Supplementary Information
Versatile peptide rafts for conjugate morphologies by self-assembling of
amphiphilic helical peptides
Motoki Ueda, Akira Makino, Tomoya Imai, Junji Sugiyama, & Shunsaku Kimura*
Index
1. Materials and Methods
2. Synthesis
3.
Molecular Assemblies Prepared from S22L16
TEM Observation (Fig. S1 and S2)
4.
Molecular Assemblies Prepared from S24L14 and S25D12
TEM Observation (Fig. S3)
ABC-Type Dumbbell Assembly
5.
6.
TEM Observation (Fig. S4)
Three-Way Nanocapsule Assembly
TEM Observation (Fig. S5)
1
1. Materials and Methods
All reagents and solvents were purchased commercially and used as-received unless
otherwise noted.
2. Synthesis
Amphiphilic polypeptides of Sar25-b-(D-Leu-Aib)6 (S25D12), Sarn-b-(L-Leu-Aib)6
(SnL12), (n = 10, 25), Sar24-b-(L-Leu-Aib)7 (S24L14), and Sar22-b-(L-Leu-Aib)8
(S22L16) were synthesized as previously reported16,19. The syntheses of all compounds
were confirmed by 1H-NMR experiments and MALDI-TOF MASS analysis.
2
3. Molecular Assemblies Prepared from S22L16
Morphology of molecular assemblies prepared from S22L16 was observed by TEM and
cryo-TEM. A S22L16 ethanol solution (0.5 mg/ 10 μL) was injected into a 10 mM TrisHCl buffer (pH 7.4) (1 mL) and the dispersion was heated at 90 °C for 1 h. TEM images
of molecular assemblies of S22L16 were obtained before and after heat treatment (Fig.
S1). S22L16 formed round planar sheets having ca. 150 nm diameter with wide
dispersity before heating. On the other hand, after heat treatment, awkward vesicles
were observed. The vesicles had ca. 70 nm diameter with a narrow size distribution.
Figure S1. TEM images (negative staining with uranyl acetate; a–c, and
cryogenic TEM; d–f) of molecular assemblies from S22L16. Before heat
treatment; a–c and after heat treatment; d–f.
3
Morphology of molecular assemblies prepared from a mixture of S22L16 sheets and
S25L12 nanotubes was observed by TEM and cryo-TEM. An aliquot (25, 50, 100 μL)
of S22L16 sheets suspension (0.5 mg/ 1 mL) was incubated with an aliquot (50 μL)
S25L12 nanotube suspension (0.5 mg/ 1 mL) and the dispersion was heated at 90 °C for
1 h. TEM images of molecular assemblies at each mixing ratio were obtained (Fig. S2).
A mixture of S22L16 sheet and S25L12 nanotube with a ratio 0.5/1 (v/v) formed roundbottom test tube as a major fraction. Data of mixing ratio of 1/1 were not shown. In the
case of mixing ratio 2/1, ABA-type nanocapsules, where the both mouths of S25L12
nanotube were sealed with round-bottom shaped S22L16 membrane, were obtained
mainly.
Figure S2. TEM images (cryogenic TEM; a–c and negative staining with uranyl
acetate; d–f) of molecular assemblies from mixtures of S22L16 sheets and
S25L12 tubes. Mixing ratio of S22L16 against S25L12, 0.5/1 ; a–c and 2/1; d–f.
4
4. Molecular Assemblies Prepared from S24L14 and S25D12
Morphology of molecular assemblies prepared from a mixture of S24L14+S25D12
nanotubes19 and S25L12+S25D12 planar sheets was observed by TEM and cryo-TEM.
First, two kind of sheets were prepared by injections of a mixture of S24L14 and
S25D12 in ethanol (0.25 mg, 0.25 mg/ 10 μL) and a mixture S25L12 and S25D12 in
ethanol (0.25 mg, 0.25 mg/ 10 μL) into 10 mM Tris-HCl buffer (pH 7.4) (1 mL),
separately. Planar sheet suspension of S24L14 and S25D12 was heated at 90 °C 1 h to
transform into large nanotubes. An aliquot (50 μL) of nanotube dispersion was
incubated with an aliquot (50 μL) of S25L12 and S25D12 nanotube dispersion and the
mixture was heated at 90 °C for 1 h. TEM images of molecular assemblies were
obtained (Fig. S3).
Figure S3. TEM images (negative staining with uranyl acetate; a–c and
cryogenic TEM; d) of molecular assemblies from a mixture of S24L14 and
S25D12 nanotubes and S25L12 and S25D12 planar sheets.
5
5. ABC-Type Asymmetric Dumbbell Assembly
Morphology of molecular assemblies prepared from a mixture of S25L12+S25D12
(molar ratio; 2/8) round-bottom flask17 and S22L16 planar sheets was observed by TEM
and cryo-TEM. First, the round-bottom flask assemblies were prepared from a mixture
of S25L12 and S25D12 at the molar ratio of 2/8 in ethanol (0.1 mg, 0.4 mg/ 10 μL) by
the ethanol injection method and heat treatment at 90 °C for 1 h. This dispersion was
purified by a syringe filter through elimination of assemblies larger than 500 nm.
Subsequently, S22L16 planar sheets dispersion (50 μL) were added to the round-bottom
flask dispersion (50 μL) and heated at 90 °C for 1 h. TEM images of molecular
assemblies were obtained (Fig. S4).
Figure S4. TEM images (negative staining with uranyl acetate; a-g and
cryogenic TEM; h) of molecular assemblies from mixtures of S25L12 and
6
S25D12 round-bottom flasks and S22L16 planar sheets with a mixing ratio of
1/1 (v/v).
7
Three-Way Nanocapsule Assembly
Morphology of molecular assemblies prepared from a mixture of S25L12+S10L12
(mixing ratio; 1/1 v/v) three-way nanotubes and S22L16 planar sheets was observed by
TEM. First, the three-way nanotube assemblies were prepared from a mixture of
S25L12 and S10L12 at the ratio of 1/1 (v/v) in ethanol (0.25 mg, 0.25 mg/ 10 μL) by
the ethanol injection method and heat treatment at 90 °C for 1 h.15 Subsequently,
S22L16 planar sheets dispersion (150 μL) were added to the round-bottom flask
dispersion (50 μL) and heated at 90 °C for 1 h. TEM images of molecular assemblies
were obtained (Fig. S4).
Figure S5. TEM images (negative staining with uranyl acetate) of molecular
assemblies from mixtures of S25L12 and S10L12 three-way tubes and S22L16
planar sheets. The assemblies were prepared in 10 mM Tris-HCl buffer (pH 7.4)
8