Dataset Information

Mass spectrometry analyzed human IL-33 interaction networks under various stimulations

ABSTRACT: C-terminal HA tagged hIL-33 were ectopically expressed in HEK293T cells for 24 hours, and demonstrated with indicating stimulations (Ctrl: without stimulations, Pap: 10ug/mL papain exposure for 30min, AS: 0.5mM arsenite exposure for 1hour). Cell lysates were collected for HA-immunoprecipitation and further mass spectrometry analyze.

ORGANISM(S): Homo Sapiens

SUBMITTER: Bing Sun

PROVIDER: PXD033460 | iProX | Tue Apr 26 00:00:00 BST 2022

REPOSITORIES: iProX

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Publications

Allergen protease-activated stress granule assembly and gasdermin D fragmentation control interleukin-33 secretion.

Chen Wen W Chen Shuangfeng S Yan Chenghua C Zhang Yaguang Y Zhang Ronghua R Chen Min M Zhong Shufen S Fan Weiguo W Zhu Songling S Zhang Danyan D Lu Xiao X Zhang Jia J Huang Yuying Y Zhu Lin L Li Xuezhen X Lv Dawei D Fu Yadong Y Iv Houkun H Ling Zhiyang Z Ma Liyan L Jiang Hai H Long Gang G Zhu Jinfang J Wu Dong D Wu Bin B Sun Bing B

Nature immunology 20220706 7

Interleukin-33 (IL-33), an epithelial cell-derived cytokine that responds rapidly to environmental insult, has a critical role in initiating airway inflammatory diseases. However, the molecular mechanism underlying IL-33 secretion following allergen exposure is not clear. Here, we found that two cell events were fundamental for IL-33 secretion after exposure to allergens. First, stress granule assembly activated by allergens licensed the nuclear-cytoplasmic transport of IL-33, but not the secret ...[more]

PMID: 35794369

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Project description:IL-33 is a nuclear cytokine from the IL-1 family that plays important roles in health and disease. Under healthy conditions, IL-33 is constitutively expressed to high levels in the nucleus of producing cells in various human and mouse tissues. The extracellular function of IL-33 cytokine has been well documented, but it remains unclear whether intracellular nuclear IL-33 has additional functions in the nucleus. Here, we used a global proteomic approach based on quantification of 5000 individual proteins by high-resolution mass spectrometry to compare the extracellular and intracellular roles of IL-33 in primary human endothelial cells, a major source of IL-33 protein in human tissues. Large-scale analysis of protein expression was performed either after stimulation of the cells with the IL-33 mature form IL-3395-270 (during 6h or 24h) or after siRNA knockdown of intracellular IL-33 (two experiments, each with a different pool of distinct siRNAs, noted siRNA1 and siRNA2). In each case, proteins were fractionated by 1D SDS-PAGE in 12 gel bands, and label-free quantitative analysis was performed. The present dataset contains the files for the two experiments of knockdown of endogenous nuclear IL-33 expression: - RNA silencing strategy 1. Knockdown of endogenous nuclear IL-33 expression was performed with a pool of four distinct siRNAs (Dharmacon ON-TARGETplus SMARTpool IL-33 siRNAs) that have been specifically modified for efficient silencing of the target gene with reduced off-target effects. Cells transfected with these siRNA duplexes (si1) were compared with those transfected with the provided controls (CTsi1). Three independent biological replicates (noted _A, _B, _C) were prepared and analyzed for each condition, leading to 6 different samples. Each of them was fractionated into 12 gel bands analyzed by nanoLC-MS/MS, leading to 72 raw files. - RNA silencing strategy 2. The second knockdown strategy was based on the use of an independent pool of three siRNAs targeting IL-33, predesigned by another provider using new and critical siRNA design rules (Sigma MISSION Predesigned Il-33 siRNAs based on Rosetta siRNA design algorithm). Cells transfected with these siRNA duplexes (si2) were compared with those transfected with the provided controls (CTsi2). Three independent biological replicates (noted _A, _B, _C) were prepared and analyzed for each condition, leading to 6 different samples. Each of them was fractionated into 12 gel bands analyzed by nanoLC-MS/MS, leading to 72 raw files.

Dataset Information

Mass spectrometry analyzed human IL-33 interaction networks under various stimulations

Publications

Allergen protease-activated stress granule assembly and gasdermin D fragmentation control interleukin-33 secretion.

Similar Datasets

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