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. 2023 Mar 16;29(16):e202202503.
doi: 10.1002/chem.202202503. Epub 2023 Feb 14.

Cysteine-Selective Modification of Peptides and Proteins via Desulfurative C-C Bond Formation

Rhys C Griffiths  1 , Frances R Smith  1 , Diyuan Li  1 , Jasmine Wyatt  2 , David M Rogers  1 , Jed E Long  3 , Lola M L Cusin  4 , Patrick J Tighe  4 , Robert Layfield  3 , Jonathan D Hirst  1 , Manuel M Müller  2 , Nicholas J Mitchell  1
Affiliations

Cysteine-Selective Modification of Peptides and Proteins via Desulfurative C-C Bond Formation

Rhys C Griffiths et al. Chemistry. .

Abstract

The site-selective modification of peptides and proteins facilitates the preparation of targeted therapeutic agents and tools to interrogate biochemical pathways. Among the numerous bioconjugation techniques developed to install groups of interest, those that generate C(sp3 )-C(sp3 ) bonds are significantly underrepresented despite affording proteolytically stable, biogenic linkages. Herein, a visible-light-mediated reaction is described that enables the site-selective modification of peptides and proteins via desulfurative C(sp3 )-C(sp3 ) bond formation. The reaction is rapid and high yielding in peptide systems, with comparable translation to proteins. Using this chemistry, a range of moieties is installed into model systems and an effective PTM-mimic is successfully integrated into a recombinantly expressed histone.

Keywords: bioconjugation; cysteine; desulfurization; post-translational modifications; site-selective.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Cys‐selective peptide/protein modification via C(sp3)−C(sp3) bond formation.
Figure 2
Figure 2
A. Cys‐selective installation of PTM mimics/desired groups via C(sp3)−C(sp3) bond formation. [a] Isolated yields shown using standard blue LEDs. [b] Trap 17 only partially soluble in reaction solvent. B. Crude HPLC trace for the reaction of 8 with 2 after 5 mins under protocol B conditions run in the PhotoRedOx Box. C. Time course of reaction progress using the PhotoRedOx Box. [c] Entry 2 conditions used for model peptide 8. D. [d] Reactions run in the PhotoRedOx Box. [e] Isolated yield in brackets.
Figure 3
Figure 3
Installation of a sulfonate group and elimination to afford an isoprenyl handle followed by site‐selective fluorination.
Figure 4
Figure 4
Modification of ubiquitin (Ub) K48C (32) with trimethylammonium (11) and biotin (12) traps.
Figure 5
Figure 5
A. Installation of a trimethyl Lys sidechain into histone H4K20C (35). B. Deconvoluted ESI MS of crude material shows the majority desired product (36; calculated mass [M+H]+ 11,305.4) and the minority presence of the undesired by‐product carrying Ala at the target site (37; calculated mass [M+H]+ 11,178.3). C. Western blot using anti‐histone H4 (trimethyl K20) antibody to demonstrate effective recognition of mimics 36 and 38. D. Desulfurized protein 37 and mimic 38.
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