PXD051937

PXD051937 is an original dataset announced via ProteomeXchange.

Title	APEX2-based proximity labelling of Staphylococcus aureus receptors on human lung microvascular endothelial cells
Description	Staphylococcus aureus bacteria were decorated with modified ascorbate peroxidase APEX2-YFP-CWT via the cell wall targeting (CWT) domain of lysostaphin. Cytosol of transgenic HeLa cells expressing APEX2-YFP-CWT was generated by ultracentrifugation and living bacteria were incubated in the cytosol. The bacteria were stringently washed and added to either untreated human lung microvascular endothelial cells (HuLEC) or to cells pre-treated with bacterial sphingomyelinase (β-toxin), ionomycin, or amitriptyline. After 15 min, H2O2 was added for 1 min, then the biotinylation reaction was stopped by addition Stop Solution (containing sodium azide, sodium ascorbate, and Trolox in PBS). The cells then were scraped off the substratum, centrifuged, and lysed in RIPA buffer. To remove excess biotin phenol, lysates were transferred into a 3 kDa cutoff Amicon concentrator and 5 ml RIPA buffer was added. Solution was concentrated by centrifugation to 500-1000 µL. This step was repeated once. The lysates then were incubated overnight with magnetic streptavidin beads, followed by extensive washing and eventually biotinylated proteins were eluted by addition of Laemmli buffer containing an excess of biotin.
HostingRepository	PRIDE
AnnounceDate	2025-03-13
AnnouncementXML	Submission_2025-03-13_03:57:29.371.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Martin Fraunholz
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Staphylococcus aureus; NCBI TaxID: 1280;
ModificationList	biotinylated residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2024-05-02 02:54:23	ID requested
⏵ 1	2025-03-13 03:57:29	announced

10.1128/MBIO.03654-24;

submitter keyword: human lung microvascular endothelial cells, receptor identification, APEX2 proximity labeling,Staphylococcus aureus

Martin J Fraunholz
contact affiliation	Chair of Microbiology Biocenter, University of Würzburg D-97074 Würzburg, GERMANZ
contact email	martin.fraunholz@uni-wuerzburg.de
lab head
Martin Fraunholz
contact affiliation	Chair of Microbiology, Biocenter, University Würzburg
contact email	martin.fraunholz@uni-wuerzburg.de
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/2025/03/PXD051937

PRIDE project URI

• PRIDE
  1. PXD051937
     1. Label: PRIDE project
     2. Name: APEX2-based proximity labelling of Staphylococcus aureus receptors on human lung microvascular endothelial cells