PXD055459

PXD055459 is an original dataset announced via ProteomeXchange.

Title	Microproteomics of PCBP2 biomolecular condensates
Description	PCBP2 condensates were isolated by FAPS. To identify the PCBP2 condensates signature, we performed a mass spectrometry analysis and compared the pre-sorted and post-sorted fractions.For each sorting experiment, cells were cultured to approximately 80-90% confluence in three 15-cm dishes. Cells were pelleted in PBS by centrifugation at 1,000 × g for 3 min at RT and immediately flash-frozen in liquid nitrogen overnight. Unless otherwise stated, the following steps were conducted at 4°C or on ice. Cell pellets were resuspended in lysis buffer (50 mM Tris [pH 7.4], 1 mM EDTA, 150 mM NaCl, 0.2% Triton X-100), containing 65 U/mL RNase inhibitor and EDTA-free protease inhibitor cocktail. To facilitate lysis, the extracts were passed 20 times through a 25G syringe needle, with a total incubation time of 20 min. Lysates were spun at 200 x g for 5 min to remove nuclei. To digest remaining DNA contaminants, the supernatants were treated with RQ1 RNase-free DNase for 30 min at RT, and then centrifuged at 10,000 x g for 7 min. The resulting pellets were resuspended in 2 mL of lysis buffer, and the organelle-enriched fraction obtained at 10,000 x g was designated as the pre-sorted fraction. From this fraction, PCBP2-biomolecular condensates were sorted on a cell sorter (Bigfoot, Invitrogen) equipped with a 100 μm nozzle and operating at a pressure of 30 psi. Particles were identified based on their forward-scattered light (FSC) and mCherry fluorescence, utilizing a 561 nm excitation laser and a 615/24 nm bandpass filter. The sorting window for PCBP2-biomolecular condensates was specifically set to exclude mCherry-SH-SY5Y fluorescent particles while retaining mCherry-labeled PCBP2 condensates. The mCherry-PCBP2 fraction represented 8-10% of total events and was subsequently collected. These fractions were centrifuged at 10,000 × g for 7 min, and the pellets were then analyzed as the post-sorted fractions.
HostingRepository	iProX
AnnounceDate	2024-08-30
AnnouncementXML	Submission_2025-09-26_02:09:14.437.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Lu Wang
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2024-09-01 22:56:14	ID requested
⏵ 1	2025-09-26 02:09:14	announced

Dataset with its publication pending

submitter keyword: PCBP2 condensates, FAPS

Guojun Chen
contact affiliation	the First Affiliated Hospital of Chongqing Medical University
contact email	GJChen@hospital.cqmu.edu.cn
lab head
Lu Wang
contact affiliation	the First Affiliated Hospital of Chongqing Medical University
contact email	luwangcqmu@stu.cqmu.edu.cn
dataset submitter

iProX dataset URI