PXD018866-1

PXD018866 is an original dataset announced via ProteomeXchange.

Title	A peptidisc-based AP/MS approach to explore the interactome of the bacterial Sec translocon
Description	We recently presented the peptidisc membrane mimetic for the global reconstitution of membrane proteomes into water-soluble detergent-free particles, termed peptidisc library. We here develop a method that combines the peptidisc library with affinity pulldown and mass spectrometry (AP/MS) to explore the membrane interactome at native expression level, which minimizes artifacts due to protein overproduction or exposure to detergents. Using the Sec translocon as a case study, we identify an expanded holo-translocon complex, termed the HMD complex, which consists of 9 different membrane subunits - SecYEG, SecDFyajC, YidC, plus YfgM and PpiD. This super-complex can be biochemically isolated in peptidisc but not in detergent. We also detect associations of the HMD complex with the outer membrane Bam complex and we discover a novel unannotated inner membrane protein named YibN. This interactome analysis at native expression level points out the membrane complexes that are dissociated in detergent, yet stable enough to be captured in peptidiscs. We believe this information is critical for the design of biochemical procedures that will enable their successful purification.
HostingRepository	PRIDE
AnnounceDate	2023-03-22
AnnouncementXML	Submission_2023-03-22_09:30:34.456.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	SadhnaPhanse
SpeciesList	scientific name: Escherichia coli (strain K12 / W3110 / ATCC 27325 / DSM 5911); NCBI TaxID: 316407;
ModificationList	iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2020-04-28 09:50:46	ID requested
⏵ 1	2023-03-22 09:30:34	announced

Dataset with its publication pending

submitter keyword: Mass Spectrometry, Affinity tagging, Sec translocon, Protein Translocation,Peptidisc, Protein complex analysis, Membrane mimetics, Membrane protein complexes , Protein-Protein Interactions, Affinity Purification/Mass Spectrometry (AP/MS)

FranckDuong
contact affiliation	Department of Biochemistry and Molecular Biology, Faculty of Medicine, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada
contact email	fduong@mail.ubc.ca
lab head
SadhnaPhanse
contact affiliation	University of Toronto; University of Regina
contact email	srphanse@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD018866
     1. Label: PRIDE project
     2. Name: A peptidisc-based AP/MS approach to explore the interactome of the bacterial Sec translocon