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Peptide Synthesis through Cell‐Free Expression of Fusion Proteins Incorporating Modified Amino Acids as Latent Cleavage Sites for Peptide Release
Author(s) -
Liutkus Mantas,
Fraser Samuel A.,
Caron Karine,
Stigers Dan J.,
Easton Christopher J.
Publication year - 2016
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201600091
Subject(s) - peptide , chemistry , peptide synthesis , biochemistry , amino acid , cholecystokinin , chemical ligation , cell free protein synthesis , peptide sequence , oligopeptide , protein biosynthesis , receptor , gene
Chlorinated analogues of Leu and Ile are incorporated during cell‐free expression of peptides fused to protein, by exploiting the promiscuity of the natural biosynthetic machinery. They then act as sites for clean and efficient release of the peptides simply by brief heat treatment. Dehydro analogues of Leu and Ile are similarly incorporated as latent sites for peptide release through treatment with iodine under cold conditions. These protocols complement enzyme‐catalyzed methods and have been used to prepare calcitonin, gastrin‐releasing peptide, cholecystokinin‐7, and prolactin‐releasing peptide prohormones, as well as analogues substituted with unusual amino acids, thus illustrating their practical utility as alternatives to more traditional chemical peptide synthesis.