Investigation of Proteome-Tetrazine Reactivity for a Highly Selective Tetrazine Ligation in Live Cells
Tetrazine has been widely used in bio-orthogonal click chemistry for diverse biological applications due to its short reaction time and excellent bio-orthogonality. For efficient click reaction, the stability of tetrazine in physiological conditions is one of the key issues in biological applications. However, the reactions between tetrazine and biomolecules have barely been studied. Here, we investigated nonspecific proteome labeling by tetrazine derivatives. Systematic investigation of proteome reactivities of 23 tetrazine derivatives showed their structure-dependent proteome reactivities. We further investigated the relationship between the proteome reactivity of tetrazine derivatives and selectivity of <i>in situ</i> tetrazine-<i>trans</i>-cyclooctene (<b>Tz-TCO</b>) click chemistry-mediated fluorescent labeling of BTK protein in live cells. Intriguingly, a tetrazine derivative <b>SiR-Tz20</b> with minimal proteome reactivity showed a highly selective BTK labeling efficiency in live cells and in an <i>ex vivo</i> mouse model. Our results demonstrate that the proteome reactivity of tetrazine derivatives is critical for their selectivity in click reaction toward accurate fluorescent protein imaging in live cells.
Authors
- Juhee Hahm
- Harry C. Hwang
- Woon Ju Song (ORCID: https://orcid.org/0000-0003-0434-2684)
- Junhyeong Yim (ORCID: https://orcid.org/0000-0001-6709-0926)
- Jagadeesh Kumar Gangasani (ORCID: https://orcid.org/0000-0002-6386-4434)
- Soyeon Lee (ORCID: https://orcid.org/0000-0002-4562-0938)
- Hyunuk Eom (ORCID: https://orcid.org/0000-0002-2387-7046)
- Ju Young Park
- Hankum Park (ORCID: https://orcid.org/0000-0001-5795-3249)
- Jun‐Young Park (ORCID: https://orcid.org/0000-0002-2586-4732)
- Sanghee Lee (ORCID: https://orcid.org/0000-0002-7765-5916)
- Jongmin Park (ORCID: https://orcid.org/0000-0001-8338-9346)
- Eunha Kim (ORCID: https://orcid.org/0000-0002-0766-696X)
- Kyungmin Lee (ORCID: https://orcid.org/0000-0002-5425-4212)
- V. Arun
- Jeehee Lee (ORCID: https://orcid.org/0000-0002-1206-5114)
- Minju Lee (ORCID: https://orcid.org/0009-0002-0417-3489)
- Hana Cho (ORCID: https://orcid.org/0000-0002-3591-2013)
Institutions
- Seoul National University (KR)
- Kangwon National University (KR)
- Biocon (Switzerland) (CH)
- University of Dental Medicine (MM)
- Kyung Hee University (KR)
- Dimensional Imaging (United Kingdom) (GB)
- Advanced Institute of Convergence Technology (KR)
- Fusion (United States) (US)
- Institute for Molecular Science (JP)
- Materials Research Center (IN)
- Fusion Academy (US)
- Genomics (United Kingdom) (GB)
- PRG S&Tech (South Korea)
- Hanyang University (KR)
- Ajou University (KR)
- Korea Institute of Science and Technology (KR)
Publication Details
- Journal
- ACS Central Science
- Published
- 2025-05-06
- DOI
- https://doi.org/10.1021/acscentsci.5c00525
- Citations
- 7
- Primary Topic
- Click Chemistry and Applications
- Type
- article
- Field-Weighted Citation Impact
- 6.55
Funders
- Seoul National University
- Ministry of Education
- Korea Institute of Science and Technology
- National Research Foundation of Korea
- Gyeonggi-do Regional Research Center
- Ministry of Science and ICT, South Korea
- Samsung