ArticleThe Journal of biological chemistry2013
Faster protein splicing with the Nostoc punctiforme DnaE intein using non-native extein residues.
Article in The Journal of biological chemistry, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 53 papers.
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Who cites it
53 citing papers in PubMed, 101 citations in OpenAlex.
- De Novo Discovery of Bioactive Cyclic Peptides via Cellular Selection of Genetically Encoded Libraries.Chembiochem : a European journal of chemical biology · 2026Review
- Split Intein-Mediated Engineering of Bacteriocins Through Backbone Circularization, Modular Assembly of Chimeras, and Inducible Circularization.Biomolecules · 2026Article
- Generating Ultra-Fast Protein trans-Splicing of a Cysteine-Less and Semisynthetic Split Intein for Chemical Protein Labeling.Chembiochem : a European journal of chemical biology · 2026Article
- An enhanced split intein-mediated ligation (SIML) platform for rapid discovery and functional screening of circular bacteriocins.Frontiers in bioengineering and biotechnology · 2026Article
- Three-Segment Protein Labeling Using a Highly Efficient and Cysteine-Less Split Intein Identified with Computational Prediction of Aggregation Properties.Angewandte Chemie (International ed. in English) · 2025Article
- Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression.Nature communications · 2025Article
- Efficient genetic code expansion without host genome modifications.Nature biotechnology · 2025Article
- Intracellular protein editing enables incorporation of noncanonical residues in endogenous proteins.Science (New York, N.Y.) · 2025Article
- Optogenetics with Atomic Precision─A Comprehensive Review of Optical Control of Protein Function through Genetic Code Expansion.Chemical reviews · 2025Review
- Conditional Split Inteins: Adaptable Tools for Programming Protein Functions.International journal of molecular sciences · 2025Review
- Ligation of multiple protein domains using orthogonal inteins with non-native splice junctions.Protein science : a publication of the Protein Society · 2024Article
- Hox gene-specific cellular targeting using split intein Trojan exons.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Protein trans-splicing: optimization of intein-mediated GFP assembly as a model for the development of gene therapy.Frontiers in bioengineering and biotechnology · 2024Article
- Efficient splicing of the CPE intein derived from directed evolution of theActa biochimica et biophysica Sinica · 2023Article
- Extein residues regulate the catalytic function of Spl DnaX intein enzyme by restricting the near-attack conformations of the active-site residues.Protein science : a publication of the Protein Society · 2023Article
- Nature-inspired protein ligation and its applications.Nature reviews. Chemistry · 2023Review
- A Conserved Histidine Residue Drives Extein Dependence in an Enhanced Atypically Split Intein.Journal of the American Chemical Society · 2022Article
- Full-length ATP7B reconstituted through proteinMolecular therapy. Methods & clinical development · 2022Article
- Spatio-Temporal Photoactivation of Cytotoxic Proteins.Chembiochem : a European journal of chemical biology · 2022Article
- SufB intein splicing in Mycobacterium tuberculosis is influenced by two remote conserved N-extein histidines.Bioscience reports · 2022Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Inteins are naturally occurring intervening sequences that catalyze a protein splicing reaction resulting in intein excision and concatenation of the flanking polypeptides (exteins) with a native peptide bond. Inteins display a diversity of catalytic mechanisms within a highly conserved fold that is shared with hedgehog autoprocessing proteins. The unusual chemistry of inteins has afforded powerful biotechnology tools for controlling enzyme function upon splicing and allowing peptides of different origins to be coupled in a specific, time-defined manner. The extein sequences immediately flanking the intein affect splicing and can be defined as the intein substrate. Because of the enormous potential complexity of all possible flanking sequences, studying intein substrate specificity has been difficult. Therefore, we developed a genetic selection for splicing-dependent kanamycin resistance with no significant bias when six amino acids that immediately flanked the intein insertion site were randomized. We applied this selection to examine the sequence space of residues flanking the Nostoc punctiforme Npu DnaE intein and found that this intein efficiently splices a much wider range of sequences than previously thought, with little N-extein specificity and only two important C-extein positions. The novel selected extein sequences were sufficient to promote splicing in three unrelated proteins, confirming the generalizable nature of the specificity data and defining new potential insertion sites for any target. Kinetic analysis showed splicing rates with the selected exteins that were as fast or faster than the native extein, refuting past assumptions that the naturally selected flanking extein sequences are optimal for splicing.
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