Dataset Information

Pericyte-specific secretome profiling in hypoxia using TurboID in an in vitro spheroid-model

ABSTRACT: This project utilizes TurboID to tag secreted proteins with biotin for subsequent cell specific analysis of the secretome from co-culture conditions

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Brain, Brain Pericyte

DISEASE(S): Disease Free

SUBMITTER: Charlotte Welinder

LAB HEAD: Andreas Enström

PROVIDER: PXD044689 | Pride | 2024-05-07

REPOSITORIES: Pride

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Publications

Pericyte-Specific Secretome Profiling in Hypoxia Using TurboID in a Multicellular In Vitro Spheroid-Model.

Enström Andreas A Carlsson Robert R Buizza Carolina C Lewi Marvel M Paul Gesine G

Molecular & cellular proteomics : MCP 20240503

Cellular communication within the brain is imperative for maintaining homeostasis and mounting effective responses to pathological triggers like hypoxia. However, a comprehensive understanding of the precise composition and dynamic release of secreted molecules has remained elusive, confined primarily to investigations using isolated monocultures. To overcome these limitations, we utilized the potential of TurboID, a non-toxic biotin ligation enzyme, to capture and enrich secreted proteins speci ...[more]

PMID: 38705386

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Project description:In the intracellular pathogen Brucella spp., the activation of the stringent response, a global regulatory network providing rapid adaptation to a variety of growth-affecting stress conditions such as nutrient deficiency, is essential for replication in the host. A single, bi-functional enzyme Rsh catalyzes synthesis and hydrolysis of the alarmone (p)ppGpp, responsible for differential gene expression under stringent conditions. cDNA microarray analysis allowed characterization of the transcriptional profiles of the B. suis 1330 wild-type and rsh mutant in a synthetic minimal medium, partially mimicking the nutrient-poor environment of the intramacrophagic vacuole. A total of 379 genes (11.6% of the genome) were differentially expressed in a rsh-dependent manner, of which 52% were up-regulated and 48% were down-regulated. The pleiotropic character of the response was confirmed, as the genes encoded factors belonging to various functional groups, comprising an important number of transcriptional regulators, cell envelope proteins, stress factors, transport systems, and energy metabolism proteins. Several virulence genes were under the positive control of (p)ppGpp. Methionine was the only amino acid whose biosynthesis was absolutely dependent on stringent response in B. suis. The study illustrated the complexity of the processes involved in adaptation to nutrient starvation, and contributed to a better understanding of the correlation between stringent response and Brucella virulence. Most interestingly, it clearly indicated (p)ppGpp-dependent cross-talk between at least three stress responses playing a central role in Brucella adaptation to the host: nutrient, oxidative, and low-oxygen stress. Two-condition experiment, wild-type against rsh mutant. Biological replicates: 4 wild-type, 4 rsh mutant, independently grown in minimal medium (under nutrient starvation condition) and harvested. One replicate per array. Please note that Channel 1 (Cy3) on each hybridization was a universal reference (Uref) used as a reference point to merge two (2 channel arrays) into a virtual 2 channel array with wild-type versus rsh mutant.

Dataset Information

Pericyte-specific secretome profiling in hypoxia using TurboID in an in vitro spheroid-model

Publications

Pericyte-Specific Secretome Profiling in Hypoxia Using TurboID in a Multicellular In Vitro Spheroid-Model.

Similar Datasets

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