- Secondary multidrug (Mdr) transporters utilize ion concentration gradients to actively remove antibiotics and other toxic compounds from cells. The model Mdr transporter MdfA from Escherichia coli exchanges dissimilar drugs for protons. The transporter should open at the cytoplasmic side to enable access of drugs into the Mdr recognition pocket. Here we show that the cytoplasmic rim around the Mdr recognition pocket represents a previously overlooked important regulatory determinant in MdfA. We demonstrate that increasing the positive charge of the electrically asymmetric rim dramatically inhibits MdfA activity and sometimes even leads to influx of planar, positively charged compounds, resulting in drug sensitivity. Our results suggest that unlike the mutants with the electrically modified rim, the membrane-embedded wild-type MdfA exhibits a significant probability of an inward-closed conformation, which is further increased by drug binding. Since MdfA binds drugs from its inward-facing environment, these results are intriguing and raise the possibility that the transporter has a sensitive, drug-induced conformational switch, which favors an inward-closed state.
MetadatenAuthor: | Elia Zomot, Eliane Hadas Yardeni, Attilio Vittorio Vargiu, Heng Keat Tam, Giuliano MallociORCiD, Venkata Krishnan Ramaswamy, Michal Perach, Paolo Ruggerone, Klaas Martinus PosORCiD, Eitan Bibi |
---|
URN: | urn:nbn:de:hebis:30:3-502611 |
---|
DOI: | https://doi.org/10.1016/j.jmb.2018.02.026 |
---|
ISSN: | 1089-8638 |
---|
ISSN: | 0022-2836 |
---|
Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/29530612 |
---|
Parent Title (English): | Journal of molecular biology |
---|
Publisher: | Elsevier |
---|
Place of publication: | Amsterdam [u. a.] |
---|
Contributor(s): | Nieng Yan |
---|
Document Type: | Article |
---|
Language: | English |
---|
Year of Completion: | 2018 |
---|
Date of first Publication: | 2018/03/09 |
---|
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
---|
Release Date: | 2019/05/16 |
---|
Tag: | MdfA; major facilitator superfamily; multidrug resistance; multidrug transport; secondary transport |
---|
Volume: | 430 |
---|
Issue: | 9 |
---|
Page Number: | 18 |
---|
First Page: | 1368 |
---|
Last Page: | 1385 |
---|
Note: | © 2018 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
---|
HeBIS-PPN: | 451291247 |
---|
Institutes: | Biochemie, Chemie und Pharmazie / Biochemie und Chemie |
---|
Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften |
---|
Sammlungen: | Universitätspublikationen |
---|
Licence (German): | Creative Commons - Namensnennung-Nicht kommerziell - Keine Bearbeitung 4.0 |
---|