Identification of nuclear phosphoproteins as novel tobacco markers in mouse lung tissue following short-term exposure to tobacco smoke

摘要

class="kwd-title">Abbreviations: COPD, chronic obstructive pulmonary disorder; FACTp140, FACT complex subunit SPT16; K18, keratin type 1 cytoskeletal 18; AFABP, adipocyte fatty acid-binding protein; OSF3, peroxiredoxin-1; SPTBN1, spectrin β chain brain 1; STAT, signal transducer and activator of transcription; IL, interleukin; K8, keratin type 2 cytoskeletal 8; PRP19, pre-mRNA-processing factor 19; CRP1, cysteine and glycine-rich protein 1; Jak2, tyrosine-protein kinase JAK2; pSTAT3-Tyr705, phosphorylated STAT3; TIM, mitochondrial import inner membrane translocase subunit Tim9; HIP1, Huntingtin-interacting protein 1; 60s-RP, 60s ribosomal protein L10E; TNF, tumor necrosis factor; ALDH2, aldehyde dehydrogenase, mitochondrial; PRP19, pre-mRNA-processing factor 19; ROS, reactive oxygen species; TNFR2, tumor necrosis factor receptor 2; JNK, c-Jun NH2-terminal kinase; ERK(1/2), extracellular signal regulated kinase 1/2; NF-κB, nuclear factor-kappa B; PKC-α, protein kinase C-α; p100, serine protease P100; MAPK3, mitogen-activated protein kinase 3; TGF-β, Transforming growth factor-β; TRAP1, heat shock protein 75 kDa; LIM, LIM/homeobox protein class="kwd-title">Keywords: Tobacco smoke exposure, Nuclear phosphoprotein, Phosphoproteomic analysis, Signaling pathways class="head no_bottom_margin" id="idm140160846035776title">AbstractSmoking is a risk factor for lung diseases, including chronic obstructive pulmonary disease and lung cancer. However, the molecular mechanisms mediating the progression of these diseases remain unclear. Therefore, we sought to identify signaling pathways activated by tobacco-smoke exposure, by analyzing nuclear phosphoprotein expression using phosphoproteomic analysis of lung tissue from mice exposed to tobacco smoke. Sixteen mice were exposed to tobacco smoke for 1 or 7 days, and the expression of phosphorylated peptides was analyzed by mass spectrometry. A total of 253 phosphoproteins were identified, including FACT complex subunit SPT16 in the 1-day exposure group, keratin type 1 cytoskeletal 18 (K18), and adipocyte fatty acid-binding protein, in the 7-day exposure group, and peroxiredoxin-1 (OSF3) and spectrin β chain brain 1 (SPTBN1), in both groups. Semi-quantitative analysis of the identified phosphoproteins revealed that 33 proteins were significantly differentially expressed between the control and exposed groups. The identified phosphoproteins were classified according to their biological functions. We found that the identified proteins were related to inflammation, regeneration, repair, proliferation, differentiation, morphogenesis, and response to stress and nicotine. In conclusion, we identified proteins, including OSF3 and SPTBN1, as candidate tobacco smoke-exposure markers; our results provide insights into the mechanisms of tobacco smoke-induced diseases.