摘要
我们设计开发了一种新的光交联探针,此探针运用二吖丙啶作为光交联基团捕捉蛋白-蛋白相互作用,运用生物正交反应分离纯化交联后产生的多肽,随后利用质谱对富集的多肽进行检测。新的光交联探针能够通过遗传密码子拓展技术定点插入到目标蛋白中。我们在大肠杆菌抗酸伴侣蛋白HdeA上展示了新型探针的流程,然后在模型蛋白上对交联的肽段进行了富集和鉴定。
We developed a genetivally and site-specifically encoded photo-cross-linker that makes use of diazirine as the photo-cross-linking group and exploits click chemistry to enrich the photo-cross-linked peptides for subsequent mass spectroscopy identification.We used the acid-protection chaperone,HdeA,in E.coli.to demonstrate the workflow of this new unnatural amino acid.
引文
[1]Zhang Meng,Lin Shixian,Song Xiwen,et al.Nature Chemical Biology,2011,7(10):671-677.
[2]Yihui Chen,Yalin Wu,Peter Henklein,et al.Chem.Eur.J.2010,16(16):7389-94.