In-Cell Sensitivity-Enhanced NMR of Intact Viable Mammalian Cells

Rupam Ghosh, Yiling Xiao, Jaka Kragelj, Kendra K. Frederick

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

NMR has the resolution and specificity to determine atomic-level protein structures of isotopically labeled proteins in complex environments, and with the sensitivity gains conferred by dynamic nuclear polarization (DNP), NMR has the sensitivity to detect proteins at their endogenous concentrations. However, DNP sensitivity enhancements are critically dependent on experimental conditions and sample composition. While some of these conditions are theoretically compatible with cellular viability, the effects of others on cellular sample integrity are unknown. Uncertainty about the integrity of cellular samples limits the utility of experimental outputs of in-cell experiments. Using several measures, we establish conditions that support DNP enhancements that can enable detection of micromolar concentrations of proteins in experimentally tractable times that are compatible with cellular viability. Taken together, we establish DNP-assisted MAS NMR as a technique for structural investigations of biomolecules in intact viable cells that can be phenotyped both before and after NMR experiments.

Original languageEnglish (US)
Pages (from-to)18454-18466
Number of pages13
JournalJournal of the American Chemical Society
Volume143
Issue number44
DOIs
StatePublished - Nov 10 2021

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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