Supporting Information

for Adv. Healthcare Mater., DOI: 10.1002/adhm.202101770

Antimicrobial polypeptides capable of membrane

translocation for treatment of MRSA wound infection in vivo
Shengcai Yang, Yanming Wang, Jason Tan, Jye Yng Teo, Ko Hui Tan, and Yi Yan Yang*

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Supporting Information

Antimicrobial polypeptides capable of membrane translocation for treatment of MRSA

wound infection in vivo

Shengcai Yang, Yanming Wang, Jason Tan, Jye Yng Teo, Ko Hui Tan, and Yi Yan Yang*

Institute of Bioengineering and Bioimaging, 31 Biopolis Way, Singapore 138669, Singapore

E-mail: yyyang@ibb.a-star.edu.sg

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Figure S1. 1H NMR spectra of (A) PLL35-Gua(Boc) in CDCl3 and (B) PLL35-Gua in DMSO-
d6.

Figure S2. GPC traces of (A) PLL22-Gua and (B) PLL35-Gua.

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Figure S3. 1H NMR spectra of (A) PLL22-Gua(Boc)(64%) in CDCl3 and (B) PLL22-
Gua(64%) in DMSO-d6.

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Figure S4. 1H NMR spectra of (A) PLL35-Gua(Boc)-VitE in CDCl3 and (B) PLL35-Gua-VitE
in DMSO-d6.

Figure S5. Critical micelle concentration (CMC) of (A) PLL22-Gua-VitE and (B) PLL35-Gua-
VitE in PBS.

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 Scheme S1. Synthesis of p(lys-r-leu).

Figure S6. 1H NMR spectra of (A) p(lys(Z)-r-leu) in TFA-d and (B) p(lys-r-leu) in D2O.

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 Figure S7. GPC traces of (A) p(lys-r-leu) and (B) p(lys(Gua)-r-leu).

Figure S8. 1H NMR spectra of (A) p(lys(Gua(Boc))-r-leu) in CDCl3 and (B) p(lys(Gua)-r-
leu) in D2O.

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Figure S9. Hemolytic activity of PLL, PLL-Gua, PLL-Gua-VitE, p(lys-r-leu) and p(lys(Gua)-
r-leu). The experiment was performed in triplicates, and the results were expressed as mean
hemolysis (%)  standard deviations shown by the error bars.

100
P L L 2 2-G u a

80 PLL22
C e ll V ia b ilit y ( % )

60

40

20

0
h
h

h
4
1

4
2

2
@

@
9

9
2

3
2
9

9
9

2
L

K
E
H

Figure S10. Viability of L929 or HEK293 cells after 24 h of incubation with PLL22 and
PLL22-Gua against at their respective MBC against MRSA.

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 Figure S11. Killing kinetics of vancomycin against MRSA within 3.0 h.

Figure S12. Photo of MRSA-infected wounds. For the establishment of wound infection
model, hair from the back of anaesthetized mice was removed and the exposed skin was
cleansed with 10% povidone-iodine solution and 75% ethanol. Three parallel wounds, with
about 1 cm length, were then created and infected with logarithmic growth of MRSA (1×10 10
CFU/mL, 60 µL) for 1 h prior to treatment.

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Figure S13. Photos of control mice (right panel) and PLL22-Gua-treated mice (left panel).
Treatment with 200 mg/kg of polymer did not affect the normal growth of fur and the body
weight of mice from both groups showed no significant difference.

Figure S14. H&E staining of normal tissues from control mice and PLL22-Gua treated mice.
The treatment with the polypeptide did not cause any damage to healthy tissues.

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