Multi-Dimensional Structure and Dynamics Landscape of Proteins in Mammalian Cells Revealed by In-cell NMR

Governing function, half life and subcellular localization, the 3D structure and dynamics of proteins are in nature constantly changing in a tightly regulated manner to fulfill the physiological and adaptive requirements of the cells. To find evidence for this hypothesis, we applied in-cell NMR to three folded model proteins and propose that the splitting of cross peaks are due to distinct structural states that arise from multiple target binding co-existing inside mammalian cells as a result of subcellular localisation including distinct cell compartments. In addition to peak splitting, we observed NMR signal intensity attenuations indicative of transient interactions with other molecules and dynamics on the microsecond to millisecond time scale.