Acute hypoxia enhances proteins' S-nitrosylation in endothelial cells

Shih-Chung Chen, Bin Huang, Yu Chi Liu, Kou-Gi Shyu, Pen Y. Lin, Danny Ling Wang

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Hypoxia-induced responses are frequently encountered during cardiovascular injuries. Hypoxia triggers intracellular reactive oxygen species/nitric oxide (NO) imbalance. Recent studies indicate that NO-mediated S-nitrosylation (S-NO) of cysteine residue is a key posttranslational modification of proteins. We demonstrated that acute hypoxia to endothelial cells (ECs) transiently increased the NO levels via endothelial NO synthase (eNOS) activation. A modified biotin-switch method coupled with Western blot on 2-dimentional electrophoresis (2-DE) demonstrated that at least 11 major proteins have significant increase in S-NO after acute hypoxia. Mass analysis by CapLC/Q-TOF identified those as Ras-GTPase-activating protein, protein disulfide-isomerase, human elongation factor-1-delta, tyrosine 3/tryptophan 5-monooxygenase activating protein, and several cytoskeleton proteins. The S-nitrosylated cysteine residue on tropomyosin (Cys 170) and β-actin (Cys 285) was further verified with the trypsic peptides analyzed by MASCOT search program. Further understanding of the functional relevance of these S-nitrosylated proteins may provide a molecular basis for treating ischemia-induced vascular disorders.

Original languageEnglish
Pages (from-to)1274-1278
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume377
Issue number4
DOIs
Publication statusPublished - Dec 26 2008

Fingerprint

Protein S
Endothelial cells
Endothelial Cells
Nitric Oxide
Cysteine
Proteins
ras GTPase-Activating Proteins
Peptide Elongation Factor 1
Protein Disulfide-Isomerases
Tryptophan Hydroxylase
Tropomyosin
Nitric Oxide Synthase Type III
Post Translational Protein Processing
Biotin
Mixed Function Oxygenases
Electrophoresis
Cytoskeleton
Tryptophan
Blood Vessels
Actins

Keywords

  • Biotin-switch
  • Endothelial cell
  • Hypoxia
  • Nitric oxide
  • Proteomics
  • S-nitrosylation

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Acute hypoxia enhances proteins' S-nitrosylation in endothelial cells. / Chen, Shih-Chung; Huang, Bin; Liu, Yu Chi; Shyu, Kou-Gi; Lin, Pen Y.; Wang, Danny Ling.

In: Biochemical and Biophysical Research Communications, Vol. 377, No. 4, 26.12.2008, p. 1274-1278.

Research output: Contribution to journalArticle

Chen, Shih-Chung ; Huang, Bin ; Liu, Yu Chi ; Shyu, Kou-Gi ; Lin, Pen Y. ; Wang, Danny Ling. / Acute hypoxia enhances proteins' S-nitrosylation in endothelial cells. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 377, No. 4. pp. 1274-1278.
@article{c03d864768bc4193a4e2f6dabf0edbdb,
title = "Acute hypoxia enhances proteins' S-nitrosylation in endothelial cells",
abstract = "Hypoxia-induced responses are frequently encountered during cardiovascular injuries. Hypoxia triggers intracellular reactive oxygen species/nitric oxide (NO) imbalance. Recent studies indicate that NO-mediated S-nitrosylation (S-NO) of cysteine residue is a key posttranslational modification of proteins. We demonstrated that acute hypoxia to endothelial cells (ECs) transiently increased the NO levels via endothelial NO synthase (eNOS) activation. A modified biotin-switch method coupled with Western blot on 2-dimentional electrophoresis (2-DE) demonstrated that at least 11 major proteins have significant increase in S-NO after acute hypoxia. Mass analysis by CapLC/Q-TOF identified those as Ras-GTPase-activating protein, protein disulfide-isomerase, human elongation factor-1-delta, tyrosine 3/tryptophan 5-monooxygenase activating protein, and several cytoskeleton proteins. The S-nitrosylated cysteine residue on tropomyosin (Cys 170) and β-actin (Cys 285) was further verified with the trypsic peptides analyzed by MASCOT search program. Further understanding of the functional relevance of these S-nitrosylated proteins may provide a molecular basis for treating ischemia-induced vascular disorders.",
keywords = "Biotin-switch, Endothelial cell, Hypoxia, Nitric oxide, Proteomics, S-nitrosylation",
author = "Shih-Chung Chen and Bin Huang and Liu, {Yu Chi} and Kou-Gi Shyu and Lin, {Pen Y.} and Wang, {Danny Ling}",
year = "2008",
month = "12",
day = "26",
doi = "10.1016/j.bbrc.2008.10.144",
language = "English",
volume = "377",
pages = "1274--1278",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Elsevier B.V.",
number = "4",

}

TY - JOUR

T1 - Acute hypoxia enhances proteins' S-nitrosylation in endothelial cells

AU - Chen, Shih-Chung

AU - Huang, Bin

AU - Liu, Yu Chi

AU - Shyu, Kou-Gi

AU - Lin, Pen Y.

AU - Wang, Danny Ling

PY - 2008/12/26

Y1 - 2008/12/26

N2 - Hypoxia-induced responses are frequently encountered during cardiovascular injuries. Hypoxia triggers intracellular reactive oxygen species/nitric oxide (NO) imbalance. Recent studies indicate that NO-mediated S-nitrosylation (S-NO) of cysteine residue is a key posttranslational modification of proteins. We demonstrated that acute hypoxia to endothelial cells (ECs) transiently increased the NO levels via endothelial NO synthase (eNOS) activation. A modified biotin-switch method coupled with Western blot on 2-dimentional electrophoresis (2-DE) demonstrated that at least 11 major proteins have significant increase in S-NO after acute hypoxia. Mass analysis by CapLC/Q-TOF identified those as Ras-GTPase-activating protein, protein disulfide-isomerase, human elongation factor-1-delta, tyrosine 3/tryptophan 5-monooxygenase activating protein, and several cytoskeleton proteins. The S-nitrosylated cysteine residue on tropomyosin (Cys 170) and β-actin (Cys 285) was further verified with the trypsic peptides analyzed by MASCOT search program. Further understanding of the functional relevance of these S-nitrosylated proteins may provide a molecular basis for treating ischemia-induced vascular disorders.

AB - Hypoxia-induced responses are frequently encountered during cardiovascular injuries. Hypoxia triggers intracellular reactive oxygen species/nitric oxide (NO) imbalance. Recent studies indicate that NO-mediated S-nitrosylation (S-NO) of cysteine residue is a key posttranslational modification of proteins. We demonstrated that acute hypoxia to endothelial cells (ECs) transiently increased the NO levels via endothelial NO synthase (eNOS) activation. A modified biotin-switch method coupled with Western blot on 2-dimentional electrophoresis (2-DE) demonstrated that at least 11 major proteins have significant increase in S-NO after acute hypoxia. Mass analysis by CapLC/Q-TOF identified those as Ras-GTPase-activating protein, protein disulfide-isomerase, human elongation factor-1-delta, tyrosine 3/tryptophan 5-monooxygenase activating protein, and several cytoskeleton proteins. The S-nitrosylated cysteine residue on tropomyosin (Cys 170) and β-actin (Cys 285) was further verified with the trypsic peptides analyzed by MASCOT search program. Further understanding of the functional relevance of these S-nitrosylated proteins may provide a molecular basis for treating ischemia-induced vascular disorders.

KW - Biotin-switch

KW - Endothelial cell

KW - Hypoxia

KW - Nitric oxide

KW - Proteomics

KW - S-nitrosylation

UR - http://www.scopus.com/inward/record.url?scp=56349164708&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=56349164708&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2008.10.144

DO - 10.1016/j.bbrc.2008.10.144

M3 - Article

C2 - 18992711

AN - SCOPUS:56349164708

VL - 377

SP - 1274

EP - 1278

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 4

ER -