Functional Raman Probes for Detecting Enzyme Activities Based on Aggregation Control
Visualizing specific enzyme activities in living systems is important in biomedical research, and Raman imaging probes are particularly suitable for simultaneous multiplexed detection of plural enzyme activities due to their narrow signal peak widths. Here, we present a molecular design strategy for enzyme-activity-detecting Raman probes based on control of aggregation. The probe itself is soluble and diffusive in aqueous solution due to its hydrophilic substrate moiety, but hydrolysis by the target enzyme affords a hydrophobic product that forms aggregates, thereby increasing the local dye concentration, which in turn results in a stronger Raman signal that enables visualization of the enzyme activity. We validated the molecular design by developing Raman probes targeting aminopeptidase, glycosidase and carboxypeptidase. The vibrational frequency can be shifted by isotope-editing, and the developed probes were successfully applied to visualize the activities of aminopeptidase and glycosidase in live cultured cells and spheroids.
Authors
- Hiroyoshi Fujioka (ORCID: https://orcid.org/0000-0003-1904-0566)
- Yasuyuki Ozeki (ORCID: https://orcid.org/0000-0002-9004-0799)
- Yasuteru Urano (ORCID: https://orcid.org/0000-0002-1220-6327)
- Spencer J. Spratt (ORCID: https://orcid.org/0000-0002-5429-8856)
- Minoru Kawatani (ORCID: https://orcid.org/0000-0002-0192-4366)
- Aoi Ishikawa
- Hiroki Ito (ORCID: https://orcid.org/0000-0003-4745-5750)
- Ryosuke Kojima (ORCID: https://orcid.org/0000-0002-3792-9222)
- Yoshihiro Misawa
- Mako Kamiya (ORCID: https://orcid.org/0000-0002-5592-1849)
- Momoko Okinaka
- Reika Otake
Institutions
- Tokyo Institute of Technology (JP)
- Shanghai Institute for Science of Science (CN)
- Integrated Oncology (United States) (US)
- Life Science Institute (JP)
- Center for High Pressure Science and Technology Advanced Research (CN)
- Institute of Science Tokyo (JP)
- The University of Tokyo (JP)
- Graduate School USA (US)
- Kumamoto University (JP)
Publication Details
- Journal
- Analytical Chemistry
- Published
- 2025-08-27
- DOI
- https://doi.org/10.1021/acs.analchem.5c02231
- Citations
- 1
- Primary Topic
- Spectroscopy Techniques in Biomedical and Chemical Research
- Type
- article
- Field-Weighted Citation Impact
- 1.63
Funders
- Japan Agency for Medical Research and Development
- Ministry of Education, Culture, Sports, Science and Technology
- Secom Science and Technology Foundation
- University of Tokyo
- Astellas Pharma
- Mochida Memorial Foundation for Medical and Pharmaceutical Research
- Daiichi Sankyo Foundation of Life Science
- Astellas Foundation for Research on Metabolic Disorders
- Nakatani Foundation for Advancement of Measuring Technologies in Biomedical Engineering
- Research Foundation for Opto-Science and Technology
- Japan Society for the Promotion of Science
- Core Research for Evolutional Science and Technology
- JST-Mirai Program
- ACT-X
- Moonshot Research and Development Program
- Fusion Oriented REsearch for disruptive Science and Technology
- Daiichi Sankyo Europe