PXD022185

PXD022185 is an original dataset announced via ProteomeXchange.

Title	Outward-facing structures of a multidrug ABC exporter with and without bound drug
Description	Multidrug ATP-Binding Cassette (ABC) exporters use an alternating access mechanism to translocate drugs across membranes, but the molecular mechanism of substrate release is still elusive due to the poor affinity of the drug-binding pocket for its ligand at this stage. Here, we bring new information on this mechanism by resolving two ATP-bound outward-facing conformations of the homodimer BmrA from Bacillus subtilis by X-ray crystallography with no drug bound and by cryo-EM in the presence of rhodamine 6G. This revealed the first structure of a multidrug pump with a substrate bound to the drug-binding pocket in a pre-release state. Two molecules of the dye are located in an hydrophobic region at the level of the outer leaflet between transmembrane (TM) helices 1, 2 and 6. Markedly, the X-ray structures display a half-closed TM1-2 region that is widely open in the cryo-EM one. Hydrogen-Deuterium exchange coupled to mass spectrometry (HDX-MS) and molecular dynamics simulations confirmed the high flexibility of this region, presumably driving the release of structurally divergent compounds.
HostingRepository	PRIDE
AnnounceDate	2021-03-25
AnnouncementXML	Submission_2021-03-25_07:25:09.293.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Julien Marcoux
SpeciesList	scientific name: Bacillus subtilis; NCBI TaxID: 1423;
ModificationList	monohydroxylated residue
Instrument	Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2020-10-26 05:12:45	ID requested
⏵ 1	2021-03-25 07:25:09	announced

Dataset with its publication pending

submitter keyword: membrane exporter crystal and cryo-EM structure, substrate release, transport mechanism, hydrogen-deuterium exchange, nanodiscs

Julien Marcoux
contact affiliation	Proteomics and Mass Spectrometry of Biomolecules, Institut de Pharmacologie et de Biologie Structurale (IPBS), Toulouse, France
contact email	julien.marcoux@ipbs.fr
lab head
Julien Marcoux
contact affiliation	IPBS
contact email	julien.marcoux@ipbs.fr
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/03/PXD022185

PRIDE project URI

• PRIDE
  1. PXD022185
     1. Label: PRIDE project
     2. Name: Outward-facing structures of a multidrug ABC exporter with and without bound drug