PXD066215

PXD066215 is an original dataset announced via ProteomeXchange.

Title	Proteomic profiling of GFP-tagged PIN-LIKES (PILS2, PILS3, PILS6) in Arabidopsis root extracts reveals ERAD components as major interactors
Description	This dataset describes a proteomic analysis of proteins co-immunoprecipitated with functional GFP-tagged PILS2, PILS3, and PILS6 fusion proteins in Arabidopsis thaliana roots. PILS (PIN-LIKES) proteins are auxin transport facilitators that localize to the endoplasmic reticulum (ER). To identify proteins that associate with PILS2, PILS3, and PILS6 under physiological conditions, we performed GFP-based immunoprecipitation (IP) followed by mass spectrometry (MS). Protein extracts were prepared from root tissues of Arabidopsis Col-0 (wild-type) and PILS2-, PILS3-, and PILS6-overexpressing transgenic lines expressing N- or C-terminal GFP fusion proteins. Plant material was ground in liquid nitrogen, and total proteins were extracted using aTris Buffer containing 1% CHAPS, 20 mM Tris-HCl (pH 7.5), 10 mM MgCl₂, 15 mM EGTA, 75 mM NaCl, 1 mM DTT, 0.1% Tween-20, and protease inhibitors. Extracts were centrifuged, and the supernatants were quantified using the Bradford assay. 2mg of protein were subjected to immunoprecipitation using anti-GFP magnetic beads, Col-0 (non-tagged) extracts were used as negative controls. Co-immunoprecipitated proteins were then processed and analyzed by mass spectrometry.
HostingRepository	PRIDE
AnnounceDate	2025-08-21
AnnouncementXML	Submission_2025-08-21_05:49:45.671.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Markus Hartl
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	phosphorylated residue
Instrument	Q Exactive HF; Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2025-07-16 07:47:01	ID requested
⏵ 1	2025-08-21 05:49:46	announced

Dataset with its publication pending

submitter keyword: Arabidopsis thaliana, Roots, ERAD

Jürgen Kleine-Vehn
contact affiliation	Faculty of Biology,Molecular Plant Physiology, Universit of Freiburg,Schänzlestr. 1, 79104 Freiburg¸Germany
contact email	juergen.kleine-vehn@biologie.uni-freiburg.de
lab head
Markus Hartl
contact affiliation	Max Perutz Labs, Mass Spectrometry Facility, Vienna Biocenter Campus (VBC), Dr. Bohr-Gasse 9, Vienna, Austria
contact email	proteomics@maxperutzlabs.ac.at
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD066215
     1. Label: PRIDE project
     2. Name: Proteomic profiling of GFP-tagged PIN-LIKES (PILS2, PILS3, PILS6) in Arabidopsis root extracts reveals ERAD components as major interactors