Intein method

EPL extends the applicability of native chemical ligation to recombinantly produced proteins using the mutant mini intein vector system. Proteins being expressed using this method will... 

M. Q. Xu, H. Paulus, and S. Chong, Fusions to self-splidng inteins for protein purification, Methods Enzymol. 2000, 326, 376-418. 

Lue, R. Y, Zhu, Q., Li, D. (2004). Site-specific peptide immobilization strategies for the rapid detection of kinase activity on microarrays. Site-specific immobilization of biotinylated proteins for protein microarray analysis. Enzymatic profiling system in a small-molecule microarray. Intein-mediated biotinylation of proteins and its application in a protein microarray. Developing site-specific immobilization strategies of peptides in a microarray. Methods Mol. Biol. 278, 191-204. 

In contrast to the relatively large number of reactions that target the N-terminus, only one generally effective method for C-terminal modification is currently available. This can be achieved through the use of intein-based methods to produce C-terminal thioesters, which can then be modified through NCLs with functionalized cysteines 38, Fig. 10.3-14 [49]. A more thorough description of the scope of this convenient method appears somewhere else in this book. 

Thermophilic inteins were among the first inteins discovered and played a key role in establishing protein splicing as a fundamental method of protein biosynthesis. The proof that inteins were spliced from precursor proteins rather than from precursor RNAs and the mechanism of protein splicing were initially demonstrated using archaeal inteins. Since their discovery in 1990, inteins have been harnessed to perform numerous protein engineering processes. 

---