Accelerated Synthesis of Modified Proteins via Novel Ligation Technologies

Abstract

Over the past decade there has been a renaissance in the use of large polypeptides and proteins as therapeutic agents. This has led to significant demand for technologies to rapidly and efficiently access these biomolecules, especially those bearing tailor-made modifications to maximize specificity and activity or to probe biological function (e.g. through incorporation of post-translational modifications, fluorophores and/or imaging reagents).1

We have recently developed a number of technologies that enable rapid access to proteins through chemical synthesis. This includes the development of methods for the one-pot synthesis of proteins.2 The utility of this methodology will be highlighted by describing the synthesis and biological activity of a library of post-translationally modified proteins.3 A more recently developed methodology is the diselenide-selenoester ligation reaction.4,5 This technology enables the fusion of peptide fragments with unprecedented kinetics and can be coupled with in situ deselenization chemistry to afford native polypeptides and proteins.5,6 The power of the methodology will be highlighted in this talk by discussing the use of the new technology in: 1) the rapid generation of a library of post-translationally modified viral proteins that selectively bind inflammatory human chemokines7 and 2) the total synthesis of a library of modified anti-thrombotic proteins, each of which could be assembled, purified and quantified for a bioassay within a few hours.8

 

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