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Dynamic Nuclear Polarization with Electron Decoupling in Intact Human Cells and Cell Lysates

J Phys Chem B. 2020; 
Judge PT, Sesti EL, Price LE, Albert BJ, Alaniva N, Saliba EP, Halbritter T, Sigurdsson ST, Kyei GB, Barnes AB.
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Molecular Biology Reagents coli) cells electroporated with a PGEX- 4T plasmid (Genscript, Piscataway, NJ) bearing an insert coding for the C1b domain of the δ isoform of protein kinase C (PKC-δ) were incubated in Super Optimal Broth (S. Get A Quote

摘要

Dynamic nuclear polarization (DNP) is used to improve the inherently poor sensitivity of nuclear magnetic resonance spectroscopy by transferring spin polarization from electrons to nuclei. However, DNP radicals within the sample can have detrimental effects on nuclear spins close to the polarizing agent. Chirped microwave pulses and electron decoupling (eDEC) attenuate these effects in model systems, but this approach is yet to be applied to intact cells or cellular lysates. Herein, we demonstrate for the first time exceptionally fast 1H T1DNP times of just 200 and 300 ms at 90 and 6 K, respectively, using a newly synthesized methylated trityl radical within intact human cells. We further demonstrate that eDEC ... More

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