Synthetic peptides are important research tools across scientific fields, but obtaining these materials can be time consuming, as coupling of each amino acid is typically conducted for an hour or more. Two papers now provide a serious shake-up of these reaction conditions. Simon et al. start by revisiting the basic kinetics of each step, confirming via inline UV detection of a released FMOC protecting group that reactions are complete in seconds. Unsurprisingly, these rates are faster at elevated temperatures, but heating stored amino acid solutions can lead to degradation. The authors therefore constructed reaction vessels that use flow-based coupling, deprotection and washing steps to decrease reaction times, incorporating a heat exchanger to rapidly heat materials immediately prior to and during synthesis. The final optimized protocol takes less than two minutes to add each amino acid, with purities of diverse peptides in line with material made by standard practices. Mong et al. leverage this technology to generate peptide fragments of two model proteins, DAPRin pE59 and barnase, that are then coupled via native chemical ligation. Because the synthesis is so fast, the authors were able to identify and resolve numerous pitfalls typical of polypeptide synthesis, such as deletion products, premature chain termination and aspartimide formation, and provide guidelines for handling these problems. This new methodology should dramatically increase the availability of peptides and proteins for basic and applied studies.
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Goodman, C. A synthetic step change. Nat Chem Biol 10, 324 (2014). https://doi.org/10.1038/nchembio.1514
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DOI: https://doi.org/10.1038/nchembio.1514
