ArticleBiotechnology and bioengineering2013
A redox trap to augment the intein toolbox.
Article in Biotechnology and bioengineering, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 20 citations in OpenAlex.
- Backbone Cyclization of Flavin Mononucleotide-Based Fluorescent Protein Increases Fluorescence and Stability.Journal of microbiology and biotechnology · 2023Article
- Conditional Alternative Protein Splicing Promoted by Inteins fromBiochemistry · 2022Article
- Inteins in Science: Evolution to Application.Microorganisms · 2020Review
- Methods to Study the Structure and Catalytic Activity of cis-Splicing Inteins.Methods in molecular biology (Clifton, N.J.) · 2020Article
- Structure of an engineered intein reveals thiazoline ring and provides mechanistic insight.Biotechnology and bioengineering · 2019Article
- Biotechnological Applications of Protein Splicing.Current protein & peptide science · 2019Review
- Biotechnology of extremely thermophilic archaea.FEMS microbiology reviews · 2018Review
- Exploring Intein Inhibition by Platinum Compounds as an Antimicrobial Strategy.The Journal of biological chemistry · 2016Article
- Intein applications: from protein purification and labeling to metabolic control methods.The Journal of biological chemistry · 2014Review
- Enigmatic distribution, evolution, and function of inteins.The Journal of biological chemistry · 2014Review
- Article
- Internal disulfide bond acts as a switch for intein activity.Biochemistry · 2013Article
- A conserved threonine spring-loads precursor for intein splicing.Protein science : a publication of the Protein Society · 2013Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
The unregulated activity of inteins during expression and consequent side reactions during work-up limits their widespread use in biotechnology and chemical biology. Therefore, we exploited a mechanism-based approach to regulate intein autocatalysis for biotechnological application. The system, inspired by our previous structural studies, is based on reversible trapping of the intein's catalytic cysteine residue through a disulfide bond. Using standard mutagenesis, the disulfide trap can be implemented to impart redox control over different inteins and for a variety of applications both in vitro and in Escherichia coli. Thereby, we first enhanced the output for bioconjugation in intein-mediated protein ligation, also referred to as expressed protein ligation, where precursor recovery and product yield were augmented fourfold to sixfold. Second, in bioseparation experiments, the redox trap boosted precursor recovery and product yield twofold. Finally, the disulfide-trap intein technology stimulated development of a novel bacterial redox sensor. This sensor reliably identified hyperoxic E. coli harboring mutations that disrupt the reductive pathways for thioredoxin and glutathione, against a background of wild-type cells.
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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.