Abstract
Ever-increasing evidence confirms the role of O-GlcNAcase (OGA) in mediating cell growth and development as well as pathology and underscores the importance of developing sensitive and selective chemical tools for the study of OGA biology. Here, based on our previously developed protein-encapsulation strategy, we designed and synthesized a series of fluorogenic probes based on resorufin, and assembled their composites with human serum albumin (HSA) for the selective detection of endogenous OGA activity in live cells. We show that host–guest self-assembly with HSA significantly enhances the OGA sensitivity of the probes in aqueous solution and cells. The structure of the complex of a glycoprobe and HSA was resolved by small-angle X-ray scattering. We demonstrate that the replacement of the acetyl group in GlcNAc with a propionyl group results in selectivity for OGA over hexosaminidases (HEX) that unselectively hydrolyze hexosamines. This allows us to differentiate between two cell lines with different endogenous OGA and HEX expression levels, and selectively detect OGA activity in live cells.
| Original language | English |
|---|---|
| Journal | Chemical Science |
| Early online date | 17 Feb 2026 |
| DOIs | |
| Publication status | E-pub ahead of print - 17 Feb 2026 |
Data Availability Statement
All data supporting this research are available as part of the supplementary information (SI). Supplementary information: experimental methods, additional figures, and spectra of new compounds. See DOI: https://doi.org/10.1039/d5sc06843f.Funding
The authors thank the Natural National Science Foundation of China (No. 92253306, 82130099 and 22477030), Science and Technology Commission of Shanghai Municipality (grant No.24DX1400200), the International Cooperation Program of Shanghai Science and Technology (No.23490711600), the Fundamental Research Funds for the Central Universities (222201717003), the Programme of Introducing Talents of Discipline to Universities (B16017), the National Natural Science Foundation of Shanghai Science and Technology (No.24ZR1415400), the Shanghai Oriental Talents youth Program (No. QNKJ2024010),the Shanghai Xuhui District Hospital Local Cooperation Project (23XHYD-20), the Open Funding Project of the State Key Laboratory of Fine Chemicals, Dalian University of Technology (KF 2402), State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082, P. R. China, Ministry of Education Key Laboratory on Signaling Regulation and Targeting Therapy of Liver Cancer (Naval Medical University) (Grant. 2023-MEKLLC-MS/ZD-00*) and Shandong Laboratory Program (SYS202205) for financial support. T.D.J. wishes to thank the University of Bath and the Open Research Fund of the School of Chemistry and Chemical Engineering, Henan Normal University (2020ZD01) for support. The Research Center of Analysis and Test of East China University of Science and Technology was gratefully acknowledged for assistance in analytical experiments. The authors also thank the staff members of BL19U2 beamline (https://cstr.cn/31129.02.NFPS.BL19U2) at the National Facility for Protein Science in Shanghai (https://cstr.cn/31129.02.NFPS) for providing technical support and assistance in data collection and analysis.
ASJC Scopus subject areas
- General Chemistry
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