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

[Image: see text] Limited chemical shift dispersion represents a significant barrier to studying multistate equilibria of large membrane proteins by (19)F NMR. We describe a novel monofluoroethyl (19)F probe that dramatically increases the chemical shift dispersion. The improved conformational sensi...

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Autores principales: Huang, Yun, Reddy, Krishna D., Bracken, Clay, Qiu, Biao, Zhan, Wenhu, Eliezer, David, Boudker, Olga
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2023
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10119980/
https://www.ncbi.nlm.nih.gov/pubmed/37023263
http://dx.doi.org/10.1021/jacs.3c01003
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author Huang, Yun
Reddy, Krishna D.
Bracken, Clay
Qiu, Biao
Zhan, Wenhu
Eliezer, David
Boudker, Olga
author_facet Huang, Yun
Reddy, Krishna D.
Bracken, Clay
Qiu, Biao
Zhan, Wenhu
Eliezer, David
Boudker, Olga
author_sort Huang, Yun
collection PubMed
description [Image: see text] Limited chemical shift dispersion represents a significant barrier to studying multistate equilibria of large membrane proteins by (19)F NMR. We describe a novel monofluoroethyl (19)F 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) (19)F 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, (19)F NMR can guide sample preparation to discover and visualize novel conformational states and facilitate image analysis and three-dimensional (3D) classification.
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spelling pubmed-101199802023-04-22 Environmentally Ultrasensitive Fluorine Probe to Resolve Protein Conformational Ensembles by (19)F NMR and Cryo-EM Huang, Yun Reddy, Krishna D. Bracken, Clay Qiu, Biao Zhan, Wenhu Eliezer, David Boudker, Olga J Am Chem Soc [Image: see text] Limited chemical shift dispersion represents a significant barrier to studying multistate equilibria of large membrane proteins by (19)F NMR. We describe a novel monofluoroethyl (19)F 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) (19)F 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, (19)F NMR can guide sample preparation to discover and visualize novel conformational states and facilitate image analysis and three-dimensional (3D) classification. American Chemical Society 2023-04-06 /pmc/articles/PMC10119980/ /pubmed/37023263 http://dx.doi.org/10.1021/jacs.3c01003 Text en © 2023 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Huang, Yun
Reddy, Krishna D.
Bracken, Clay
Qiu, Biao
Zhan, Wenhu
Eliezer, David
Boudker, Olga
Environmentally Ultrasensitive Fluorine Probe to Resolve Protein Conformational Ensembles by (19)F NMR and Cryo-EM
title Environmentally Ultrasensitive Fluorine Probe to Resolve Protein Conformational Ensembles by (19)F NMR and Cryo-EM
title_full Environmentally Ultrasensitive Fluorine Probe to Resolve Protein Conformational Ensembles by (19)F NMR and Cryo-EM
title_fullStr Environmentally Ultrasensitive Fluorine Probe to Resolve Protein Conformational Ensembles by (19)F NMR and Cryo-EM
title_full_unstemmed Environmentally Ultrasensitive Fluorine Probe to Resolve Protein Conformational Ensembles by (19)F NMR and Cryo-EM
title_short Environmentally Ultrasensitive Fluorine Probe to Resolve Protein Conformational Ensembles by (19)F NMR and Cryo-EM
title_sort environmentally ultrasensitive fluorine probe to resolve protein conformational ensembles by (19)f nmr and cryo-em
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10119980/
https://www.ncbi.nlm.nih.gov/pubmed/37023263
http://dx.doi.org/10.1021/jacs.3c01003
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