Solid-phase peptide synthesis: from standard procedures to the synthesis of difficult sequences

被引:466
作者
Coin, Irene [1 ]
Beyermann, Michael [1 ]
Bienert, Michael [1 ]
机构
[1] Leibniz Inst Mol Pharmacol, Dept Peptide Chem & Biochem, D-13125 Berlin, Germany
关键词
D O I
10.1038/nprot.2007.454
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This protocol for solid-phase peptide synthesis (SPPS) is based on the widely used Fmoc/tBu strategy, activation of the carboxyl groups by aminium-derived coupling reagents and use of PEG-modified polystyrene resins. A standard protocol is described, which was successfully applied in our lab for the synthesis of the corticotropin-releasing factor (CRF), >400 CRF analogs and a countless number of other peptides. The 41-mer peptide CRF is obtained within similar to 80 working hours. To achieve the so-called difficult sequences, special techniques have to be applied in order to reduce aggregation of the growing peptide chain, which is the main cause of failure for peptide chemosynthesis. Exemplary application of depsipeptide and pseudoproline units is shown for synthesizing an extremely difficult sequence, the Asn(15) analog of the WW domain FBP28, which is impossible to obtain using the standard protocol.
引用
收藏
页码:3247 / 3256
页数:10
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