Selective Protein Degradation through Tetrazine Ligation of Genetically Incorporated Unnatural Amino Acids

• Published in: Chemistry, an Asian journal Vol. 19; no. 23; pp. e202400824 - n/a
• Main Authors: Chen, Jinghao, Dai, Gaocan, Duan, Shixiang, Huang, Yang, Wu, Yi‐Lin, Xie, Zhiyong, Tsai, Yu‐Hsuan
• Format: Journal Article
• Language: English
• Published: Germany 02.12.2024
• Subjects: Alkenes - chemistry; Alkynes - chemistry; Amino Acids - chemistry; Amino Acids - metabolism; Genetic code expansion; Humans; Hydrophobic tagging; Molecular Structure; Proteasome Endopeptidase Complex - chemistry; Proteasome Endopeptidase Complex - metabolism; Proteins - chemistry; Proteins - metabolism; Proteolysis; Targeted protein degradation; Tetrazine ligation; Ubiquitin-Conjugating Enzymes - chemistry; Ubiquitin-Conjugating Enzymes - genetics; Ubiquitin-Conjugating Enzymes - metabolism; Ubiquitination; Unnatural amino acid; Genetic code expansion; Hydrophobic tagging; Tetrazine ligation; Unnatural amino acid; Targeted protein degradation
• ISSN: 1861-4728; 1861-471X
• DOI: 10.1002/asia.202400824

Small molecule‐responsive tags for targeted protein degradation are valuable tools for fundamental research and drug target validation. Here, we show that genetically incorporated unnatural amino acids bearing a strained alkene or alkyne functionality can act as a minimalist tag for targeted protein degradation. Specifically, we observed the degradation of strained alkene‐ or alkyne‐containing kinases and E2 ubiquitin‐conjugating enzymes upon treatment with hydrophobic tetrazine conjugates. The extent of the induced protein degradation depends on the identity of the target protein, unnatural amino acid, and tetrazine conjugate, as well as the site of the unnatural amino acid in the target protein. Mechanistic studies revealed proteins undergo proteasomal degradation after tetrazine tethering, and the identity of tetrazine conjugates influences the dependence of ubiquitination on protein degradation. This work provides an alternative approach for targeted protein degradation and mechanistic insight, facilitating the future development of more effective targeted protein degradation strategies. In this study, we demonstrate that genetically incorporated unnatural amino acids containing strained alkene or alkyne groups can serve as minimalist tags for targeted protein degradation. Specifically, we observed the degradation of kinases and E2 ubiquitin‐conjugating enzymes with strained alkene or alkyne functionalities upon treatment with hydrophobic tetrazine conjugates.