3-Bpg labels in the galanin part, and red lines in the mastoparan part. Figure S3, Comparison of the solid-state 19F-NMR spectra of L-CF3-Bpg labeled TP10. Analogs with and without the carboxyfluorescein-label were measured in oriented DMPC/DMPG bilayers at P/L = :200, at 40°C, with the sample normal aligned parallel (0°) and perpendicular (90°) to the static magnetic field. Figure S4, Internalization of the carboxyfluorescein-labeled TP10-WT and the L- and D-CF3-Bpg analogs by HeLa-cells. The cells were incubated with 2 µM (left column) and 10 µM (right column) peptide at 37°C for 30 min. Figure S5, Solid-state 19F-NMR spectra of the L-CF3-Bpg labeled TP10. Analogs were measured in oriented DMPC/DMPG bilayers at P/L = 1∶50 and 1∶200, at 40°C, with the sample normal aligned parallel (0°) and perpendicular (90°) to the static magnetic field B0. Several spectra showed an immobilized powder component, as indicated by the boxes. Figure S6, OCD spectra of the D-CF3-Bpg labeled TP10. Analogs were measured in oriented DMPC/DMPG bilayers at P/L = 1∶50. Spectra were recorded after 1 (straight lines), 5 (dashed lines) and 8 days (dotted lines) hydration of the OCD sample. Figure S7, OCD spectra of the Leu16→L-CF3-Bpg labeled TP10. Peptide measured in oriented DMPC/DMPG bilayers at P/L = 1∶50. Spectra were recorded after 1 (straight lines), 5 (dashed lines) and 8 days (dotted lines) hydration of the OCD sample. Scheme S1, Chemical structure of L-CF3-Bpg (left) and D-CF3-Bpg (right). Table S1, Sequences and LC-MS characterization of TP10-WT and the analogues labeled with L-CF3-Bpg and with an additional carboxyfluorescein (CF). (Equivalent data were obtained for the D-epimers). Table S2, Comparison of the 19F-NMR dipolar couplings of the L-CF3-Bpg labeled TP10 analogs with and without the fluorescent carboxyfluorescein label in oriented DMPC/DMPG bilayers (P/L = 1∶200), measured at 0° and 90° sample tilt. Table S3, Dipolar couplings of the L-CF3-Bpg labeled TP10 analogs in oriented DMPC/DMPG bilayers, measured at 0° and 90° sample tilt, at P/L = 1∶50, 1∶200, and 1∶400. Table S4, Dipolar couplings of the D-CF3-Bpg labeled TP10 analogs in oriented DMPC/DMPG bilayers, measured at 0° and 90° sample tilt, at P/L = 1∶50, 1∶200, and 1∶400. (DOCX)