PXD014995

PXD014995 is an original dataset announced via ProteomeXchange.

Title	The Function of Ectopic ATP Synthase in Extracellular Vesicles.
Description	Ectopic adenosine triphosphate synthase (eATP synthase) was found on the cell surface to generate extracellular ATP in various cancer cells. Recently using proteomics approach, several reports identified ATP synthase in extracellular vesicles (EVs) which are important in cell-cell communications. However, the functions of ATP synthase in EVs are still unclear. Here we increased the expression of eATP synthase on the plasma membrane of lung cancer A549 cells via 0.1% FBS starvation and collected EVs including microvesicles and exosomes from the culture medium using serial centrifugation. Both of them were assuredly isolated and confirmed by transmission electron microscopy, nano tracking analysis, and specific EV markers. We also used western blot to demonstrate the presence of ATP synthase in EVs. Besides, dot blot experiment revealed that the F1 portion faced towards extracellular space using anti-ATP beta subunit antibody. Under starvation, the expression of eATP synthase was enhanced on cell surface and exosomes as well as increased EV secretion. To further investigate the functions, we first elucidated whether eATP synthase on EV membrane produced ATP by ATP luciferase analysis. Our data showed that there was no significant ATP production. Moreover, we performed proteomic analysis using dimethyl labeling and LC-MS/MS to compare the difference between EVs released by A549 control and starvation. We identified a total of 1504 and 869 proteins in microvesicles and exosomes, respectively. Using 1.5- and 0.67-fold change as the thresholds, there were 171 and 46 differential expressed proteins in microvesicles and exosomes, respectively. Seven ATP synthase subunits in microvesicles and five in exosomes were identified. The proteomic data were further analyzed using gene set enrichment analysis (GSEA) within the C2 (curated gene sets) subclasses and setting 0.25 as FDR cutoff to find out biological functions related to eATP synthase. Among these up-regulated terms, metabolism, cell cycle and apoptosis were enriched, which indicating that starvation influences cell behavior. Additionally, the immune response is upregulated strikingly in exosomes within C2 gene set. One of these immune response pathways, MHC-1 antigen presentation, was previously proved to interact with eATP synthase which could be recognized by several immune cells.
HostingRepository	PRIDE
AnnounceDate	2023-05-22
AnnouncementXML	Submission_2023-05-21_23:38:28.338.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	ChangNai-Wen
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2019-08-12 05:31:23	ID requested
⏵ 1	2023-05-21 23:38:28	announced

Dataset with its publication pending

submitter keyword: Ectopic ATP synthases

extracellular vesicles

proteomic analysis

serum depletion

human lymphoma Jurkat T cells

Hsueh-FenJuan
contact affiliation	Department of Life Science,National Taiwan University, Taipei, Taiwan
contact email	yukijuan@gmail.com
lab head
ChangNai-Wen
contact affiliation	Institude of Molecular and Cellular Biology, National Taiwan University
contact email	r06b43004@ntu.edu.tw
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD014995
     1. Label: PRIDE project
     2. Name: The Function of Ectopic ATP Synthase in Extracellular Vesicles.