ArticleJournal of molecular biology2012
NMR and crystal structures of the Pyrococcus horikoshii RadA intein guide a strategy for engineering a highly efficient and promiscuous intein.
Article in Journal of molecular biology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 45 papers.
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Who cites it
45 citing papers in PubMed, 67 citations in OpenAlex.
- 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
- Monitoring Protein Stability In Vivo Using an Intein-Based Biosensor.Bio-protocol · 2025Article
- Controllable intein splicing and N-terminal cleavage at mesophilic temperatures.Frontiers in bioengineering and biotechnology · 2025Article
- Article
- An intein-based biosensor to measure protein stability in vivo.Protein science : a publication of the Protein Society · 2024Article
- 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
- Standard Intein Gene Expression Ramps (SIGER) for Protein-Independent Expression Control.ACS synthetic biology · 2023Article
- Nature-inspired protein ligation and its applications.Nature reviews. Chemistry · 2023Review
- Inteins-mechanism of protein splicing, emerging regulatory roles, and applications in protein engineering.Frontiers in microbiology · 2023Review
- Labeling Ebola Virus with a Self-Splicing Fluorescent Reporter.Microorganisms · 2022Article
- SufB intein splicing in Mycobacterium tuberculosis is influenced by two remote conserved N-extein histidines.Bioscience reports · 2022Article
- Protein Synthesis via Activated Cysteine-Directed Protein Ligation.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Mini-Intein Structures from Extremophiles Suggest a Strategy for Finding Novel Robust Inteins.Microorganisms · 2021Article
- An alternative domain-swapped structure of the Pyrococcus horikoshii PolII mini-intein.Scientific reports · 2021Article
- Structural, Dynamic, and Functional Characterization of a DnaX Mini-intein Derived fromBiochemistry · 2020Article
- Inteins in Science: Evolution to Application.Microorganisms · 2020Review
- The Convergence of the Hedgehog/Intein Fold in Different Protein Splicing Mechanisms.International journal of molecular sciences · 2020Article
- Protein Splicing Activity of theBiochemistry · 2020Article
- Allosteric Influence of Extremophile Hairpin Motif Mutations on the Protein Splicing Activity of a Hyperthermophilic Intein.Biochemistry · 2020Article
- The crystal structure of the naturally split gp41-1 intein guides the engineering of orthogonal split inteins from cis-splicing inteins.The FEBS journal · 2020Article
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
Abstract
In protein splicing, an intervening protein sequence (intein) in the host protein excises itself out and ligates two split host protein sequences (exteins) to produce a mature host protein. Inteins require the involvement for the splicing of the first residue of the extein that follows the intein (which is Cys, Ser, or Thr). Other extein residues near the splicing junctions could modulate splicing efficiency even when they are not directly involved in catalysis. Mutual interdependence between this molecular parasite (intein) and its host protein (exteins) is not beneficial for intein spread but could be advantageous for intein survival during evolution. Elucidating extein-intein dependency has increasingly become important since inteins are recognized as useful biotechnological tools for protein ligation. We determined the structures of one of inteins with high splicing efficiency, the RadA intein from Pyrococcus horikoshii (PhoRadA). The solution NMR structure and the crystal structures elucidated the structural basis for its high efficiency and directed our efforts of engineering that led to rational design of a functional minimized RadA intein. The crystal structure of the minimized RadA intein also revealed the precise interactions between N-extein and the intein. We systematically analyzed the effects at the -1 position of N-extein and were able to significantly improve the splicing efficiency of a less robust splicing variant by eliminating the unfavorable extein-intein interactions observed in the structure. This work provides an example of how unveiling structure-function relationships of inteins offer a promising way of improving their properties as better tools for protein engineering.
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