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Use of a fluorescent phosphoprotein dye to characterize oxidative stress‐induced signaling pathway components in macrophage and epithelial cultures exposed to diesel exhaust particle chemicals
Author(s) -
Wang Meiying,
Xiao Gary Guishan,
Li Ning,
Xie Yongming,
Loo Joseph A.,
Nel Andre E.
Publication year - 2005
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200410428
Subject(s) - oxidative stress , signal transduction , microbiology and biotechnology , chemistry , oxidative phosphorylation , inflammation , diesel exhaust , biochemistry , biology , immunology , diesel fuel , organic chemistry
A large body of evidence has shown that exposure to ambient particulate matter (PM) leads to asthma exacerbation through an excitation of allergic inflammation. Utilizing diesel exhaust particles (DEPs) as a model air pollutant, we and others have demonstrated that PM contains redox‐active chemicals that generate inflammation through an oxidative stress mechanism. Recently, the strengths of proteomics have enabled us to demonstrate that organic DEP extracts induce a hierarchical expression pattern of oxidative stress‐induced proteins in macrophages and epithelial cells. As a further extension of this work, we now employ a new phosphosensor fluorescent dye, Pro‐Q Diamond, to elucidate the induction of phosphoproteins and intracellular signaling cascades that may play a role in DEP‐induced inflammation. We demonstrate that DEPs induced the phosphorylation of several phosphoproteins that belong to a number of signaling pathways as well as other oxidative stress pathways. In combination with cytokine array, phosphoproteome analysis using Pro‐Q Diamond allowed us to characterize the aromatic and polar chemicals of DEPs that are involved in the activation of three different mitogen‐activated protein (MAP) kinase signaling pathways.