Abstract
Sufficient quantities of homogeneous samples of post-translationally modified proteins are often not readily available from biological sources to facilitate structure–function investigations. Native chemical ligation (NCL) is a convenient method for the production of biologically active proteins from smaller fragments. Such an approach allows protein modifications to be introduced in a controlled fashion into smaller peptide fragments which are amenable to total chemical synthesis. These fragments of defined sequence and structure can be elaborated to full-length proteins through NCL reactions with suitable components derived from bacterial origin. This report describes methods for the bacterial production of components for NCL and their use in typical reactions.
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The authors acknowledge financial support from The Royal Society and The Wellcome Trust.
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Richardson, J.P., Macmillan, D. (2011). Semi-synthesis of Glycoproteins from E. coliThrough Native Chemical Ligation. In: Evans, Jr., T., Xu, MQ. (eds) Heterologous Gene Expression in E.coli. Methods in Molecular Biology, vol 705. Humana Press. https://doi.org/10.1007/978-1-61737-967-3_9
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DOI: https://doi.org/10.1007/978-1-61737-967-3_9
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