反应详情

General Procedure for Peptide Synthesis. Tat49-57 (RKKRRQRRR; SEQ ID NO:28), truncated and alanine-substituted peptides derived from Tat49-57, Antennapedia43-58 (RQIKIWFQNRRMKWKKK SEQ ID NO:29), and homopolymers of arginine (R5-R9; SEQ ID NOS:1-5) and d-arginine (r5-r9) were prepared with an automated peptide synthesizer (ABI433) using standard solid-phase Fmoc chemistry (35) with HATU as the peptide coupling reagent. The fluorescein moiety was attached via a aminohexanoic acid spacer by treating a resin-bound peptide (1.0 mmol) with fluorescein isothiocyanate (1.0 mmol) and DIBA (5 mmol) in DMF (10 mL) for 12 h. Cleavage from the resin was achieved using 95:5 TFA/triisopropylsilane. Removal of the solvent in vacuo gave a crude oil which was triturated with cold ether. The crude mixture thus obtained was centrifuged, the ether was removed by decantation, and the resulting orange solid was purified by reverse-phase HPLC (H2O/CH3CN in 0.1% TFA). The products were isolated by lyophilization and characterized by electrospray mass spectrometry. Purity of the peptides was >95% as determined by analytical reverse-phase HPLC (H2O/CH3CN in 0.1% TFA).

1. customRemoval of the solvent in vacuo
2. customgave a crude oil which
3. customwas triturated with cold ether