PXD045845-1

PXD045845 is an original dataset announced via ProteomeXchange.

Title	Structural basis for the modulation of MRP2 activity by phosphorylation and drugs
Description	Chemotherapy resistance by ABC transporters and secondary transporters is well established. Liver cancer is hard to treat by chemotherapy due to the presence of the multidrug resistance-associated protein 2 (MRP2/ABCC2) in addition to other transporters. The activity of MRP2 can be modulated by phosphorylation of its regulatory domain but also by drug-drug interactions. Here, we have determined the cryo-EM structure of MRP2 from Rattus norvegicus in an autoinhibited state (nucleotide-free) and in the presence of the drug modulator probenecid (uricosuric agent). The autoinhibited conformation is caused by the partially phosphorylated regulatory domain that folds within the transmembrane domain. Combining, in vitro phosphorylation of MRP2, mass spectrometry and kinetics, we show that full phosphorylation of the regulatory domain is required to relieve the autoinhibitory state due to steric clashes between the phosphate groups and the transmembrane domain. The drug bound state, has identified two drug binding sites that has allowed us to propose how probenecid modulates the activity of the transporter in addition to drug-drug interactions. Overall, we have structurally characterised the clinically relevant transporter MRP2, and we provide evidence on its modulation by intracellular kinases but also in the presence of external drugs. Finally, we mapped the mutations that cause Dubin–Johnson syndrome onto the structure and gain an understanding on their contribution in MRP2 dysfunction.
HostingRepository	PRIDE
AnnounceDate	2024-02-05
AnnouncementXML	Submission_2024-02-05_03:20:52.032.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Graeme Benstead-Hume
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-10-02 01:19:11	ID requested
⏵ 1	2024-02-05 03:20:52	announced
2	2024-10-22 06:28:34	announced	2024-10-22: Updated project metadata.

Dataset with its publication pending

submitter keyword: MRP2 activataion, phosphorylation

Jyoti Choudhary
contact affiliation	Functional Proteomics, ICR
contact email	jyoti.choudhary@icr.ac.uk
lab head
Graeme Benstead-Hume
contact affiliation	ICR
contact email	ghume@icr.ac.uk
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/2024/02/PXD045845

PRIDE project URI

• PRIDE
  1. PXD045845
     1. Label: PRIDE project
     2. Name: Structural basis for the modulation of MRP2 activity by phosphorylation and drugs