Synthesis of full length PB1-F2 influenza A virus proteins from 'Spanish flu' and 'bird flu'
Synthesis of full length PB1-F2 influenza A virus proteins from 'Spanish flu' and 'bird flu'
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DOI：
10.1002/psc.1031
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发表时间：
2008-08-01
影响因子：
2.1
通讯作者：
Schubert, Ulrich
中科院分区：
文献类型：
--
作者：
Roeder, Rene;Bruns, Karsten;Schubert, Ulrich
促凋亡甲型流感病毒PB 1-F2蛋白有助于病毒致病性，并存在于大多数人和禽类分离株中。先前的合成方案已被改进以提供由“西班牙流感”分离株编码的合成全长H1N1型PB 1-F2蛋白及其等同物。一种来自禽类宿主的蛋白质，代表高致病性H5 N1“禽流感”分离株，称为SF 2和BF 2。分别全长SF 2。已经通过标准固相肽合成方法或通过未保护的N-和C-末端肽片段的天然化学连接合成了BF 2的不同突变体和这些肽的许多片段。对于SF 2化学连接，利用分别位于天然序列的位置41和42的组氨酸和半胱氨酸残基。与标准SPPS延伸方法相比，提供了高效的SF 2合成。通过对SPPS方案进行特定修改，防止在位置23的天冬氨酸残基处形成副产物。而BF 2的天然序列则没有。含有半胱氨酸残基的BF 2（Y 42 C）和BF 2（S47 C）的两种不同突变体具有适当的半胱氨酸交换。除了全长分子外，还合成了天然序列的片段，用于将其物理特征与来自H1N1人分离株A/波多黎各/8/34（H1N1）的那些进行比较。通过质谱分析所有肽。1H-1 NMR光谱。SDS-PAGE。所述方案允许合成显著量的PB 1-F2及其相关肽。版权所有（C）2008欧洲肽协会和约翰威利父子。公司
The proapoptotic influenza A virus PB1-F2 protein contributes to viral pathogenicity and is present in most human and avian isolates. Previous synthetic protocols have been improved to provide a synthetic full length H1N1 type PB1-F2 protein that is encoded by the 'Spanish flu' isolate and an equivalent. protein from an avian host that is representative of a highly pathogenic H5N1 'bird flu' isolate, termed SF2and BF2. respectively. Full length SF2. different mutants of BF2 and a number of fragments of these peptides have been synthesized by either the standard solid-phase peptide synthesis method or by native chemical ligation of unprotected N- and C-terminal peptide fragments. For SF2 chemical ligation made use of the histidine and the cysteine residues located in positions 41 and 42 of the native sequence, respectively. to afford a highly efficient synthesis of SF2 compared to the standard SPPS elongation method. By-product formation at the aspartic acid residue in position 23 was prevented by specific modifications of the SPPS protocol. As the native sequence of BF2 does not. contain a cysteine residue two different mutants of BF2 (Y42C) and BF2 (S47C) with appropriate cysteine exchanges were produced. In addition to the full length molecules, fragments of the native sequences were synthesized for comparison of their physical characteristics with those from the H1N1 human isolate A/Puerto Rico/8/34 (H1N1). All peptides were analyzed by mass spectrometry. H-1 NMR spectroscopy. and SDS-PAGE. The protocols allow the synthesis of significant amounts of PB1-F2 and its related peptides. Copyright (C) 2008 European Peptide Society and John Wiley & Sons. Ltd.