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. 2022 Jan 10;61(2):e202110223.
doi: 10.1002/anie.202110223. Epub 2021 Dec 3.

Site-Selective Installation of Nϵ -Modified Sidechains into Peptide and Protein Scaffolds via Visible-Light-Mediated Desulfurative C-C Bond Formation

Rhys C Griffiths  1 , Frances R Smith  1 , Jed E Long  2 , Daniel Scott  3 , Huw E L Williams  2 , Neil J Oldham  1 , Robert Layfield  3 , Nicholas J Mitchell  1
Affiliations

Site-Selective Installation of Nϵ -Modified Sidechains into Peptide and Protein Scaffolds via Visible-Light-Mediated Desulfurative C-C Bond Formation

Rhys C Griffiths et al. Angew Chem Int Ed Engl. .

Abstract

Post-translational modifications (PTMs) enhance the repertoire of protein function and mediate or influence the activity of many cellular processes. The preparation of site-specifically and homogeneously modified proteins, to apply as tools to understand the biological role of PTMs, is a challenging task. Herein, we describe a visible-light-mediated desulfurative C(sp3 )-C(sp3 ) bond forming reaction that enables the site-selective installation of Nϵ -modified sidechains into peptides and proteins of interest. Rapid, operationally simple, and tolerant to ambient atmosphere, we demonstrate the installation of a range of lysine (Lys) PTMs into model peptide systems and showcase the potential of this technology by site-selectively installing an Nϵ Ac sidechain into recombinantly expressed ubiquitin (Ub).

Keywords: bioconjugation; cysteine; peptides; photochemistry; site selectivity.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Synthetic approaches to the site‐selective installation of Lys PTM mimics (A) and PTMs (B); N ϵ Ac shown. Our approach (C): site‐selective installation of N ϵ ‐modified sidechains via interception of visible‐light‐mediated desulfurization.
Figure 2
Figure 2
NMR spectra (CHα region) of N ϵ Ac sidechain installation using model peptides 4 a and 4 b to afford epimers 5 a (B) and 5 b (A); NMR spectra of 5 a synthesised on the solid phase (C).
Figure 3
Figure 3
Modified allylamine traps 2, 15–22.
Figure 4
Figure 4
Site‐selective installation of a range of Lys PTMs via visible‐light‐mediated, desulfurative C(sp3)–C(sp3) bond formation. [a] Photochemical set‐up 1 (blue LEDs). [b] Photochemical set‐up 2 (PhotoRedOx Box); isolated yields shown.
Figure 5
Figure 5
Site‐selective installation of an N ϵ Ac sidechain into recombinantly expressed Ub 35 (K48C mutant).
See this image and copyright information in PMC

References

    1. Aebersold R., Agar J. N., Amster I. J., Baker M. S., Bertozzi C. R., Boja E. S., Costello C. E., Cravatt B. F., Fenselau C., Garcia B. A., Nat. Chem. Biol. 2018, 14, 206–214. - PMC - PubMed
    1. Walsh C. T., Garneau-Tsodikova S., G. J. Gatto Jr , Angew. Chem. Int. Ed. 2005, 44, 7342–7372; - PubMed
    2. Angew. Chem. 2005, 117, 7508–7539.
    1. Luo M., Chem. Rev. 2018, 118, 6656–6705. - PMC - PubMed
    1. Ali I., Conrad R. J., Verdin E., Ott M., Chem. Rev. 2018, 118, 1216–1252. - PMC - PubMed
    1. Chen Y., Sprung R., Tang Y., Ball H., Sangras B., Kim S. C., Falck J. R., Peng J., Gu W., Zhao Y., Mol. Cell. Proteomics 2007, 6, 812–819. - PMC - PubMed

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