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  • Review Article
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Synthesis and applications of mirror-image proteins

Nature Reviews Chemistry volume 7, pages 383–404 (2023)Cite this article

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Abstract

The homochirality of biomolecules in nature, such as DNA, RNA, peptides and proteins, has played a critical role in establishing and sustaining life on Earth. This chiral bias has also given synthetic chemists the opportunity to generate molecules with inverted chirality, unlocking valuable new properties and applications. Advances in the field of chemical protein synthesis have underpinned the generation of numerous ‘mirror-image’ proteins (those comprised entirely of d-amino acids instead of canonical l-amino acids), which cannot be accessed using recombinant expression technologies. This Review seeks to highlight recent work on synthetic mirror-image proteins, with a focus on modern synthetic strategies that have been leveraged to access these complex biomolecules as well as their applications in protein crystallography, drug discovery and the creation of mirror-image life.

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Fig. 1: Synthetic strategies for mirror-image proteins.
Fig. 2: The racemic and quasi-racemic protein X-ray crystallography workflow and application for accelerating crystallization of synthetic proteins.
Fig. 3: Mirror-image phage display: a powerful platform for generating d-peptide/protein binders against a target of interest.
Fig. 4: Additional mirror-image screening techniques.
Fig. 5: The mirror-image central dogma with synthetic schemes of key mirror-image d-polymerases, d-African swine fever virus polymerase X and mutant of Sulfolobus solfataricus d-Dpo4.
Fig. 6: Construction of critical componentry required for the generation of mirror-image life.
Fig. 7: Synthesis of mirror-image mutant split-protein T7 RNA polymerase and mirror-image DNA ligase.
Fig. 8: Approaches to creating a mirror-image artificial cell.

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Acknowledgements

The authors acknowledge the Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science (CE200100012) and the NHMRC Investigator Grant APP1174941 (to R.J.P.) for funding and the Research Training Scholarship Program for PhD support (to A.S.M., L.K. and J.W.C.M.).

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  1. School of Chemistry, The University of Sydney, Sydney, New South Wales, Australia

    Katriona Harrison, Angus S. Mackay, Lucas Kambanis, Joshua W. C. Maxwell & Richard J. Payne

  2. Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, The University of Sydney, Sydney, New South Wales, Australia

    Katriona Harrison, Angus S. Mackay, Lucas Kambanis, Joshua W. C. Maxwell & Richard J. Payne

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Contributions

K.H. researched data for the article and contributed to writing, preparation of figures and reviewing and editing the manuscript. A.S.M. contributed to writing, preparation of figures and reviewing and editing the manuscript. L.K. and J.W.C.M contributed to preparation of the figures and revised the manuscript. R.J.P. revised the manuscript and conceived the overall direction of the manuscript. All authors have given approval to the final version of the manuscript.

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Harrison, K., Mackay, A.S., Kambanis, L. et al. Synthesis and applications of mirror-image proteins. Nat Rev Chem 7, 383–404 (2023). https://doi.org/10.1038/s41570-023-00493-y

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