title: 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.
abstract: 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.
authors.affiliation: Department of Medicine, Division of Clinical Immunology and Allergy, and David Geffen School of Medicine, University of California Los Angeles, CA 90095, USA.
authors.forename: Meiying
authors.initials: M
authors.lastname: Wang
authors.forename: Gary Guishan
authors.initials: GG
authors.lastname: Xiao
authors.forename: Ning
authors.initials: N
authors.lastname: Li
authors.forename: Yongming
authors.initials: Y
authors.lastname: Xie
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Loo
authors.forename: Andre E
authors.initials: AE
authors.lastname: Nel
mesh_terms: D000393:Air Pollutants / Q000009:adverse effects; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D004781:Environmental Exposure / Q000009:adverse effects; D004847:Epithelial Cells / Q000378:metabolism*; D005456:Fluorescent Dyes; D006801:Humans; D008264:Macrophages / Q000378:metabolism*; D018384:Oxidative Stress; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D015398:Signal Transduction; D001335:Vehicle Emissions / Q000633:toxicity*
chemical_list: D000393:Air Pollutants; D005456:Fluorescent Dyes; D010750:Phosphoproteins; D001335:Vehicle Emissions