ArticleJournal of molecular biology2007
Crystallographic and mutational studies of Mycobacterium tuberculosis recA mini-inteins suggest a pivotal role for a highly conserved aspartate residue.
Article in Journal of molecular biology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers.
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Who cites it
46 citing papers in PubMed, 87 citations in OpenAlex.
- Regulation of N-terminal cleavage and splicing in mini-Ter DnaE-3 intein by E23G and R50Q mutations.Biotechnology letters · 2026Article
- Designing a Whole-Cell Biosensor for Detection of Toxic Metals Using Intein Splicing Inhibition ofACS omega · 2026Article
- A Unique THN Motif Is Critical for Enabling Efficient C-Terminal Traceless Cleavage.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Engineering of extracellular vesicles for efficient intracellular delivery of multimodal therapeutics including genome editors.Nature communications · 2025Article
- Article
- 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
- The Evolution of Intein-Based Affinity Methods as Reflected in 30 years of Patent History.Frontiers in molecular biosciences · 2022Review
- The Effects of Self-cleavage Intein-ELK16 Tag in the Transcript Steric Hindrance of IFN.Indian journal of clinical biochemistry : IJCB · 2021Article
- Intein Inhibitors as Novel Antimicrobials: Protein Splicing in Human Pathogens, Screening Methods, and Off-Target Considerations.Frontiers in molecular biosciences · 2021Review
- Structural, Dynamic, and Functional Characterization of a DnaX Mini-intein Derived fromBiochemistry · 2020Article
- Inteins in Science: Evolution to Application.Microorganisms · 2020Review
- Allosteric Influence of Extremophile Hairpin Motif Mutations on the Protein Splicing Activity of a Hyperthermophilic Intein.Biochemistry · 2020Article
- Spliceosomal Prp8 intein at the crossroads of protein and RNA splicing.PLoS biology · 2019Article
- Structure of an engineered intein reveals thiazoline ring and provides mechanistic insight.Biotechnology and bioengineering · 2019Article
- Cisplatin protects mice from challenge ofEmerging microbes & infections · 2019Article
- Mycobacterial DnaB helicase intein as oxidative stress sensor.Nature communications · 2018Article
- Exploring Intein Inhibition by Platinum Compounds as an Antimicrobial Strategy.The Journal of biological chemistry · 2016Article
- Review
- Backbone assignments of mini-RecA intein with short native exteins and an active N-terminal catalytic cysteine.Biomolecular NMR assignments · 2015Article
- Dynamics differentiate between active and inactive inteins.European journal of medicinal chemistry · 2015Article
Corrections and comments
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Authors and funding
6 authors at 4 institutions in 1 country.
Funding
Abstract
The 440 amino acid Mtu recA intein consists of independent protein-splicing and endonuclease domains. Previously, removal of the central endonuclease domain of the intein, and selection for function, generated a 168 residue mini-intein, DeltaI-SM, that had splicing activity similar to that of the full-length, wild-type protein. A D422G mutation (DeltaI-CM) increased C-terminal cleavage activity. Using the DeltaI-SM mini-intein structure (presented here) as a guide, we previously generated a highly active 139 residue mini-intein, DeltaDeltaI(hh)-SM, by replacing 36 amino acid residues in the residual endonuclease loop with a seven-residue beta-turn from the autoprocessing domain of Hedgehog protein. The three-dimensional structures of DeltaI-SM, DeltaDeltaI(hh)-SM, and two variants, DeltaDeltaI(hh)-CM and DeltaDeltaI(hh), have been determined to evaluate the effects of the minimization on intein integrity and to investigate the structural and functional consequences of the D422G mutation. These structural studies show that Asp422 is capable of interacting with both the N and C termini. These interactions are lacking in the CM variant, but are replaced by contacts with water molecules. Accordingly, additional mutagenesis of residue 422, combined with mutations that isolate N-terminal and C-terminal cleavage, showed that the side-chain of Asp422 plays a role in both N and C-terminal cleavage, thereby suggesting that this highly conserved residue regulates the balance between the two reactions.
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