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Influence of quaternary cation compound on the size of the Escherichia coli small multidrug resistance protein, EmrE

EmrE is a member of the small multidrug resistance (SMR) protein family in Escherichia coli. It confers resistance to a wide variety of quaternary cation compounds (QCCs) as an efflux transporter driven by the transmembrane proton motive force. We have expressed hexahistidinyl (His(6)) – myc epitope...

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Autores principales: Qazi, S. Junaid S., Turner, Raymond J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5852267/
https://www.ncbi.nlm.nih.gov/pubmed/29552647
http://dx.doi.org/10.1016/j.bbrep.2018.02.001
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author Qazi, S. Junaid S.
Turner, Raymond J.
author_facet Qazi, S. Junaid S.
Turner, Raymond J.
author_sort Qazi, S. Junaid S.
collection PubMed
description EmrE is a member of the small multidrug resistance (SMR) protein family in Escherichia coli. It confers resistance to a wide variety of quaternary cation compounds (QCCs) as an efflux transporter driven by the transmembrane proton motive force. We have expressed hexahistidinyl (His(6)) – myc epitope tagged EmrE, extracted it from membrane preparations using the detergent n-dodecyl-β-D-maltopyranoside (DDM), and purified it using nickel-affinity chromatography. The size of the EmrE protein, in DDM environment, was then examined in the presence and absence of a range of structurally different QCC ligands that varied in their chemical structure, charge and shape. We used dynamic light scattering and showed that the size and oligomeric state distributions are dependent on the type of QCC. We also followed changes in the Trp fluorescence and determined apparent dissociation constants (K(d)). Overall, our in vitro analyses of epitope tagged EmrE demonstrated subtle but significant differences in the size distributions with different QCC ligands bound.
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spelling pubmed-58522672018-03-16 Influence of quaternary cation compound on the size of the Escherichia coli small multidrug resistance protein, EmrE Qazi, S. Junaid S. Turner, Raymond J. Biochem Biophys Rep Research Article EmrE is a member of the small multidrug resistance (SMR) protein family in Escherichia coli. It confers resistance to a wide variety of quaternary cation compounds (QCCs) as an efflux transporter driven by the transmembrane proton motive force. We have expressed hexahistidinyl (His(6)) – myc epitope tagged EmrE, extracted it from membrane preparations using the detergent n-dodecyl-β-D-maltopyranoside (DDM), and purified it using nickel-affinity chromatography. The size of the EmrE protein, in DDM environment, was then examined in the presence and absence of a range of structurally different QCC ligands that varied in their chemical structure, charge and shape. We used dynamic light scattering and showed that the size and oligomeric state distributions are dependent on the type of QCC. We also followed changes in the Trp fluorescence and determined apparent dissociation constants (K(d)). Overall, our in vitro analyses of epitope tagged EmrE demonstrated subtle but significant differences in the size distributions with different QCC ligands bound. Elsevier 2018-02-20 /pmc/articles/PMC5852267/ /pubmed/29552647 http://dx.doi.org/10.1016/j.bbrep.2018.02.001 Text en © 2018 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Qazi, S. Junaid S.
Turner, Raymond J.
Influence of quaternary cation compound on the size of the Escherichia coli small multidrug resistance protein, EmrE
title Influence of quaternary cation compound on the size of the Escherichia coli small multidrug resistance protein, EmrE
title_full Influence of quaternary cation compound on the size of the Escherichia coli small multidrug resistance protein, EmrE
title_fullStr Influence of quaternary cation compound on the size of the Escherichia coli small multidrug resistance protein, EmrE
title_full_unstemmed Influence of quaternary cation compound on the size of the Escherichia coli small multidrug resistance protein, EmrE
title_short Influence of quaternary cation compound on the size of the Escherichia coli small multidrug resistance protein, EmrE
title_sort influence of quaternary cation compound on the size of the escherichia coli small multidrug resistance protein, emre
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5852267/
https://www.ncbi.nlm.nih.gov/pubmed/29552647
http://dx.doi.org/10.1016/j.bbrep.2018.02.001
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