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Peroxynitrite modifies the structure and function of the extracellular matrix proteoglycan perlecan by reaction with both the protein core and the heparan sulfate chains
- Source :
- Free Radical Biology & Medicine
- Publisher :
- Elsevier Inc.
-
Abstract
- The heparan sulfate (HS) proteoglycan perlecan is a major component of basement membranes, plays a key role in extracellular matrix (ECM) structure, interacts with growth factors and adhesion molecules, and regulates the adhesion, differentiation and proliferation of vascular cells. Atherosclerosis is characterized by chronic inflammation and the presence of oxidized materials within lesions, with the majority of protein damage present on ECM, rather than cell, proteins. Weakening of ECM structure plays a key role in lesion rupture, the major cause of heart attacks and strokes. In this study peroxynitrite, a putative lesion oxidant, is shown to damage perlecan structurally and functionally. Exposure of human perlecan to peroxynitrite decreases recognition by antibodies raised against both the core protein and heparan sulfate chains; dose-dependent formation of 3-nitrotyrosine was also detected. These effects were modulated by bicarbonate and reaction pH. Oxidant exposure resulted in aggregate formation, consistent with oxidative protein crosslinking. Peroxynitrite treatment modified functional properties of perlecan that are dependent on both the protein core (decreased binding of human coronary artery endothelial cells), and the HS chains (diminished fibroblast growth factor-2 (FGF-2) receptor-mediated proliferation of Baf-32 cells). The latter is consistent with a decrease in FGF-2 binding to the HS chains of modified perlecan. Immunofluorescence of advanced human atherosclerotic lesions provided evidence for the presence of perlecan and extensive formation of 3-nitrotyrosine epitopes within the intimal region; these materials showing marked co-localization. These data indicate that peroxynitrite induces major structural and functional changes to perlecan and that damage to this material occurs within human atherosclerotic lesions.
- Subjects :
- ABTS, 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)
MTT, 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan
Coronary Artery Disease
3-nitroTyr, 3-nitrotyrosine
Biochemistry
Peroxynitrite
Extracellular matrix
chemistry.chemical_compound
ONOO-, peroxynitrous acid anion
FGF-2, fibroblast growth factor 2
Cell proliferation
0303 health sciences
biology
Chemistry
Cell adhesion molecule
030302 biochemistry & molecular biology
Heparan sulfate
Original Contribution
Coronary Vessels
Immunohistochemistry
Cell biology
ECM, extracellular matrix
medicine.anatomical_structure
HCAEC, human coronary artery endothelial cells
Heparan sulfate proteoglycans
Protein Binding
HS, heparan sulfate
ONOOH, peroxynitrous acid
Perlecan
TCA, trichloroacetic acid
Cell Line
03 medical and health sciences
Physiology (medical)
Peroxynitrous Acid
medicine
Humans
HSPG, heparan sulfate proteoglycan
Cell adhesion
Fibroblast
030304 developmental biology
Inflammation
dONOO, decomposed peroxynitrite
Plaque rupture
Epithelial Cells
Atherosclerosis
Oxidative Stress
Proteoglycan
biology.protein
Heparitin Sulfate
Protein Multimerization
Tunica Intima
Subjects
Details
- Language :
- English
- ISSN :
- 08915849
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Free Radical Biology and Medicine
- Accession number :
- edsair.doi.dedup.....d16c3f98549551acae07b936c60a2641
- Full Text :
- https://doi.org/10.1016/j.freeradbiomed.2010.04.018