Rapid preparation of stable isotope labeled peptides that bind to target proteins by a phage library system
Rapid preparation of stable isotope labeled peptides that bind to target proteins by a phage library system
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DOI：
10.1007/s10858-005-5054-0
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发表时间：
2006-01-01
影响因子：
2.7
通讯作者：
Shimada, I
中科院分区：
文献类型：
--
作者：
Mizukoshi, Y;Takahashi, H;Shimada, I
我们已经开发了一个系统，用于直接分离展示在噬菌体M13的主要外壳蛋白的N-末端上的外源肽。该系统中的噬菌体颗粒形成为野生型和修饰的外壳蛋白的混合物。将修饰的外壳蛋白的N-末端区段突变用于化学切割，以从主要外壳蛋白获得展示的肽。使用C-13、N-15标记的培养基，我们将稳定同位素C-13和/或N-15引入外壳蛋白。在选择噬菌体克隆后的几天内，可以记录来自噬菌体颗粒的裂解肽的NMR谱。
We have developed a system for directly isolating foreign peptides displayed on the N-terminus of the major coat protein of bacteriophage M13. The phage particle in this system is formed as a mixture of wild type and modified coat proteins. The N-terminal segment of the modified coat protein was mutated for chemical cleavage, in order to obtain the displayed peptide from the major coat protein. Using C-13, N-15- labeled medium, we introduced stable isotopes, C-13 and/or N-15, into the coat proteins. The NMR spectra for the cleaved peptides from the phage particles could be recorded within a few days after the selection of the phage clone.