In the context of studying the role of the COP9 signalosome (CSN) in neuroinflammation and ischemic neuronal damage, we studied the effect of the cullin NEDDylation state-modifying drugs MLN4924 and CSN5i-3 in BV2 microglial cells, an immortalized murine cell line featuring many of the characteristics of primary microglia. Owing to its potent inhibitory effect on the NEDDylation cascade, MLN4924 exhibits a CSN5-like anti-inflammatory activity. Csn5i-3 is a small molecule inhibitor that specifically binds to CSN5, while it resides in the CSN holocomplex and blocks its deNEDDylase activity, thus leading to an accumulation of NEDDylated cullins. We performed untargeted mass spectrometry-based proteomics of the cell lysates after treating BV2 cells with these drugs under basal culture stress conditions. The proteomic analysis revealed that MLN4924 and CSN5i-3 substantially altered the microglial proteome.
Export PNG
Views: 52
Created: 9th Jul 2024 at 13:04
This item has not yet been tagged.
None
Version History
Version 1 (earliest) Created 9th Jul 2024 at 13:04 by Rainer Malik
No revision comments
Related items
Projects: SyNergy - published datasets
Institutions: DZNE
Projects: SyNergy - published datasets
Institutions: Klinikum der Universität München
Projects: SyNergy - published datasets
Institutions: Klinikum der Universität München
Public web page: Not specified
Organisms: Mus musculus, Rattus norvegicus, Homo sapiens, Macaca mulatta, Sus scrofa
Submitter: Rainer Malik
Studies: ACSL3/GABARAPL2 interactor, ADAM10-Mediated Ectodomain Shedding Is an Essential Driver of Podocyte D..., AMPK/Lysosomal Damage/Galectins, An optimized quantitative proteomics method establishes the cell type-re..., Autophagosome content profiling, Autophagy cargo/CD4+ T cell proliferation, BAG3-mediated autophagy, Basic Fibroblast Growth Factor 2-Induced Proteome Changes Endorse Lewy B..., Beneficial Effect of ACI-24 Vaccination on Aβ Plaque Pathology and Micro..., C9orf72/UBR5/Quality control, CCNF/ALS, CNN2/Lysosome damage, COP9 signalosome/Ischemic stroke, Cell-type-specific profiling of brain mitochondria reveals functional an..., Deciphering sources of PET signals in the tumor microenvironment of glio..., Endothelial Aging, Experimental evidence for temporal uncoupling of brain Aβ deposition and..., Fibrillar Aβ triggers microglial proteome alterations and dysfunction in..., Loss of NPC1 enhances phagocytic uptake and impairs lipid trafficking in..., Lysosomal targeting of TAPL, NDP52 interactome, Neuronal differentiation of LUHMES cells is accompanied by substantial c..., NrCAM is a marker for substrate-selective activation of ADAM10 in Alzhei..., Pro-inflammatory activation following demyelination is required for myel..., Protein content of autophagic vesicles, Proteolysis enhanced proximity proteomics, Proteomic and lipidomic profiling of demyelinating lesions identifies fa..., Proteomic profiling in cerebral amyloid angiopathy reveals an overlap wi..., Proteomics of inflammasome activation for rapid immune responses, Proteomics of mouse brain endothelium uncovers dysregulation of vesicula..., Reactivated endogenous retroviruses promote protein aggregate spreading, Signal peptide peptidase-like 2c impairs vesicular transport and cleaves..., Signatures of glial activity can be detected in the CSF proteome, Spatial proteomics in three-dimensional intact specimens, Spatial proteomics reveals secretory pathway disturbances caused by neur..., TBK1/ALS, TECPR2/secretory pathway, Targeting the TCA cycle can ameliorate widespread axonal energy deficien..., The Alzheimer's disease-linked protease BACE1 modulates neuronal IL-6 si..., The Alzheimer's disease-linked protease BACE2 cleaves VEGFR3 and modulat..., The intramembrane protease SPPL2c promotes male germ cell development by..., The pseudoprotease iRhom1 controls ectodomain shedding of membrane prote..., The tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor com..., UBE2QL1/Lysosome damage, UBQLN2/ALS
Assays: Affinity purification coupled with mass spectrometry proteomics, Affinity purification coupled with mass spectrometry proteomics, Affinity purification coupled with mass spectrometry proteomics, Affinity purification coupled with mass spectrometry proteomics, Affinity purification coupled with mass spectrometry proteomics, Affinity purification coupled with mass spectrometry proteomics, Affinity purification coupled with mass spectrometry proteomics, Bottom-up proteomics, Gel-based experiment, Proteomics analysis of brain endothelial cells, Proteomics of Inflammasome, SWATH MS, Shotgun Proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics, Shotgun proteomics
Snapshots: Snapshot 1
In the context of studying the role of the COP9 signalosome (CSN) in neuroinflammation and ischemic neuronal damage, we studied the effect of the cullin NEDDylation state-modifying drugs MLN4924 and CSN5i-3 in BV2 microglial cells, an immortalized murine cell line featuring many of the characteristics of primary microglia. Owing to its potent inhibitory effect on the NEDDylation cascade, MLN4924 exhibits a CSN5-like anti-inflammatory activity. Csn5i-3 is a small molecule inhibitor that specifically ...
Snapshots: No snapshots
BV2 cells were cultured in full medium in 10 cm cell culture dishes until they were confluent. MLN4924 (500 nM), CSN5i-3 (1 μM), or solvent control (0.01% DMSO) were added and the culturing continued for 6 h. After the incubation, cells were washed once with PBS, removed from the cell culture plate with a scraper, collected in tubes, and centrifuged in 1.5 ml Eppendorf tubes for 3 min to remove remaining PBS buffer, snap-frozen, and stored at -80 °C until further processing. Thereafter cells were ...
Submitter: Rainer Malik
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Proteomics
Organisms: Mus musculus
SOPs: No SOPs
Data files: The COP9 signalosome reduces neuroinflammation ...
Snapshots: No snapshots
Abstract (Expand)
Authors: Y. Tian, J. Milic, L. S. Monasor, R. Chakraborty, S. Wang, Y. Yuan, Y. Asare, C. Behrends, S. Tahirovic, J. Bernhagen
Date Published: 19th Aug 2023
Publication Type: Journal
PubMed ID: 37597109
Citation: Cell Mol Life Sci. 2023 Aug 19;80(9):262. doi: 10.1007/s00018-023-04911-8.