ArticleJournal of molecular biology2011
Spontaneous proton transfer to a conserved intein residue determines on-pathway protein splicing.
Article in Journal of molecular biology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 27 citations in OpenAlex.
- A Genetically Encoded Homocysteine Precursor to Probe Protein Active Sites and to Addict Escherichia coli to a Noncanonical Amino Acid Directly Involved in Catalysis.Angewandte Chemie (International ed. in English) · 2025Article
- 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
- SufB intein splicing in Mycobacterium tuberculosis is influenced by two remote conserved N-extein histidines.Bioscience reports · 2022Article
- Unraveling the origin of interactions of hydroxychloroquine with the receptor-binding domain of SARS-CoV-2 in aqueous medium.Chemical physics letters · 2021Article
- Structure of an engineered intein reveals thiazoline ring and provides mechanistic insight.Biotechnology and bioengineering · 2019Article
- An Unprecedented Combination of Serine and Cysteine Nucleophiles in a Split Intein with an Atypical Split Site.The Journal of biological chemistry · 2015Article
- Förster resonance energy transfer-based cholesterolysis assay identifies a novel hedgehog inhibitor.Analytical biochemistry · 2015Article
- Review
- Backbone assignments of mini-RecA intein with short native exteins and an active N-terminal catalytic cysteine.Biomolecular NMR assignments · 2015Article
- SufB intein of Mycobacterium tuberculosis as a sensor for oxidative and nitrosative stresses.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Protein splicing: how inteins escape from precursor proteins.The Journal of biological chemistry · 2014Review
- Structural and dynamical features of inteins and implications on protein splicing.The Journal of biological chemistry · 2014Review
- Internal disulfide bond acts as a switch for intein activity.Biochemistry · 2013Article
- Oriented covalent immobilization of antibodies for measurement of intermolecular binding forces between zipper-like contact surfaces of split inteins.Analytical chemistry · 2013Article
- Recent progress in intein research: from mechanism to directed evolution and applications.Cellular and molecular life sciences : CMLS · 2013Review
- Structural and mutational studies of a hyperthermophilic intein from DNA polymerase II of Pyrococcus abyssi.The Journal of biological chemistry · 2011Article
- pK(a) coupling at the intein active site: implications for the coordination mechanism of protein splicing with a conserved aspartate.Journal of the American Chemical Society · 2011Article
Corrections and comments
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Authors and funding
6 authors at 3 institutions in 1 country.
Funding
Abstract
The discovery of inteins, which are protein-splicing elements, has stimulated interest for various applications in chemical biology, bioseparations, drug delivery, and sensor development. However, for inteins to effectively contribute to these applications, an increased mechanistic understanding of cleavage and splicing reactions is required. While the multistep chemical reaction that leads to splicing is often explored and utilized, it is not clear how the intein navigates through the reaction space. The sequence of reaction steps must progress in concert in order to yield efficient splicing while minimizing off-pathway cleavage reactions. In this study, we demonstrate that formation of a previously identified branched intermediate is the critical step for determining splicing over cleavage products. By combining experimental assays and quantum mechanical simulations, we identify the electrostatic interactions that are important to the dynamics of the reaction steps. We illustrate, via an animated simulation trajectory, a proton transfer from the first C-terminal extein residue to a conserved aspartate, which synchronizes the multistep enzymatic reaction that is key to splicing. This work provides new insights into the complex interplay between critical active-site residues in the protein splicing mechanism, thereby facilitating biotechnological application while shedding light on multistep enzyme activity.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.