Proteomics

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Arabidopsis endomembrane enrichments: Affinity enrichment of mCherry bait

ABSTRACT: Our aim was to dissect subcellular trafficking routes by enriching for partially overlapping subpopulations of endosomal proteomes associated with endomembrane markers. We selected RABD2a/ARA5, RABF2b/ARA7, RABF1/ARA6, and RABG3f as markers for combinations of the Golgi, trans-Golgi Network (TGN), Early Endosomes (EE), secretory vesicles, Late Endosomes (LE), Multivesicular Bodies (MVB) compartments and tonoplast. As comparisons we used Golgi transport 1 (GOT1), which localises to the Golgi, Clathrin Light Chain 2 (CLC2) labelling Clathrin-coated vesicles and pits and the Vesicle Associated Membrane Protein 711 (VAMP711) present at the tonoplast. We developed an easy-to-use method by refining published protocols based on affinity purification of fluorescent fusion constructs to these seven subcellular marker proteins in Arabidopsis thaliana seedlings.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

SUBMITTER: Alex Jones  

LAB HEAD: Alexandra M. E. Jones

PROVIDER: PXD001884 | Pride | 2015-04-22

REPOSITORIES: Pride

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Dataset's files

Source:
Action DRS
OT_141126_12.raw Raw
OT_141126_12_mCherry (F004688).mzid_OT_141126_12_mCherry_(F004688).pride.mgf.gz Mzid
OT_141126_12_mCherry (F004688).pride.mztab.gz Mztab
OT_141126_12_mCherryF004688.mzid.gz Mzid
OT_141126_12_mCherryF004688.mzid_OT_141126_12_mCherry_F004688.MGF Mzid
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Publications

Identification of Regulatory and Cargo Proteins of Endosomal and Secretory Pathways in Arabidopsis thaliana by Proteomic Dissection.

Heard William W   Sklenář Jan J   Tomé Daniel F A DF   Robatzek Silke S   Jones Alexandra M E AM  

Molecular & cellular proteomics : MCP 20150421 7

The cell's endomembranes comprise an intricate, highly dynamic and well-organized system. In plants, the proteins that regulate function of the various endomembrane compartments and their cargo remain largely unknown. Our aim was to dissect subcellular trafficking routes by enriching for partially overlapping subpopulations of endosomal proteomes associated with endomembrane markers. We selected RABD2a/ARA5, RABF2b/ARA7, RABF1/ARA6, and RABG3f as markers for combinations of the Golgi, trans-Golg  ...[more]

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