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Fine-tuning an engineered intein

Nature Biotechnology volume 17, pages 854–855 (1999)Cite this article

Although it is possible to introduce desirable properties and activities into proteins using rational design, subtle changes necessary to make an engineered product efficient and practical are often still beyond our predictive capacity1. In this issue, Belfort and colleagues2 describe an elegant mutational strategy to engineer an intein with improved features to serve as a tool for protein purification. Previously, this group created a mini-intein by removing an internal domain encoding an endonuclease activity required for horizontal transfer of the intein gene3. The endonuclease domain is distinct and separate in function, structure, and evolution from the protein splicing domain. The resulting mini-intein could splice, but not as efficiently as its unmodified progenitor. In the current work, this mini-intein was further engineered to both enhance the splicing efficiency and allow it to cleave solely at its C-terminal end.

Inteins are proteins whose genes are found only within the genes of other proteins. Although the intein and host gene are translated as a single polypeptide, the intein initiates an autocatalytic event to remove itself and join the flanking host segments with a new polypeptide bond. In addition to their protein splicing activity, many inteins also possess endonucleic activity that probably serves to propagate their genes to new integration sites. Thus, inteins are selfish genetic elements whose known functions only benefit themselves. Inteins are found in all three domains of life, yet in only 30 species. No host factors are needed for the protein splicing activity, and the process appears very efficient1. These novel attributes make inteins of potential utility in biotechnology, and several research groups are engineering them for particular applications. For example, intein-based systems have been used to ligate proteins. This enables the expression of toxic proteins5 (made in two inactive parts and joined after purification), the generation of novel protein combinations6, and the selective labeling of parts of proteins9 (especially useful for NMR studies of large proteins).

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Figure 1: An inside look at protein purification strategies using inteins.

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Authors and Affiliations

  1. the Molecular Genetics department at the Weizmann Institute of Science, Rehovot, 76100, Israel

    Gil Amitai & Shmuel Pietrokovski

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  1. Gil Amitai
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  2. Shmuel Pietrokovski
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Amitai, G., Pietrokovski, S. Fine-tuning an engineered intein. Nat Biotechnol 17, 854–855 (1999). https://doi.org/10.1038/12839

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