Split Inteins介导的多肽链两端标记

Split Inteins Mediated Site-specific Dual-labeling of Proteins / Peptides

多肽链多位点特异性标记有助于了解蛋白质的结构与功能,特别是在蛋白质的动态构象研究方面.但是,现有的多肽链多位点特异性标记方法各有局限性,并且种类有限,所以有必要开发新的多肽链多位点特异性标记方法以满足研究需求.本文以Diub(ubiquitin dimer)蛋白为研究对象,借助S1和S11型2种不同的断裂蛋白质内含子(split inteins)的蛋白质反式剪接,将含有不同荧光基团的两种小肽成功剪接至靶蛋白的两端,最终达到对靶蛋白的末端标记目的.

Multi-site protein modifications and labeling can be very helpful for protein structural and functional studies,especially in protein dynamics studies. Novel methods are needed due to the limitation of current canonical multi-site protein modification methods. In this article,two novel split inteins（ S1 and S11） which have short complementary parts were successfully applied to splice the short peptides containing desired labeling to N- and C-terminus of the target by two steps of protein trans-splicing,respectively. Together,we have developed a protein / peptide dual-labeling technique,by which the target protein（ ubiquitin dimer） can be specifically labeled with a reasonable yield.