PXD003162

PXD003162 is an original dataset announced via ProteomeXchange.

Title	Shotgun Proteomics of BN-PAGE-Resolved Chloroplasts
Description	Blue Native-PAGE resolves protein complexes in their native state. When coupled with immunoblotting, it can be used to identify the presence of high molecular weight complexes at high resolution for any protein, given a suitable antibody. To identify proteins in high molecular weight complexes on a large-scale and to bypass the requirement for specific antibodies, we applied a tandem mass spectrometry (MS/MS) approach to BN-PAGE-resolved chloroplasts. Fractionation of wild-type chloroplasts into 6 bands allowed identification and label-free quantification of 1000 chloroplast proteins with native molecular weight resolution. Significantly, our approach achieves a depth of identification comparable to traditional shotgun proteomic analyses of chloroplasts, indicating much of the known chloroplast proteome is amenable to our ‘mass western’ approach to BN-PAGE. The coupling of BN-PAGE to MS/MS allows for a large-scale comparison of protein complexes between samples and the identification of proteins in high molecular weight complexes. In parallel we have analyzed chloroplasts from a leaf reticulate mutant (re-6) to demonstrate the possibility for comparative analyses. Our results provide a useful resource for the chloroplast community and this strategy is anticipated to be widely adaptable to other sub-cellular compartments.
HostingRepository	PRIDE
AnnounceDate	2017-10-24
AnnouncementXML	Submission_2017-10-24_03:41:23.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Peter Lundquist
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	No PTMs are included in the dataset
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2015-11-09 01:56:15	ID requested
1	2016-12-23 08:56:17	announced
⏵ 2	2017-10-24 03:41:24	announced	Updated project metadata.

Lundquist PK, Mantegazza O, Stefanski A, St, ü, hler K, Weber APM, Surveying the Oligomeric State of Arabidopsis thaliana Chloroplasts. Mol Plant, 10(1):197-211(2017) [pubmed]

curator keyword: Biological

submitter keyword: Chloroplast, Native Molecular Weight, Protein Complex, Oligomeric State, Native Gel

Andreas P. M Weber
contact affiliation	Heinrich Heine University - Institute for Plant Biochemistry
contact email	andreas.weber@hhu.de
lab head
Peter Lundquist
contact affiliation	Samuel Roberts Noble Foundation
contact email	PKL5@cornell.edu
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/2016/12/PXD003162

PRIDE project URI

• PRIDE
  1. PXD003162
     1. Label: PRIDE project
     2. Name: Shotgun Proteomics of BN-PAGE-Resolved Chloroplasts