Proteome and Phosphoproteome Analysis in TNF Long Term-Exposed Primary Human Monocytes

International Journal of Molecular Sciences

Martin Christmann¹  Katharina Preuß¹  René Huber¹  Leonie Hoffmeister¹  Bastian Welz¹  Rolf Bikker¹  Mareike Weber¹  Konstantin Neumann¹  Korbinian Brand¹  Andreas Pich²  Johannes Junemann² 

[1] Institute of Clinical Chemistry, Hannover Medical School, 30625 Hannover, Germany;Institute of Toxicology, Hannover Medical School, 30625 Hannover, Germany;

关键词: proteomics;    phosphoproteomics;    TNF long term exposure;    monocytes;    NF-κB;   

DOI  :  10.3390/ijms20051241

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown