University Of Illinois · Web view2018/08/01 · 13. 1H NMR spectrum of Anth-L-Glu-NCA in CDCl 3....
Transcript of University Of Illinois · Web view2018/08/01 · 13. 1H NMR spectrum of Anth-L-Glu-NCA in CDCl 3....
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Supporting Information
Manipulating the Membrane Penetration Mechanism of Helical Polypeptides via Aromatic Modification for Efficient Gene Delivery
Nan Zhenga,b,§, Ziyuan Songa,§, Jiandong Yang c,§, Yang Liua, Fangfang Lic, Jianjun Chenga,*, Lichen Yinc,*
a Department of Materials Science and Engineering, University of Illinois at Urbana–Champaign, 1304 W Green Street, Urbana, Illinois 61801, USA
b State Key Laboratory of Fine Chemicals, Department of Polymer Science and Engineering, School of Chemical Engineering, Dalian University of Technology, Dalian 116024, P. R. China
c Institute of Functional Nano & Soft Materials (FUNSOM), Jiangsu Key Laboratory for Carbon-Based Functional Materials & Devices, Collaborative Innovation Center of Suzhou Nano
Science and Technology, Soochow University, Suzhou 215123, P. R. China
* Corresponding author: E-mail: [email protected]; Phone: +86 0512-65882039 (L.Yin)
* Corresponding author: E-mail: [email protected]; Phone: +1 217-244-3924; Fax: 217-333-2736 (J. Cheng)
§ These authors contributed equally.
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Scheme S1. Synthetic routes of aromatic glutamate (A), aromatic glutamate based NCA monomers (B), and the L-Leu-NCA monomer (C).
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Table S1. Secondary conformational analysis of polypeptides
Name − [θ]222×10-3 (cm2 deg dmol-1) a) Helical content (%) b)
P0 30.0 84.7
P1 27.6 78.3
P2 24.6 70.8
P3 27.2 77.4
P4 22.6 65.6
P5 31.4 88.3
P6 32.9 92.0
P7 27.5 78.3
P8 19.9 58.7
a) The mean residue ellipticity [θ] was determined by following formula: Ellipticity ([θ] in cm2 deg dmol-1) = (millidegrees × mean residue weight) / (path length in mm × concentration of polypeptide in mg ml-1). b) The helical contents of the polypeptides were calculated by the following equation: helical content = (− [θ222] + 3000) / 39000.
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Fig. S1. CLSM images of HeLa cells following incubation with RhB-P0 and RhB-P3 at 37 °C or 4 °C for 2 h. Cell nuclei were stained with DAPI. Bar represents 20 μm.
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Fig. S2. Cell penetration levels of RhB-labeled polypeptides in HeLa cells in the presence of various endocytosis inhibitors including mβCD (A), chlorpromazine (B), and wortmannin (C) (n = 3).
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Fig. S3. DNA condensation by polypeptides at various polypeptide/DNA weight ratios as evaluated by the gel retardation assay. N represents naked DNA.
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Fig. S4. DNA condensation by polypeptides at different polypeptide/DNA weight ratios as determined by the EB exclusion assay (n = 3).
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Fig. S5. Particle size and zeta potential of polypeptide/DNA complexes at different polypeptide/DNA weight ratios.
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Fig. S6. Transfection efficiencies of P0 and P3 at various polypeptide/DNA weight ratios in B16F10 cells in the absence (A) or presence (B) of 10% serum (n = 3).
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Fig. S7. Cytotoxicity of polypeptide/DNA polyplexes following 24-h incubation in HeLa cells as determined by the MTT assay (n = 3).
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Fig. S8. Cytotoxicity of P0, P3, and PEI (25 kDa) at various concentrations in HeLa cells following 24-h incubation as determined by the MTT assay (n = 3).
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1H NMR spectra of new compounds.
Fig. S9. 1H NMR spectrum of Naph-L-Glu in DMSO-d6:DCl-D2O (9:1, v/v).
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Fig. S10. 1H NMR spectrum of Anth-L-Glu in DMSO-d6:DCl-D2O (9:1, v/v).
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Fig. S11. 1H NMR spectrum of B-L-Glu-NCA in CDCl3.
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Fig. S12. 1H NMR spectrum of Naph-L-Glu-NCA in CDCl3.
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Fig. S13. 1H NMR spectrum of Anth-L-Glu-NCA in CDCl3.
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Fig. S14. 1H NMR spectrum of L-Leu-NCA in CDCl3.
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Fig. S15. Representative 1H NMR spectrum of copolymer precursor for composition calculation (PALG-r-PABLG as an example, P5 precursor) in CDCl3:TFA-d (85:15, v/v). The block composition was calculated by the integration ratio of the α-protons in PALG residues (proton a) to the α-protons in PABLG residues (proton a’).
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Fig. S16. 1H NMR spectrum of P0 in TFA-d.
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Fig. S17. 1H NMR spectrum of P1 in TFA-d.
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Fig. S18. 1H NMR spectrum of P2 in TFA-d.
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Fig. S19. 1H NMR spectrum of P3 in TFA-d.
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Fig. S20. 1H NMR spectrum of P4 in TFA-d.
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Fig. S21. 1H NMR spectrum of P5 in TFA-d.
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Fig. S22. 1H NMR spectrum of P6 in TFA-d.
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Fig. S23. 1H NMR spectrum of P7 in TFA-d.
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Fig. S24. 1H NMR spectrum of P8 in TFA-d.
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