Expressed Protein Ligation without Intein
نویسندگان
چکیده
منابع مشابه
Expressed protein ligation
The introduction of noncanonical amino acids and biophysical probes into peptides and proteins, and total or segmental isotopic labelling has the potential to greatly aid thedeterminationofprotein structure, functionandprotein– protein interactions. To obtain a peptide as large as possible by solid-phase peptide synthesis, native chemical ligation was introduced to enable synthesis of proteins ...
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The introduction of non-natural amino acid residues or modules into proteins provides a new means to explore the basis for conformational stability, folding/unfolding behavior, or biological function. We exploited intein-mediated protein ligation to produce a semisynthetic ribonuclease A. Of the 124 residues of RNase A, residues 1-94 were linked to an intein. After expression of the fusion prot...
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UNLABELLED Protein splicing in trans by split inteins has become a useful tool for protein engineering in vivo and in vitro. Inteins require Cys, Ser or Thr at the first residue of the C-terminal flanking sequence because a thiol or hydroxyl group in the side chains is a nucleophile indispensable for the trans-esterification step during protein splicing. Newly-identified distinct sequences with...
متن کاملSelenocysteine in native chemical ligation and expressed protein ligation.
L-Selenocysteine (Sec or U) has been called the “21st amino acid”.1 Like the twenty common amino acids, selenocysteine is inserted during the translation of mRNA and has its own tRNASec and codon, UGA. This codon also serves as the opal stop codon. Decoding a UGA codon as one for selenocysteine requires a special structure in the 3′ untranslated region of the mRNA called a selenocysteine insert...
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Chemically modified proteins are invaluable tools for studying the molecular details of biological processes, and they also hold great potential as new therapeutic agents. Several methods have been developed for the site-specific modification of proteins, one of the most widely used being expressed protein ligation (EPL) in which a recombinant α-thioester is ligated to an N-terminal Cys-contain...
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ژورنال
عنوان ژورنال: Journal of the American Chemical Society
سال: 2020
ISSN: 0002-7863,1520-5126
DOI: 10.1021/jacs.0c00252