1. Martin-Luther-Universität Halle-Wittenberg FOR 1145 NMR studies on polyphile-lipid interactions – an update Ruth Bärenwald, Anja Achilles, Frank Lange, Kay Saalwächter Martin-Luther-Universität Halle-Wittenberg Institut für Physik NMR group

2. Martin-Luther-Universität Halle-Wittenberg FOR 1145  Rigid-rod bola-polyphiles – phase transitions –  structural model –  evidence from NMR (and complementary methods)  Tri- and Penta-block copolymers – phase transitions – structural model and NMR evidence  Outlook NMR studies on polyphile-lipid interactions – an update Ruth Bärenwald, Anja Achilles, Frank Lange, Kay Saalwächter

3. Martin-Luther-Universität Halle-Wittenberg FOR 1145 B12 and clones: GUVs and MLV phase behavior 20  m DPPC + B12 DPPC + E12/7 IIIIII structural model? nature of phase transitions? S. Werner, H. Ebert, B.-D. Lechner, F. Lange, A. Achilles, R. Bärenwald, S. Poppe, A. Blume, K. Saalwa ̈ chter, C. Tschierske, K. Bacia, Dendritic domains with hexagonal symmetry formed by X- shaped bolapolyphiles in lipid membranes. Chem. Eur. J. 2015, DOI: 10.1002/chem.201405994. B.-D. Lechner, H. Ebert, M.Prehm, S. Werner, A. Meister, G. Hause, A. Beerlink, K. Saalwächter, K. Bacia, C. Tschierske, A. Blume, Temperature-Dependent In-Plane Structure Formation of an X ‑ Shaped Bolapolyphile within Lipid Bilayers. Langmuir 31, 2839 (2015)

4. Martin-Luther-Universität Halle-Wittenberg FOR 1145 honeycomb cells filled with constrained DPPC and lateral chains honeycomb walls, each edge formed by  -stacked BPs DPPC-B12: head group motion and structural model 31 P NMR

5. Martin-Luther-Universität Halle-Wittenberg FOR 1145 DPPC-B12: phase transformation I 13 C MAS NMR DPPC-B12 4:1 direct vs cross polarization DP spectrum 70°C (mobile residues) (Teflon)  change of  -  packing motif at I

6. Martin-Luther-Universität Halle-Wittenberg FOR 1145 1 H MAS NMR (5 kHz spinning) DPPC-B12 4:1 DPPC-B12: phase transformations II and III  alkyl chain “melting” at II  B12 dissolution at III

7. Martin-Luther-Universität Halle-Wittenberg FOR 1145 B12 core mobility and  -  packing 13 C- 1 H dipolar MAS NMR (“DIPSHIFT”): D CH  S CH =  P 2 (cos  )  molecular motion C H H H C H ca. 120° ca. 180°  LT: fast 180° (  ) flips of phenyl rings – same as in bulk crystal  HT: uniaxial rotation DPPC-B12 4:1 ar 3 resonance

8. Martin-Luther-Universität Halle-Wittenberg FOR 1145 Lipid mobility in HT phase 13 C- 1 H dipolar MAS NMR (“DIPSHIFT”)  no influence of dissolved B12 on whole-lipid motion

9. Martin-Luther-Universität Halle-Wittenberg FOR 1145 B12 order parameters and orientation fluctuations 1 H- 1 H dipolar MAS NMR (“BaBa-xy16”) DPPC-B12 (C 16 ) DOPC-B12 (C 18 ) C H H H C H ca. 120° ca. 180°  D HH  S HH wobbling on/in a cone 4:1  = 20-30° 10:1  = 35-45° 10:1  = 30-35°  slight tilt of B12

10. Martin-Luther-Universität Halle-Wittenberg FOR 1145 Why are B12-GUVs spherical?  shifted fluorescence indicates  stacked B12 all over the GUV  filaments?

11. Martin-Luther-Universität Halle-Wittenberg FOR 1145 E12/7: “dissolved” vs filaments 1 H MAS NMR (5 kHz spinning) DPPC-E12/7 4:1  E12/7 core remains immobile above main lipid phase transition

12. Martin-Luther-Universität Halle-Wittenberg FOR 1145 E12/7: “dissolved” vs filaments 13 C- 1 H dipolar MAS NMR (“DIPSHIFT”) DPPC-E12/7 4:1  E12/7 cores are well  -  packed, some smaller-amplitude fast dynamics  likely filaments!

13. Martin-Luther-Universität Halle-Wittenberg FOR 1145 -(PGMA) m -(PPO) n -(PGMA) m - m = 20 R max,PPO GP12: n = 12 3.5 nm (new) GP17: n = 17 5 nm (new) GP34: n = 34 10 nm mixed with: DLPC DMPC DPPC 12 14 16 Amphi-/polyphilic tri-/pentablock copolymers C. Schwieger, A. Achilles, S. Scholz, J. Rüger, K. Bacia, K. Saalwächter, J. Kressler, A. Blume. Binding of amphiphilic and triphilic block copolymers to lipid model membranes: the role of perfluorinated moieties. Soft Matter 10, 6147 (2014)

14. Martin-Luther-Universität Halle-Wittenberg FOR 1145 from Schwieger et al. Soft Matter 10, 6147 (2014) DSC results

15. Martin-Luther-Universität Halle-Wittenberg FOR 1145 g2g2 g3g3 g1g1 2 3 14 2 1 3, 3+d b, c a, a’, HOD f e 4-13 only one useable separate polymer resonance 1 H MAS NMR

16. Martin-Luther-Universität Halle-Wittenberg FOR 1145 mixtures with GP17: DLPC/GP17 20:1 DMPC/GP17 20:1 DMPC/GP17 10:1 no cross peaks visible! mixtures with GP34: DMPC/GP34 20:1 DMPC/GP34 60:1 DMPC/GP34 150:1 DPPC/GP34 50:1 cross peaks visible  (some) polymers are inserted into the bilayer 1 H MAS 2D NOESY NMR

17. Martin-Luther-Universität Halle-Wittenberg FOR 1145 BCP lipid R e 2 >0 Amphi-/polyphilic tri-/pentablock copolymers finite segmental order parameter in membrane-spanning configuration measurable in terms of 1 H- 1 H residual dipole-dipole coupling

18. Martin-Luther-Universität Halle-Wittenberg FOR 1145 DQ experiments H-H residual dipolar coupling can be determined from double-quantum build-up curve Mixtures with GP17: DLPC/GP17 20:1 DMPC/GP17 20:1 DMPC/GP17 10:1 no build-up of PPO peak => isotropic mobility Mixtures with GP34: D res /2  / Hz  /2  / Hz f aniso / % DMPC/GP34 20:1 130  2045  2018  5 DMPC/GP34 60:1 150  3040  1543  5 DMPC/GP34 150:1 155  4045  3039  5 DPPC/GP34 50:1 140  4538  529  5

19. Martin-Luther-Universität Halle-Wittenberg FOR 1145 Summary  bla1  bla 2  also  bla 3  also auch ! €€€: thanks to: many, Many MANY FOR members...

20. Martin-Luther-Universität Halle-Wittenberg FOR 1145 O O O O OH HO bolapoly- phile DPPC lipid