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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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2018
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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. |
format | Online Article Text |
id | pubmed-5852267 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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