Issue 36, 2023

A high-field cellular DNP-supported solid-state NMR approach to study proteins with sub-cellular specificity

Abstract

Studying the structural aspects of proteins within sub-cellular compartments is of growing interest. Dynamic nuclear polarization supported solid-state NMR (DNP-ssNMR) is uniquely suited to provide such information, but critically lacks the desired sensitivity and resolution. Here we utilize SNAPol-1, a novel biradical, to conduct DNP-ssNMR at high-magnetic fields (800 MHz/527 GHz) inside HeLa cells and isolated cell nuclei electroporated with [13C,15N] labeled ubiquitin. We report that SNAPol-1 passively diffuses and homogenously distributes within whole cells and cell nuclei providing ubiquitin spectra of high sensitivity and remarkably improved spectral resolution. For cell nuclei, physical enrichment facilitates a further 4-fold decrease in measurement time and provides an exclusive structural view of the nuclear ubiquitin pool. Taken together, these advancements enable atomic interrogation of protein conformational plasticity at atomic resolution and with sub-cellular specificity.

Graphical abstract: A high-field cellular DNP-supported solid-state NMR approach to study proteins with sub-cellular specificity

Supplementary files

Article information

Article type
Edge Article
Submitted
25 apr 2023
Accepted
24 aug 2023
First published
05 sep 2023
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2023,14, 9892-9899

A high-field cellular DNP-supported solid-state NMR approach to study proteins with sub-cellular specificity

D. Beriashvili, R. Yao, F. D'Amico, M. Krafčíková, A. Gurinov, A. Safeer, X. Cai, M. P. C. Mulder, Y. Liu, G. E. Folkers and M. Baldus, Chem. Sci., 2023, 14, 9892 DOI: 10.1039/D3SC02117C

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