Thiocholine-Mediated One-Pot Peptide Ligation and Desulfurization

Sae Suzuki; Yuya Nakajima; Naoki Kamo; Akihisa Osakabe; Akimitsu Okamoto; Gosuke Hayashi; Hiroshi Murakami

doi:10.3390/molecules28093655

Thiocholine-Mediated One-Pot Peptide Ligation and Desulfurization

Molecules. 2023 Apr 22;28(9):3655. doi: 10.3390/molecules28093655.

Authors

Sae Suzuki¹, Yuya Nakajima¹, Naoki Kamo², Akihisa Osakabe³, Akimitsu Okamoto², Gosuke Hayashi¹, Hiroshi Murakami^{1

4}

Affiliations

¹ Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
² Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
³ Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
⁴ Institute of Nano-Life-Systems, Institutes of Innovation for Future Society, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.

Abstract

Thiol catalysts are essential in native chemical ligation (NCL) to increase the reaction efficiency. In this paper, we report the use of thiocholine in chemical protein synthesis, including NCL-based peptide ligation and metal-free desulfurization. Evaluation of thiocholine peptide thioester in terms of NCL and hydrolysis kinetics revealed its practical utility, which was comparable to that of other alkyl thioesters. Importantly, thiocholine showed better reactivity as a thiol additive in desulfurization, which is often used in chemical protein synthesis to convert Cys residues to more abundant Ala residues. Finally, we achieved chemical synthesis of two differently methylated histone H3 proteins via one-pot NCL and desulfurization with thiocholine.

Keywords: chemical protein synthesis; desulfurization; native chemical ligation; one-pot synthesis.

MeSH terms

Histones
Ligation
Peptides* / chemistry
Sulfhydryl Compounds / chemistry
Thiocholine*

Substances

Thiocholine
Peptides
Sulfhydryl Compounds
Histones

Grants and funding

This research was funded by the Japan Society for the Promotion of Science (JSPS), including KAKENHI JP21K20628, JP22H05172, and JP22H05178 (A. Osakabe); 19K22245 (A. Okamoto); 19H05287, 20H04704, 21H00278, and 22KK0128 (G.H.); JST PRESTO JPMJPR20K3 (A. Osakabe); and JPMJPR19K6 (G.H.). This work was also supported by AMED under grant numbers JP22ama121009 (G.H.) and JP21zf0127004 (H.M.) and by a donation from H.M.