Abstract
Protein trans-splicing using split inteins is a powerful and convenient reaction to chemically modify recombinantly expressed proteins under mild conditions. In particular, semisynthetic protein trans-splicing with one intein fragment short enough to be accessible by solid-phase peptide synthesis can be used to transfer a short peptide segment with the desired synthetic moiety to the protein of interest. In this chapter, we provide detailed protocols for two such split intein systems. The M86 mutant of the Ssp DnaB intein and the MX1 mutant of the AceL-TerL intein are two highly engineered split inteins with very short N-terminal intein fragments of only 11 and 25 amino acids, respectively, and allow the efficient N-terminal labeling of proteins.
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Acknowledgements
We thank Xiang-Qin Liu (Dalhousie University, Canada) for collaboration on initial work on the M86 mutant and Shmuel Pietrokovski (Weizmann Institute, Israel) for collaboration on the AceL-TerL intein. We acknowledge funding by DFG (grants MO1073/3-2; SPP1623, MO1073/5-1, and Cells-in-Motion excellence cluster, EXC1003) and Fonds der Chemischen Industrie.
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Matern, J.C.J. et al. (2015). Ligation of Synthetic Peptides to Proteins Using Semisynthetic Protein trans-Splicing. In: Gautier, A., Hinner, M. (eds) Site-Specific Protein Labeling. Methods in Molecular Biology, vol 1266. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2272-7_9
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DOI: https://doi.org/10.1007/978-1-4939-2272-7_9
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