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 pr

SUBMITTER: Beriashvili D 

PROVIDER: S-EPMC10510770 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature