Proteome and Phosphoproteome Analysis in TNF Long Term-Exposed Primary Human Monocytes
AbstractTo better understand the inflammation-associated mechanisms modulating and terminating tumor necrosis factor (TNF-)induced signal transduction and the development of TNF tolerance, we analyzed both the proteome and the phosphoproteome in TNF long term-incubated (i.e., 48 h) primary human monocytes using liquid chromatography-mass spectrometry. Our analyses revealed the presence of a defined set of proteins characterized by reproducible changes in expression and phosphorylation patterns in long term TNF-treated samples. In total, 148 proteins and 569 phosphopeptides were significantly regulated (103 proteins increased, 45 proteins decreased; 377 peptides with increased and 192 peptides with decreased phosphorylation). A variety of these proteins are associated with the non-canonical nuclear factor κB (NF-κB) pathway (nuclear factor κB (NFKB) 2, v-rel reticuloendotheliosis viral oncogene homolog (REL) B, indolamin-2,3-dioxygenase (IDO), kynureninase (KYNU)) or involved in the negative regulation of the canonical NF-κB system. Within the phosphopeptides, binding motifs for specific kinases were identified. Glycogen synthase kinase (GSK) 3 proved to be a promising candidate, since it targets NF-κB inhibiting factors, such as CCAAT/enhancer binding protein (C/EBP) β. Our experiments demonstrate that both proteome and phosphoproteome analysis can be effectively applied to study protein/phosphorylation patterns of primary monocytes. These results provide new regulatory candidates and evidence for a complex network of specific but synergistically acting/cooperating mechanisms enabling the affected cells to resist sustained TNF exposure and resulting in the resolution of inflammation. View Full-Text
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Welz, B.; Bikker, R.; Junemann, J.; Christmann, M.; Neumann, K.; Weber, M.; Hoffmeister, L.; Preuß, K.; Pich, A.; Huber, R.; Brand, K. Proteome and Phosphoproteome Analysis in TNF Long Term-Exposed Primary Human Monocytes. Int. J. Mol. Sci. 2019, 20, 1241.
Welz B, Bikker R, Junemann J, Christmann M, Neumann K, Weber M, Hoffmeister L, Preuß K, Pich A, Huber R, Brand K. Proteome and Phosphoproteome Analysis in TNF Long Term-Exposed Primary Human Monocytes. International Journal of Molecular Sciences. 2019; 20(5):1241.Chicago/Turabian Style
Welz, Bastian; Bikker, Rolf; Junemann, Johannes; Christmann, Martin; Neumann, Konstantin; Weber, Mareike; Hoffmeister, Leonie; Preuß, Katharina; Pich, Andreas; Huber, René; Brand, Korbinian. 2019. "Proteome and Phosphoproteome Analysis in TNF Long Term-Exposed Primary Human Monocytes." Int. J. Mol. Sci. 20, no. 5: 1241.
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