Proteome and Phosphoproteome Analysis in TNF Long Term-Exposed Primary Human Monocytes.
Int J Mol Sci, 2019/3/12;20(5)
Welz B[1], Bikker R[2], Junemann J[3, 4], Christmann M[5], Neumann K[6], Weber M[7], Hoffmeister L[8], Preuß K[9], Pich A[10, 11], Huber R[12], Brand K[13]
Affiliations
PMID: 30871024DOI: 10.3390/ijms20051241
Impact factor: 6.208
Abstract
To 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.
Keywords: NF-κB; TNF long term exposure; monocytes; phosphoproteomics; proteomics
MeSH terms
CCAAT-Enhancer-Binding Protein-beta; Glycogen Synthase Kinase 3; HeLa Cells; Humans; Inflammation; Monocytes; NF-kappa B; Phosphorylation; Proteome; Signal Transduction; THP-1 Cells; Tumor Necrosis Factor-alpha
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