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Proteomic analysis of the proteins expressed by hydrogen peroxide treated cultured human dermal microvascular endothelial cells

 Moon Kyung Ha  ;  Kee Yang Chung  ;  Dongsik Bang  ;  Yoon Kee Park  ;  Kwang Hoon Lee 
 PROTEOMICS, Vol.5(6) : 1507-1519, 2005 
Journal Title
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Amino Acid Sequence ; Cell Line ; Cells, Cultured ; Cytoplasm/metabolism ; Electrophoresis, Gel, Two-Dimensional ; Endothelial Cells/metabolism* ; Flow Cytometry ; Humans ; Hydrogen Peroxide/pharmacology* ; Microcirculation/cytology ; Microcirculation/metabolism ; Mitochondria/drug effects ; Mitochondria/metabolism ; Molecular Sequence Data ; Proteome/biosynthesis* ; Reactive Oxygen Species/metabolism ; Skin/blood supply* ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Spectrophotometry, Ultraviolet ; beta-Galactosidase/metabolism
Reactive oxygen species (ROS) have been traditionally regarded as toxic by-products of aerobic metabolism. However, ROS also act as intracellular signaling molecules and can mediate phenotypes in vascular endothelial cells, which may be physiological or pathological in nature. To clarify the molecular mechanisms of ROS signaling, we examined hydrogen peroxide (H(2)O(2))-responsive proteins in cultured human dermal microvascular endothelial cells (HMVEC) using proteomic tools. Protein expression in HMVEC was studied after they had been exposed to low- and high-levels of H(2)O(2) for various times, and intracellular ROS production was examined by flow cytometer and UV spectrophotometer. Proteins obtained from dose- and time-dependent series were separated by two-dimensional gel electrophoresis and tentatively identified by matrix-assisted laser desorption-time of flight mass spectrometry, by matching the tryptic mass maps obtained with entries in the NCBI and Swiss-Prot protein sequence database. At least 163 proteins were changed by H(2)O(2), and 60 proteins were identified. Oxidative stress triggered dramatic change in the expression of proteins in primary microvessel endothelial cells, and their mapping to cellular process provided a view of the ubiquitous cellular changes elicited by H(2)O(2). These results could provide a framework for the understanding of the mechanisms of cellular redox homeostasis and H(2)O(2) metabolism in microendothelium environment in various biological processes as well as pathological conditions.
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1. College of Medicine (의과대학) > Dept. of Dermatology (피부과학교실) > 1. Journal Papers
Yonsei Authors
Park, Yoon Kee(박윤기)
Bang, Dong Sik(방동식)
Lee, Kwang Hoon(이광훈)
Chung, Kee Yang(정기양) ORCID logo https://orcid.org/0000-0003-3257-0297
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