Direct Observation of Membrane-Associated H-Ras in the Native Cellular Environment by In-Cell ¹⁹F-NMR Spectroscopy.

ABSTRACT: Ras acts as a molecular switch to control intracellular signaling on the plasma membrane (PM). Elucidating how Ras associates with PM in the native cellular environment is crucial for understanding its control mechanism. Here, we used in-cell nuclear magnetic resonance (NMR) spectroscopy combined with site-specific ¹⁹F-labeling to explore the membrane-associated states of H-Ras in living cells. The site-specific incorporation of p-trifluoromethoxyphenylalanine (OCF₃Phe) at three different sites of H-Ras, i.e., Tyr32 in switch I, Tyr96 interacting with switch II, and Tyr157 on helix α5, allowed the characterization of their conformational states depending on the nucleotide-bound states and an oncogenic mutational state. Exogenously delivered ¹⁹F-label

SUBMITTER: Ikari M 

PROVIDER: S-EPMC10302746 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature