title: Protein carbonylation in human endothelial cells exposed to cigarette smoke extract.
abstract: Cigarette smoke is a significant independent risk factor for vascular diseases and is a leading cause of structural and functional alterations of the vascular endothelium. In this study, we show protein carbonylation in the human umbilical vein endothelial cell line (ECV-304) exposed to whole-phase cigarette smoke extract. The main carbonylated proteins, including cytoskeletal proteins, glycolytic enzymes, xenobiotic metabolizing and antioxidant enzymes, and endoplasmic reticulum proteins, were identified by means of two-dimensional electrophoresis and Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) mass spectrometry (redox proteomics). Morphological analyses by fluorescence microscopy evidenced alterations in the microtubule cytoskeleton, especially at longer exposure time to cigarette smoke extract. Morphological analyses by transmission electron microscopy showed vacuolisation of the cytoplasm, alteration of mitochondria ultrastructure, and some enlargement of the perinuclear space. The possible role played by protein carbonylation caused by reactive species contained in cigarette smoke in the cigarette smoke-induced endothelial injury is discussed.
authors: Department of Biotechnology and Life Sciences, University of Insubria, I-21100 Varese, Italy.
authors: Rosalba
authors: R
authors: Gornati
authors: Graziano
authors: G
authors: Colombo
authors: Marco
authors: M
authors: Clerici
authors: Federica
authors: F
authors: Rossi
authors: Nicoletta
authors: N
authors: Gagliano
authors: Consuelo
authors: C
authors: Riva
authors: Roberto
authors: R
authors: Colombo
authors: Isabella
authors: I
authors: Dalle-Donne
authors: Giovanni
authors: G
authors: Bernardini
authors: Aldo
authors: A
authors: Milzani
mesh_terms: D000975:Antioxidants / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000473:pathology; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000473:pathology; D006801:Humans; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000737:chemistry; D010084:Oxidation-Reduction; D050050:Protein Carbonylation; D011506:Proteins / Q000378:metabolism; D040901:Proteomics; D017382:Reactive Oxygen Species; D012906:Smoke / Q000009:adverse effects*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014026:Nicotiana / Q000737:chemistry / Q000633:toxicity*
chemical_list: D000975:Antioxidants; D011506:Proteins; D017382:Reactive Oxygen Species; D012906:Smoke