Expanding the chemical toolbox for the synthesis of large and uniquely modified proteins (original) (raw)

Abstract

O ver 100 years have passed since the first attempts to form amide bonds using simple starting materials. Despite this, great efforts are still being invested to develop new methods of efficiently forming amide bonds at low cost using environmentally friendly reagents 1. Synthesis of short peptide sequences (20-40 amino acids (AA)) is considered a relatively straightforward process with some exceptions such as amyloid β-peptides, transmembrane peptides and amphiphilic peptides. These difficult peptide sequences require special coupling reagents and synthetic strategies to achieve efficient synthesis. The relative ease by which essentially any short tailored peptide can be synthesized by applying solid-phase peptide synthesis (SPPS) has arisen from the continuous development of a wide variety of excellent coupling reagents, resins with better physical properties, novel linkers and orthogonal protecting groups 2. Improvement in chromatographic methods and analytical instrumentation for purification and characterization has also simplified the process of synthesizing peptides. The high efficiency of peptide synthesis coupled with automation has greatly expanded the use of peptides in several research areas, such as nanotechnology, drug discovery and total chemical synthesis of proteins. Chemical protein synthesis, which relies on chemoselective peptide ligation methods, such as native chemical ligation (NCL) 3 , offers complete control of the atomic composition of the polypeptide sequence, making it advantageous over molecular biology approaches in some areas of research (Fig. 1) 4. For example, it enables the incorporation of post-translational modifications (PTMs) into proteins with high homogeneity and in usable quantities, which can enable the role of these modifications on the protein structure and function to be examined. This method for the production of posttranslationally modified proteins has generated immense interests among many groups who have prepared, for example, glycosylated, phosphorylated and ubiquitinated proteins that have enabled otherwise difficult studies 5. Another exciting aspect of chemical protein synthesis is the ability to prepare mirror-image proteins, which are increasingly finding impressive applications in drug discovery 6 and protein crystallography 7 .

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