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, Kwanghoon Lee

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

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 (H2O 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 2O2 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 H2O2, 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 H2O2. These results could provide a framework for the understanding of the mechanisms of cellular redox homeostasis and H 2O2 metabolism in microendothelium environment in various biological processes as well as pathological conditions.

Original languageEnglish
Pages (from-to)1507-1519
Number of pages13
JournalProteomics
Volume5
Issue number6
DOIs
Publication statusPublished - 2005 Apr 1

Fingerprint

Endothelial cells
Proteomics
Hydrogen Peroxide
Endothelial Cells
Skin
Reactive Oxygen Species
Proteins
Metabolism
Biological Phenomena
Protein Databases
Poisons
Ultraviolet spectrophotometers
Electrophoresis, Gel, Two-Dimensional
Oxidative stress
Microvessels
Oxidation-Reduction
Electrophoresis
Mass Spectrometry
Oxidative Stress
Lasers

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Ha, Moon Kyung ; Chung, Kee Yang ; Bang, Dongsik ; Park, Yoon Kee ; Lee, Kwanghoon. / Proteomic analysis of the proteins expressed by hydrogen peroxide treated cultured human dermal microvascular endothelial cells. In: Proteomics. 2005 ; Vol. 5, No. 6. pp. 1507-1519.
@article{b746fb981c494abdabae3cfe5323cd6c,
title = "Proteomic analysis of the proteins expressed by hydrogen peroxide treated cultured human dermal microvascular endothelial cells",
abstract = "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 (H2O 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 2O2 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 H2O2, 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 H2O2. These results could provide a framework for the understanding of the mechanisms of cellular redox homeostasis and H 2O2 metabolism in microendothelium environment in various biological processes as well as pathological conditions.",
author = "Ha, {Moon Kyung} and Chung, {Kee Yang} and Dongsik Bang and Park, {Yoon Kee} and Kwanghoon Lee",
year = "2005",
month = "4",
day = "1",
doi = "10.1002/pmic.200401043",
language = "English",
volume = "5",
pages = "1507--1519",
journal = "Proteomics",
issn = "1615-9853",
publisher = "Wiley-VCH Verlag",
number = "6",

}

Proteomic analysis of the proteins expressed by hydrogen peroxide treated cultured human dermal microvascular endothelial cells. / Ha, Moon Kyung; Chung, Kee Yang; Bang, Dongsik; Park, Yoon Kee; Lee, Kwanghoon.

In: Proteomics, Vol. 5, No. 6, 01.04.2005, p. 1507-1519.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Proteomic analysis of the proteins expressed by hydrogen peroxide treated cultured human dermal microvascular endothelial cells

AU - Ha, Moon Kyung

AU - Chung, Kee Yang

AU - Bang, Dongsik

AU - Park, Yoon Kee

AU - Lee, Kwanghoon

PY - 2005/4/1

Y1 - 2005/4/1

N2 - 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 (H2O 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 2O2 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 H2O2, 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 H2O2. These results could provide a framework for the understanding of the mechanisms of cellular redox homeostasis and H 2O2 metabolism in microendothelium environment in various biological processes as well as pathological conditions.

AB - 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 (H2O 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 2O2 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 H2O2, 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 H2O2. These results could provide a framework for the understanding of the mechanisms of cellular redox homeostasis and H 2O2 metabolism in microendothelium environment in various biological processes as well as pathological conditions.

UR - http://www.scopus.com/inward/record.url?scp=17844364237&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=17844364237&partnerID=8YFLogxK

U2 - 10.1002/pmic.200401043

DO - 10.1002/pmic.200401043

M3 - Article

C2 - 15838903

AN - SCOPUS:17844364237

VL - 5

SP - 1507

EP - 1519

JO - Proteomics

JF - Proteomics

SN - 1615-9853

IS - 6

ER -