Martin-Luther-Universität Halle-Wittenberg FOR 1145 NMR studies on polyphile-lipid interactions – an update Ruth Bärenwald, Anja Achilles, Frank Lange,

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 PhysikNMR group


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


FOR 1145

B12 and clones: GUVs and MLV phase behavior

20 mmDPPC+ B12

DPPC+ E12/7 H / kcal mol-1

10.98

6.45

4.37

1.35

0.94

30 40 50 60 70 800

2

4

6

4:1

10:1

c p/

kca

l mo

l-1K

-1

temperature / °C

pure DPPC5

1:1

20:1I II III

structural model?

nature of phase transitions?

S. Werner, H. Ebert, B.-D. Lechner, F. Lange, A. Achilles, R. Bärenwald, S. Poppe, A. Blume, K. Saalwächter, 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 anX‑Shaped Bolapolyphile within Lipid Bilayers. Langmuir 31, 2839 (2015)


FOR 1145

60 30 0 -30 -60 60 30 0 -30 -60

1:4 mix

65°C

d / ppm

25°C

DPPC

d / ppm

CS

an

iso

tro

py s

/ p

pm

20 30 40 50 60 70 80

1

10

T2

/ m

stemperature / °C

1:4 mixDPPC

b)

a)

c)40

50

60

70

80

honeycomb cells filled with constrained DPPC and lateral chains

honeycomb walls, each edge formed by p-stacked BPs

DPPC-B12: head group motion and structural model

31P NMR


FOR 1145

DPPC-B12: phase transformation I

13C MAS NMR DPPC-B12 4:1

direct vs cross polarization

DP spectrum 70°C(mobile residues)

(Teflon)

Þ change of p-p packing motif at I


FOR 1145

1H MAS NMR (5 kHz spinning)DPPC-B12 4:1

DPPC-B12: phase transformations II and III

Þ alkyl chain “melting” at II

Þ B12 dissolution at III


FOR 1145

B12 core mobility and p-p packing

13C-1H dipolar MAS NMR (“DIPSHIFT”): DCH Þ SCH = <P2(cos q)>

molecular motion

CH

H H

CH

ca. 120°

ca. 180°

Þ LT: fast 180° (p) flips of phenyl rings – same as in bulk crystal Þ HT: uniaxial rotation

DPPC-B12 4:1

ar3 resonance

ar3 resonance


FOR 1145

Lipid mobility in HT phase

13C-1H dipolar MAS NMR (“DIPSHIFT”)

Þ no influence of dissolved B12 on whole-lipid motion


FOR 1145

B12 order parameters and orientation fluctuations

1H-1H dipolar MAS NMR (“BaBa-xy16”)

DPPC-B12(C16)

DOPC-B12(C18)

CH

H H

CH

ca. 120°

ca. 180°

a

DHH Þ SHH

wobbling on/in a cone

4:1 a = 20-30°10:1 a = 35-45°

10:1 a = 30-35°

Þ slight tilt of B12


FOR 1145

Why are B12-GUVs spherical?

Þ shifted fluorescence indicates -p p stacked B12 all over the

GUV

Þ filaments?


FOR 1145

E12/7: “dissolved” vs filaments

1H MAS NMR (5 kHz spinning)DPPC-E12/7 4:1

Þ E12/7 core remains immobile above main lipid phase transition


FOR 1145

E12/7: “dissolved” vs filaments

13C-1H dipolar MAS NMR (“DIPSHIFT”) DPPC-E12/7 4:1

Þ E12/7 cores are well p-p packed, some smaller-amplitude fast dynamics

Þ likely filaments!


FOR 1145

-(PGMA)m-(PPO)n-(PGMA)m- m = 20 Rmax,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)


FOR 1145

20 21 22 23 24 25 26 27 28

0

2

4

6

8

10

12

14

16

18

c

p / k

cal m

ol-1

K-1

temperature / °C

DMPC + GP34 in H2O

X:1

10

20

50

100

200

DMPC pure

20 21 22 23 24 25 26 27 28

0

2

4

6

8

10

12

14

16

18

cp /

kca

l mo

l-1 K

-1

temperature / °C

X:1

DMPC + GP12 in D2O

4

6

10

20

50

DMPC pure

20 21 22 23 24 25 26 27 28

0

2

4

6

8

10

12

14

16

18

cp /

kca

l mo

l-1 K

-1

temperature / °C

X:1

4

6

10

20

50

DMPC pure

DMPC + GP17 in D2Ofrom Schwieger et al.

Soft Matter 10, 6147 (2014)

DSC results


FOR 1145

6 5 4 3 2 1 0

ppm

DMPC/GP17 20:1

DMPC/GP34 20:1

g2

g3

g12

3

14

21

3, 3+db, ca, a’,HOD

f

e

4-13

only one useable separate polymer

resonance

1H MAS NMR


FOR 1145

mixtures with GP17:

DLPC/GP17 20:1DMPC/GP17 20:1DMPC/GP17 10:1

no cross peaks visible!

mixtures with GP34:

DMPC/GP34 20:1DMPC/GP34 60:1DMPC/GP34 150:1DPPC/GP34 50:1

cross peaks visibleÞ (some) polymers are

inserted into the bilayer

2.0 1.8 1.6 1.4 1.2 1.0 0.82.0

1.8

1.6

1.4

1.2

1.0

0.8

2

/ ppm

1 / ppm

DPPC/GP34 50:1

2.0 1.8 1.6 1.4 1.2 1.0 0.82.0

1.8

1.6

1.4

1.2

1.0

0.8

2 /

ppm

1 / ppm

DPPC/GP17 20:1

1H MAS 2D NOESY NMR


FOR 1145

BCP

lipid Re2>0

Amphi-/polyphilic tri-/pentablock copolymers

finite segmental order parameter in membrane-spanning configuration

measurable in terms of 1H-1H residual dipole-dipole coupling


FOR 1145

DQ experiments• H-H residual dipolar coupling can be determined from double-quantum build-up curve

0 2 4 6 8 10 12 14 16 18 200.0

0.2

0.4

0.6

0.8

1.0

DLPC/GP17 (50:1) DPPC/GP34 (50:1) DMPC/GP34 (60:1)

norm

aliz

ed D

Q in

tens

ity

DQ

/ ms

Mixtures with GP17:DLPC/GP17 20:1DMPC/GP17 20:1DMPC/GP17 10:1

no build-up of PPO peak => isotropic mobility

Mixtures with GP34:

Dres/2p

/ Hz

s/2p/ Hz

faniso

/ %

DMPC/GP34 20:1

130±20

45±20 18±5

DMPC/GP34 60:1

150±30

40±15 43±5

DMPC/GP34 150:1

155±40

45±30 39±5

DPPC/GP34 50:1

140±45

38±5 29±5


FOR 1145

Summary

¨ bla1¨ bla 2

also¨ bla 3

Þ also auch

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many, Many MANY FOR members...

Documents

Martin-Luther-Universität Halle-Wittenberg FOR 1145 NMR studies on polyphile-lipid interactions – an update Ruth Bärenwald, Anja Achilles, Frank Lange,