PXD061317-1

PXD061317 is an original dataset announced via ProteomeXchange.

Title	Unveiling Cellular Communications through Rapid Pan-Membrane-Protein Labeling
Description	The dynamic protein distribution within and across the plasma membrane is pivotal in regulating cell communications. Yet, methods facilitating rapid labeling, applicable across diverse cell types, and amenable to multiplexed live imaging are scarce. We discovered that live mammalian cell surface proteins can be labeled using N-hydroxysuccinimide (NHS)-ester-based amine crosslinking of fluorescent dyes. Using mouse DC2.4 dendritic cells, human Jurkat T cells, and mouse primary lymphoma T cells as model systems, we revealed the membrane topology and membrane-associated cell communications previously challenging to capture by fluorescence microscopy techniques. In DC2.4 cells, we observed transient membrane protein accumulation at cell-cell contacts and their bidirectional migration patterns guided by membrane fibers. We further visualized the involvement of membrane fibers in the transfer of membrane-derived particles and ovalbumin, as well as caveolin-dependent endocytosis and subsequent transport of membrane labels in late endosomes and lysosomes. In addition, we demonstrate caveolin-1 phosphorylation-dependent insulin receptor endocytosis in HEK 293T cells. Volumetric structured illumination microscopy revealed changes in the membrane topology of Jurkat T cells in response to the activating surface, generation of membrane-derived microvesicles, and phagocytosis of CD3/CD28 Dynabeads on the glass surface. Finally, we demonstrate that the membrane label remains stable in the in vivo environment and evidence of intercellular membrane protein transfer from splenocytes from the donor T cell lymphoma SNF5 mouse to the healthy splenocytes in the C57/BL6 mouse 24 and 48-h after cell transfer. The ultrahigh membrane labeling density, coupled with high multiplexity and rapid labeling, enables innovative fluorescence microscopy applications to dissect membrane and membrane-derived structures in intercellular and intracellular communication both in vitro and in vivo.
HostingRepository	PRIDE
AnnounceDate	2025-02-28
AnnouncementXML	Submission_2025-02-28_10:34:41.066.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Daniel Abegg
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	timsTOF HT

Revision	Datetime	Status	ChangeLog Entry
0	2025-02-28 10:14:18	ID requested
⏵ 1	2025-02-28 10:34:41	announced

Dataset with its publication pending

submitter keyword: mouse, NHS-biotin, plasma membrane

Alexander Adibekian
contact affiliation	Department of Chemistry, University of Illinois Chicago, Chicago, USA
contact email	aadibeki@uic.edu
lab head
Daniel Abegg
contact affiliation	University of Illinois at Chicago
contact email	dabegg@uic.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/02/PXD061317

PRIDE project URI

• PRIDE
  1. PXD061317
     1. Label: PRIDE project
     2. Name: Unveiling Cellular Communications through Rapid Pan-Membrane-Protein Labeling