期刊
CHEMICAL SCIENCE
卷 13, 期 47, 页码 14157-14164出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/d2sc04668g
关键词
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资金
- National Natural Science Foundation of China [22174099, 21871210, 21572161, 31500684]
- Dutch Research Council (NWO) [700.26.121, 700.10.443, 718.015.001, ECHO.018.067]
- uNMR-NL
- National Roadmap Large-Scale NMR Facility of the Netherlands [184.032.207]
- Science & Technology Projects of Tianjin [20JCZDJC00050]
- Hefei National Laboratory for Physical Sciences at the Microscale [KF2020001]
Cellular dynamic nuclear polarization is an effective method for overcoming the sensitivity limitations of solid-state nuclear magnetic resonance spectroscopy, allowing atomic-level characterization of biomolecules in their native environments. Researchers have synthesized a highly bioresistant and hydrophilic polarizing agent, StaPol-1, which shows excellent performance in reductive environments and is suitable for DNP ssNMR investigations at high magnetic fields.
Cellular dynamic nuclear polarization (DNP) has been an effective means of overcoming the intrinsic sensitivity limitations of solid-state nuclear magnetic resonance (ssNMR) spectroscopy, thus enabling atomic-level biomolecular characterization in native environments. Achieving DNP signal enhancement relies on doping biological preparations with biradical polarizing agents (PAs). Unfortunately, PA performance within cells is often limited by their sensitivity to the reductive nature of the cellular lumen. Herein, we report the synthesis and characterization of a highly bioresistant and hydrophilic PA (StaPol-1) comprising the trityl radical OX063 ligated to a gem-diethyl pyrroline nitroxide via a rigid piperazine linker. EPR experiments in the presence of reducing agents such as ascorbate and in HeLa cell lysates demonstrate the reduction resistance of StaPol-1. High DNP enhancements seen in small molecules, proteins and cell lysates at 18.8 T confirm that StaPol-1 is an excellent PA for DNP ssNMR investigations of biomolecular systems at high magnetic fields in reductive environments.
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