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Homing Endonucleases and Inteins pp 233–255Cite as

Biochemical Mechanisms of Intein-Mediated Protein Splicing

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Part of the book series: Nucleic Acids and Molecular Biology ((NUCLEIC,volume 16))

Abstract

This chapter discusses the mechanism of the self-catalyzed process by which inteins promote both their own excision from a host protein and the direct linkage of the flanking host protein segments, the N- and C-exteins, by a peptide bond. The majority of inteins have a nucleophilic amino acid at their N-terminus and asparagine at their C-terminus and are linked to a C-extein with an N-terminal nucleophilic amino acid. These canonical inteins promote protein splicing by a four-step mechanism of sequential acyl rearrangements. Non-canonical inteins, which lack either the N-terminal nucleophile or the C-terminal asparagine, promote protein splicing by a variant of this mechanism or promote protein cleavage rather than splicing. A remarkable feature of the protein splicing process is that it involves multiple steps that are chemically autonomous yet proceed in a highly coordinated manner without side reactions unless perturbed by mutation, unnatural exteins, or non-physiological conditions. The factors that may serve to integrate protein splicing into a system that ordinarily operates efficiently without side reactions are discussed.

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Author information

Authors and Affiliations

  1. Department of Chemistry, College of the Holy Cross, 1 College Street, Worcester, Massachusetts, 01610, USA

    Kenneth V. Mills

  2. Boston Biomedical Research Institute, 64 Grove Street, Watertown, Massachusetts, 02472, USA

    Henry Paulus

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  1. Kenneth V. Mills
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  2. Henry Paulus
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Editor information

Editors and Affiliations

  1. Wadsworth Center/NYSDOH, Center for Medical Science, 150 New Scotland Ave, Albany, NY, 12208, USA

    Marlene Belfort  & Victoria Derbyshire  & 

  2. Dept. of Chemical Engineering, Princeton University, A417 Engineering Quadrangle, Princeton, NJ, 08544, USA

    David W. Wood

  3. Fred Hutchinson Cancer Research, Center Institute, 1100 Fairview Ave. N. Mailstop A3-023, Seattle, WA, 98109, USA

    Barry L. Stoddard

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Mills, K.V., Paulus, H. (2005). Biochemical Mechanisms of Intein-Mediated Protein Splicing. In: Belfort, M., Wood, D.W., Stoddard, B.L., Derbyshire, V. (eds) Homing Endonucleases and Inteins. Nucleic Acids and Molecular Biology, vol 16. Springer, Berlin, Heidelberg. https://doi.org/10.1007/3-540-29474-0_14

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