PXD038764

PXD038764 is an original dataset announced via ProteomeXchange.

Title	Comparative and temporal characterization of LPS and blue light induced TLR4 signal transduction and gene expression in optogenetically manipulated endothelial cells
Description	In endothelial cells (ECs), stimulation of Toll-like receptor 4 (TLR4) by the endotoxin lipopolysaccharide (LPS) induces the release of diverse pro-inflammatory mediators beneficial in controlling bacterial infections. However, their systemic secretion is a main driver of sepsis and chronic inflammatory diseases. Since distinct and rapid TLR4 signaling induction is difficult to achieve by LPS due to its poorly controllable binding and non-specific affinity to other surface molecules and receptors, we engineered new light-oxygen-voltage-sensing (LOV) domain-based optogenetic endothelial cell lines (opto-TLR4-LOV LEC and opto-TLR4-LOV HUVEC) that allow precise temporal and reversible activation of TLR4 signaling pathways. Using quantitative mass spectrometry, RT-qPCR and Western Blot analysis, we show that pro-inflammatory proteins were not only expressed differently, but also had a different time course when the cells were stimulated with light or LPS. Additional functional assays demonstrated that light induction promoted chemotaxis of THP-1 cells, disruption of the EC-monolayer and transmigration. In contrast, ECs incorporating a truncated version of the TLR4 extracellular domain (opto-TLR4 ΔECD2-LOV LEC) revealed high basal activity with fast exploitation of cell signaling system upon illumination. We conclude that the established optogenetic cell lines are well suited to induce rapid and precise photoactivation of TLR4, allowing receptor-specific studies.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:47:20.026.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD038764
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Benjamin Neuditschko
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2022-12-12 22:43:58	ID requested
1	2023-05-10 14:08:51	announced
⏵ 2	2023-11-14 08:47:25	announced	2023-11-14: Updated project metadata.

10.6019/PXD038764;

Stierschneider A, Neuditschko B, Colleselli K, Hundsberger H, Herzog F, Wiesner C, Comparative and Temporal Characterization of LPS and Blue-Light-Induced TLR4 Signal Transduction and Gene Expression in Optogenetically Manipulated Endothelial Cells. Cells, 12(5):(2023) [pubmed]

submitter keyword: lipopolysaccharide, chemotaxis,endothelial cells, mass spectrometry based proteomics, transmigration, Toll-like receptor 4, optogenetic control, pro-inflammatory proteins

Franz Herzog
contact affiliation	Institute Krems Bioanalytics, IMC University of Applied Sciences Krems, 3500 Krems, Austria
contact email	franz.herzog@fh-krems.ac.at
lab head
Benjamin Neuditschko
contact affiliation	Institute Krems Bioanalytics, IMC University of Applied Sciences Krems, 3500 Krems, Austria
contact email	benjamin.neuditschko@fh-krems.ac.at
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD038764
     1. Label: PRIDE project
     2. Name: Comparative and temporal characterization of LPS and blue light induced TLR4 signal transduction and gene expression in optogenetically manipulated endothelial cells