Solid-phase peptide synthesis: from standard procedures to the synthesis of difficult sequences
Solid-phase peptide synthesis: from standard procedures to the synthesis of difficult sequences
复制标题
DOI：
10.1038/nprot.2007.454
复制
发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Bienert, Michael
中科院分区：
文献类型：
--
作者：
Coin, Irene;Beyermann, Michael;Bienert, Michael
该固相肽合成（SPPS）方案基于广泛使用的Fmoc/tBu策略、通过铵衍生的偶联试剂活化羧基和使用PEG改性的聚苯乙烯树脂。描述了一个标准的协议，这是成功地应用于我们的实验室合成促肾上腺皮质激素释放因子（CRF），>400 CRF类似物和无数的其他肽。41-mer肽CRF在类似于80个工作小时内获得。为了获得所谓的困难序列，必须应用特殊技术以减少生长肽链的聚集，这是肽化学合成失败的主要原因。显示了缩肽和假脯氨酸单元的示例性应用，用于合成极其困难的序列，WW结构域FBP 28的Asn（15）类似物，其不可能使用标准方案获得。
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.