期刊
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
卷 143, 期 44, 页码 18454-18466出版社
AMER CHEMICAL SOC
DOI: 10.1021/jacs.1c06680
关键词
-
资金
- National Institutes of Health [NS-111236]
- Welch Foundation [I1923_20170325]
- Lupe Murchison Foundation
- Kinship Foundation (Searle Scholars Program)
- NCI Cancer Center Support Grant [1P30 CA142543-01]
NMR combined with DNP enhances sensitivity and resolution for detecting proteins, but sensitivity gains are influenced by experimental conditions and sample composition, leading to uncertainty about cellular sample integrity.
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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据