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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Methods and Applications of Expressed Protein Ligation,N-cysteine containing synthetic peptides in a chemoselective ligation. The recombinant protein C-terminal thioesters are produced by exploiting the action of nature’s inteins which are protein modules that catalyze protein splicing. This chapter discusses the basic principles of expressed protein li

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Protein Splicing: From the Foundations to the Development of Biotechnological Applications,odifications, and biophysical probes into proteins of any size. This methodology has been developed based on the knowledge obtained from protein splicing. Protein splicing is a multistep biochemical reaction that includes the concomitant cleavage and formation of peptide bonds carried out by self-pr

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Identification, Characterization, and Optimization of Split Inteins,es, most notably protein ligation. The development of these approaches is dependent on the identification and/or design of split inteins with robust activity, stability, and solubility. Here, we describe two approaches to characterize and compare the activities of newly identified or engineered spli

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Methods to Study the Structure and Catalytic Activity of ,-Splicing Inteins,tein splicing has been studied by overexpression in . of intein fusion proteins with nonnative exteins. Inteins can be used to generate reactive α-thioesters, as well as proteins with N-terminal Cys residues, to facilitate expressed protein ligation. As such, a more detailed understanding of the fun

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