ArticleNucleic acids research2002
Intein-mediated purification of cytotoxic endonuclease I-TevI by insertional inactivation and pH-controllable splicing.
Article in Nucleic acids research, 2002. 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, 53 citations in OpenAlex.
- Mechanism-guided control of intracellular alkalinization and process pH for production of recombinant human B-type natriuretic peptide under the cSAT scheme in Escherichia coli.Microbial cell factories · 2026Article
- Intein-based thermoregulated meganucleases for containment of genetic material.Nucleic acids research · 2024Article
- A Photoresponsive Homing Endonuclease for Programmed DNA Cleavage.ACS synthetic biology · 2024Article
- A promiscuous split intein with expanded protein engineering applications.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Article
- Dynamics differentiate between active and inactive inteins.European journal of medicinal chemistry · 2015Article
- Article
- Protein trans-splicing of multiple atypical split inteins engineered from natural inteins.PloS one · 2013Article
- Assembly of protein building blocks using a short synthetic peptide.Bioconjugate chemistry · 2012Article
- Protein trans-splicing of an atypical split intein showing structural flexibility and cross-reactivity.PloS one · 2012Article
- Electronic structure of neighboring extein residue modulates intein C-terminal cleavage activity.Biophysical journal · 2011Article
- Spontaneous proton transfer to a conserved intein residue determines on-pathway protein splicing.Journal of molecular biology · 2011Article
- Redox-responsive zinc finger fidelity switch in homing endonuclease and intron promiscuity in oxidative stress.Current biology : CB · 2011Article
- A unified genetic, computational and experimental framework identifies functionally relevant residues of the homing endonuclease I-BmoI.Nucleic acids research · 2010Article
- Selection and structure of hyperactive inteins: peripheral changes relayed to the catalytic center.Journal of molecular biology · 2009Article
- Homing endonuclease I-TevIII: dimerization as a means to a double-strand break.Nucleic acids research · 2007Article
- Regulation of protein activity with small-molecule-controlled inteins.Protein science : a publication of the Protein Society · 2005Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
An intein-mediated approach was developed for expression and affinity purification of a protein that is lethal to Escherichia coli. The protein, I-TevI, is an intron-encoded endonuclease. The approach involved the insertional inactivation of I-TevI with a controllable mini-intein placed in front of a cysteine required for splicing (an I-TevI::intein fusion). The purification was facilitated by a chitin-binding domain inserted into the mini-intein. Affinity purification of the I-TevI::intein fusion precursor on a chitin column was followed by pH-controllable splicing to restore the structure and function of I-TevI. To study the impact of the insertion context on I-TevI inactivation, the chimeric intein was inserted independently in front of seven cysteines of I-TevI. One of the seven intein integrants yielded I-TevI of high activity. This technique is, in principle, generalizable to the expression and purification of other cytotoxic proteins and is amenable to scale-up.
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