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  Identification of covalent modifications regulating immune signaling complex composition and phenotype

Frauenstein, A., Ebner, S., Hansen, F. M., Sinha, A., Phulphagar, K., Swatek, K., et al. (2021). Identification of covalent modifications regulating immune signaling complex composition and phenotype. Molecular Systems Biology, 17(7): e10125. doi:10.15252/msb.202010125.

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 Creators:
Frauenstein, Annika1, Author              
Ebner, Stefan1, Author              
Hansen, Fynn M.2, Author              
Sinha, Ankit2, Author              
Phulphagar, Kshiti1, Author              
Swatek, Kirby3, Author              
Hornburg, Daniel4, Author
Mann, Matthias2, Author              
Meissner, Felix1, Author              
Affiliations:
1Meissner, Felix / Experimental Systems Immunology, Max Planck Institute of Biochemistry, Max Planck Society, ou_2149678              
2Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565159              
3Schulman, Brenda / Molecular Machines and Signaling, Max Planck Institute of Biochemistry, Max Planck Society, ou_2466699              
4external, ou_persistent22              

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Free keywords: PROTEIN-INTERACTION NETWORKS; Q EXACTIVE HF; NF-KAPPA-B; MASS-SPECTROMETRY; CELL-DEATH; KINASE; PURIFICATION; INFLAMMASOME; TNF; SYSTEMBiochemistry & Molecular Biology; mass spectrometry; posttranslational modifications; protein interactions; proteomics; signaling networks;
 Abstract: Cells signal through rearrangements of protein communities governed by covalent modifications and reversible interactions of distinct sets of proteins. A method that identifies those post-transcriptional modifications regulating signaling complex composition and functional phenotypes in one experimental setup would facilitate an efficient identification of novel molecular signaling checkpoints. Here, we devised modifications, interactions and phenotypes by affinity purification mass spectrometry (MIP-APMS), comprising the streamlined cloning and transduction of tagged proteins into functionalized reporter cells as well as affinity chromatography, followed by MS-based quantification. We report the time-resolved interplay of more than 50 previously undescribed modification and hundreds of protein-protein interactions of 19 immune protein complexes in monocytes. Validation of interdependencies between covalent, reversible, and functional protein complex regulations by knockout or site-specific mutation revealed ISGylation and phosphorylation of TRAF2 as well as ARHGEF18 interaction in Toll-like receptor 2 signaling. Moreover, we identify distinct mechanisms of action for small molecule inhibitors of p38 (MAPK14). Our method provides a fast and cost-effective pipeline for the molecular interrogation of protein communities in diverse biological systems and primary cells.

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Language(s): eng - English
 Dates: 2021
 Publication Status: Published online
 Pages: 21
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000678530300001
DOI: 10.15252/msb.202010125
 Degree: -

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Title: Molecular Systems Biology
Source Genre: Journal
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Publ. Info: London : Nature Pub. Group
Pages: - Volume / Issue: 17 (7) Sequence Number: e10125 Start / End Page: - Identifier: ISSN: 1744-4292
CoNE: https://pure.mpg.de/cone/journals/resource/1000000000021290