The peptides were synthesized on
an automatic peptide synthesizer (Syro I, Biotage) by using a Rink-amide
resin and Fmoc chemistry. The Fmoc deprotection was carried out with
25% piperidine in DMF/NMP (70:30, v/v) for 3 min and 12.5% piperidine
in DMF/NMP (70:30, v/v) for 12 min. The couplings were accomplished
with the mixture Fmoc-AA-OH/HOBt/HBTU/DIPEA (5:5:4.8:10 equiv) for
2 × 40 min. N-terminal acetylation was performed manually with
acetic anhydride/DIPEA (10:10 equiv) in DMF for 30 min. The peptides
were cleaved from the resin with TFA/H2O/TIA/EDT/TIS (90:1:3:3:3; Vtot = 1 mL) for about 3 h, precipitated by ice-cold
diethyl ether, and recovered by centrifugation at 4 °C for 5
min. The homogeneity and identity of the lyophilized peptides were
assessed by analytical HPLC (Thermo Fisher Scientific) and MALDI-TOF-MS
(Bruker Daltonics) (Figures S20 and S21 and Table S7 ).
an automatic peptide synthesizer (Syro I, Biotage) by using a Rink-amide
resin and Fmoc chemistry. The Fmoc deprotection was carried out with
25% piperidine in DMF/NMP (70:30, v/v) for 3 min and 12.5% piperidine
in DMF/NMP (70:30, v/v) for 12 min. The couplings were accomplished
with the mixture Fmoc-AA-OH/HOBt/HBTU/DIPEA (5:5:4.8:10 equiv) for
2 × 40 min. N-terminal acetylation was performed manually with
acetic anhydride/DIPEA (10:10 equiv) in DMF for 30 min. The peptides
were cleaved from the resin with TFA/H2O/TIA/EDT/TIS (90:1:3:3:3; Vtot = 1 mL) for about 3 h, precipitated by ice-cold
diethyl ether, and recovered by centrifugation at 4 °C for 5
min. The homogeneity and identity of the lyophilized peptides were
assessed by analytical HPLC (Thermo Fisher Scientific) and MALDI-TOF-MS
(Bruker Daltonics) (
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