PXD051289

PXD051289 is an original dataset announced via ProteomeXchange.

Title	GAPDH K251 Succinylation Induced by Tobacco Smoking Coordinates Glycolysis and Glutamine Metabolism to Promote NSCLC Progression
Description	NSCLC Tissue Mass Spectrometry Analysis: Mass spectrometry analysis of NSCLC tissues was carried out as previously described67. Briefly, lung cancer tissues were treated with RIPA lysate, protein was extracted, and the protein sample was run on a 10% SDS-PAGE gel. After the protein molecular weight distribution was identified by Coomassie bright blue staining, the gel was divided into five equal parts. The gel sample was evaporated with acetonitrile and reconstituted in 1% (v/v) formic acid. The protein peptide was then digested by trypsin, and the peptide sample was injected into the mass spectrometer (model LTQ, Thermo Fisher). The mass spectrometer was calibrated using standard compounds and operated in the data-dependent mode, in which the instrument cycled between full MS scans (m/z 300–2000), and the MS data were collected by targeting MS scans on the ten most abundant ions occurring in the MS scan. Finally, the mass spectrometry raw data of NSCLC tissues were used for further analysis of differential proteins and succinylation modification. Detection and Analysis of GAPDH Succinylation：The tryptic peptides were dissolved in 0.1% formic acid (solvent A) and directly loaded onto a homemade reverse-phase analytical column (15-cm length, 75-μm i.d.). The gradient was from 6% to 23% solvent B (0.1% formic acid in 98% acetonitrile) over 16 min, 23% to 35% over 8 min, and climbing to 80% over 3 min then holding at 80% for the last 3 min, all at a constant flow rate of 400 nl/min on an EASY-nLC 1000 UPLC system (Thermo Fisher). The peptides were subjected to NSI source followed by tandem mass spectrometry (MS/MS) in Q ExactiveTM Plus (Thermo Fisher) coupled online to the UPLC. The electrospray voltage applied was 2.0 kV. The m/z scan range was 350 to 1800 for full scans and intact peptides were detected in the Orbitrap at a resolution of 70,000. Peptides were then selected for MS/MS using an NCE setting of 28 and the fragments were detected in the Orbitrap at a resolution of 17,500. A data-dependent procedure that alternated between one MS scan followed by 20 MS/MS scans with 15.0 s dynamic exclusion. The automatic gain control (AGC) was set at 5E4.
HostingRepository	iProX
AnnounceDate	2024-04-09
AnnouncementXML	Submission_2025-04-26_21:46:26.932.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Kun Wang
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	LTQ Orbitrap; Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2024-04-09 01:05:31	ID requested
⏵ 1	2025-04-26 21:46:27	announced

Dataset with its publication pending

submitter keyword: Succinylation, Glycolysis and Glutamine Metabolism, NSCLC

Wan Huang
contact affiliation	Fourth Military Medical University
contact email	huangwan@fmmu.edu.cn
lab head
Kun Wang
contact affiliation	Fourth Military Medical University
contact email	827950581@qq.com
dataset submitter

iProX dataset URI