Oxidative modifications of proteins by sodium arsenite in human umbilical vein endothelial cells.
Epidemiologic studies have demonstrated that chronic arsenic exposure is associated with the incidence of chronic diseases. This association is partly related to the increase in reactive oxygen species (ROS) overload and protein oxidation that result from arsenic exposure. In this study, we intended to identify proteins susceptible to oxidative carbonylation by sodium arsenite and the impact of carbonylation on the function of these proteins in human umbilical vein endothelial cells (HUVECs). The 2,4-dinitrophenylhydrazine (DNPH) dot-blot assay revealed that arsenite (0-50 μM) dose-dependently increased protein carbonylation. Consistent with these findings, the cellular ROS level as measured by 2',7'-dichlorofluorescein diacetate (DCHF-DA) assay was increased in cells exposed to arsenite. By two-dimensional gel electrophoresis and matrix assist laser desorption ionization time of flight mass spectrometry (MALDI-TOF/MS), one glycolytic enzyme, enolase-α, two cytoskeleton proteins, fascin (F-actin associated protein) and vimentin, and two protein quality control proteins, HSC70 (heat-shock cognate protein 70), and PDIA3 (protein disulfide isomerase family A, member 3) were identified to be arsenic-sensitive carbonlyated proteins. Accompanied by carbonylation, enolase-α activity was dose-dependently decreased and the F-actin filament network was disturbed. Taken together, our results suggest that arsenite exposure results in the generation of carbonylated proteins, and the resultant changes in energy metabolism and in the cytoskeletal network may partly lead to cell damage.
Lii
Chong-Kuei
CK
Department of Nutrition, China Medical University, Taichung, Taiwan.
Lin
Ai-Hsuan
AH
Lee
Shu-Lien
SL
Chen
Haw-Wen
HW
Wang
Tsu-Shing
TS
D018053:Arsenites / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D010084:Oxidation-Reduction; D050050:Protein Carbonylation; D011506:Proteins / Q000737:chemistry*; D017382:Reactive Oxygen Species / Q000378:metabolism; D017670:Sodium Compounds / Q000494:pharmacology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D018053:Arsenites; D011506:Proteins; D017382:Reactive Oxygen Species; D017670:Sodium Compounds; C017947:sodium arsenite