High-pH structure of EmrE reveals the mechanism of proton-coupled substrate transport. Shcherbakov, A. A., Spreacker, P. J., Dregni, A. J., Henzler-Wildman, K. A., & Hong, M. Nature Communications, 13(1):991, December, 2022. ![High-pH structure of EmrE reveals the mechanism of proton-coupled substrate transport [link]](https://bibbase.org/img/filetypes/link.svg "link")
Paper doi abstract bibtex Abstract The homo-dimeric bacterial membrane protein EmrE effluxes polyaromatic cationic substrates in a proton-coupled manner to cause multidrug resistance. We recently determined the structure of substrate-bound EmrE in phospholipid bilayers by measuring hundreds of protein-ligand H N –F distances for a fluorinated substrate, 4-fluoro-tetraphenylphosphonium (F 4 -TPP + ), using solid-state NMR. This structure was solved at low pH where one of the two proton-binding Glu14 residues is protonated. Here, to understand how substrate transport depends on pH, we determine the structure of the EmrE-TPP complex at high pH, where both Glu14 residues are deprotonated. The high-pH complex exhibits an elongated and hydrated binding pocket in which the substrate is similarly exposed to the two sides of the membrane. In contrast, the low-pH complex asymmetrically exposes the substrate to one side of the membrane. These pH-dependent EmrE conformations provide detailed insights into the alternating-access model, and suggest that the high-pH conformation may facilitate proton binding in the presence of the substrate, thus accelerating the conformational change of EmrE to export the substrate.
@article{shcherbakov_high-ph_2022,
title = {High-{pH} structure of {EmrE} reveals the mechanism of proton-coupled substrate transport},
volume = {13},
issn = {2041-1723},
url = {https://www.nature.com/articles/s41467-022-28556-6},
doi = {10.1038/s41467-022-28556-6},
abstract = {Abstract
The homo-dimeric bacterial membrane protein EmrE effluxes polyaromatic cationic substrates in a proton-coupled manner to cause multidrug resistance. We recently determined the structure of substrate-bound EmrE in phospholipid bilayers by measuring hundreds of protein-ligand H
N
–F distances for a fluorinated substrate, 4-fluoro-tetraphenylphosphonium (F
4
-TPP
+
), using solid-state NMR. This structure was solved at low pH where one of the two proton-binding Glu14 residues is protonated. Here, to understand how substrate transport depends on pH, we determine the structure of the EmrE-TPP complex at high pH, where both Glu14 residues are deprotonated. The high-pH complex exhibits an elongated and hydrated binding pocket in which the substrate is similarly exposed to the two sides of the membrane. In contrast, the low-pH complex asymmetrically exposes the substrate to one side of the membrane. These pH-dependent EmrE conformations provide detailed insights into the alternating-access model, and suggest that the high-pH conformation may facilitate proton binding in the presence of the substrate, thus accelerating the conformational change of EmrE to export the substrate.},
language = {en},
number = {1},
urldate = {2022-10-11},
journal = {Nature Communications},
author = {Shcherbakov, Alexander A. and Spreacker, Peyton J. and Dregni, Aurelio J. and Henzler-Wildman, Katherine A. and Hong, Mei},
month = dec,
year = {2022},
keywords = {Proton-coupled substrate transport},
pages = {991},
}
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This structure was solved at low pH where one of the two proton-binding Glu14 residues is protonated. Here, to understand how substrate transport depends on pH, we determine the structure of the EmrE-TPP complex at high pH, where both Glu14 residues are deprotonated. The high-pH complex exhibits an elongated and hydrated binding pocket in which the substrate is similarly exposed to the two sides of the membrane. In contrast, the low-pH complex asymmetrically exposes the substrate to one side of the membrane. These pH-dependent EmrE conformations provide detailed insights into the alternating-access model, and suggest that the high-pH conformation may facilitate proton binding in the presence of the substrate, thus accelerating the conformational change of EmrE to export the substrate.","language":"en","number":"1","urldate":"2022-10-11","journal":"Nature Communications","author":[{"propositions":[],"lastnames":["Shcherbakov"],"firstnames":["Alexander","A."],"suffixes":[]},{"propositions":[],"lastnames":["Spreacker"],"firstnames":["Peyton","J."],"suffixes":[]},{"propositions":[],"lastnames":["Dregni"],"firstnames":["Aurelio","J."],"suffixes":[]},{"propositions":[],"lastnames":["Henzler-Wildman"],"firstnames":["Katherine","A."],"suffixes":[]},{"propositions":[],"lastnames":["Hong"],"firstnames":["Mei"],"suffixes":[]}],"month":"December","year":"2022","keywords":"Proton-coupled substrate transport","pages":"991","bibtex":"@article{shcherbakov_high-ph_2022,\n\ttitle = {High-{pH} structure of {EmrE} reveals the mechanism of proton-coupled substrate transport},\n\tvolume = {13},\n\tissn = {2041-1723},\n\turl = {https://www.nature.com/articles/s41467-022-28556-6},\n\tdoi = {10.1038/s41467-022-28556-6},\n\tabstract = {Abstract \n \n The homo-dimeric bacterial membrane protein EmrE effluxes polyaromatic cationic substrates in a proton-coupled manner to cause multidrug resistance. 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