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Environmentally Ultrasensitive Fluorine Probe to Resolve Protein Conformational Ensembles by ¹⁹F NMR and Cryo-EM.

ABSTRACT: Limited chemical shift dispersion represents a significant barrier to studying multistate equilibria of large membrane proteins by ¹⁹F NMR. We describe a novel monofluoroethyl ¹⁹F probe that dramatically increases the chemical shift dispersion. The improved conformational sensitivity and line shape enable the detection of previously unresolved states in one-dimensional (1D) ¹⁹F NMR spectra of a 134 kDa membrane transporter. Changes in the populations of these states in response to ligand binding, mutations, and temperature correlate with population changes of distinct conformations in structural ensembles determined by single-particle cryo-electron microscopy (cryo-EM). Thus, ¹⁹F NMR can guide sample preparation to discover and visualize novel conformational states and facilitate image analysis and three-dimensional (3D) classification.

SUBMITTER: Huang Y

PROVIDER: S-EPMC10119980 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature

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Environmentally Ultrasensitive Fluorine Probe to Resolve Protein Conformational Ensembles by 19F NMR and Cryo-EM.

Huang Yun Y Reddy Krishna D KD Bracken Clay C Qiu Biao B Zhan Wenhu W Eliezer David D Boudker Olga O

Journal of the American Chemical Society 20230406 15

Limited chemical shift dispersion represents a significant barrier to studying multistate equilibria of large membrane proteins by 19F NMR. We describe a novel monofluoroethyl 19F probe that dramatically increases the chemical shift dispersion. The improved conformational sensitivity and line shape enable the detection of previously unresolved states in one-dimensional (1D) 19F NMR spectra of a 134 kDa membrane transporter. Changes in the populations of these sta ...[more]

PMID: 37023263

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