Traceless protein semi-synthesis in cells using the promiscuous ultra-fast split intein NrdJ-1
Ye, X.; Sokol, J.; Kofoed, C.; Dheer, A.; Zhang, J.; Muir, T. W.
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Chemistry-driven approaches that allow proteins to be manipulated in ways not permitted by standard genetics are indispensable in the basic and applied biomedical sciences. Of the various platform technologies in common use, those that exploit intein-mediated protein splicing have proven especially powerful owing to compatibility with both in vitro and in vivo applications. Here, we provide detailed biochemical characterization of the split intein, NrdJ-1. We demonstrate that the rapid splicing kinetics associated with this system are largely independent of splice junction primary sequence context. The promiscuity and high efficiency of this split intein makes it well suited to the generation of chemically modified proteins in vitro and in living cells, as demonstrated by the traceless generation of semisynthetic chromatin harboring various post-translational modifications. These studies highlight the utility of NrdJ-1 for traceless protein editing and suggest it is a superior choice for many intracellular biochemical applications.
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