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Titlebook: Expressed Protein Ligation; Methods and Protocol Miquel Vila-Perelló Book 2020 Springer Science+Business Media, LLC, part of Springer Natur

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Patricia M. Davies MCSP Dip. Phys. Ed. for EPL modification of both yeast surface displayed and secreted proteins with bioorthogonal chemical groups are described. These methods allow for the site-specific modification of intein-fused proteins produced in yeast.
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Methods to Study the Structure and Catalytic Activity of ,-Splicing Inteins,ction of inteins can have significant impact for biotechnology applications. Here, we provide biochemical methods to study splicing activity and NMR methods to study intein structure and the catalytic mechanism.
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Peptide Hydrazides as Thioester Equivalents for the Chemical Synthesis of Proteins,peptide hydrazides can be rapidly converted to peptide thioesters, which then selectively react with recombinant protein containing an N-terminal cysteine (Cys) to form a native peptide bond, thereby linking the two peptide segments without isolating any intermediates.
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Site-Directed Modification of Yeast-Produced Proteins Using Expressed Protein Ligation, for EPL modification of both yeast surface displayed and secreted proteins with bioorthogonal chemical groups are described. These methods allow for the site-specific modification of intein-fused proteins produced in yeast.
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,Die innere Uhr - Was lässt uns ticken?,tion of nature’s inteins which are protein modules that catalyze protein splicing. This chapter discusses the basic principles of expressed protein ligation and recent advances and applications in this protein semisynthesis field. Comparative strengths and weaknesses of the method and future challenges are highlighted.
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https://doi.org/10.1007/978-3-531-90468-9mal ligation sites are discussed. In addition, several methods used to generate the reactive fragments required for EPL are highlighted in practical details. Finally, strategies that one can implement to achieve efficient ligation reactions are presented.
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