PXD003162 is an
original dataset announced via ProteomeXchange.
Dataset Summary
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 |
Dataset History
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. |
Publication List
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] |
Keyword List
curator keyword: Biological |
submitter keyword: Chloroplast, Native Molecular Weight, Protein Complex, Oligomeric State, Native Gel |
Contact List
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 | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD003162
- Label: PRIDE project
- Name: Shotgun Proteomics of BN-PAGE-Resolved Chloroplasts