Fibulin-5 is a novel binding protein for extracellular superoxide dismutase

Andrew D. Nguyen, Shinichi Itoh, Viktoria Jeney, Hiromi Yanagisawa, Mitsuaki Fujimoto, Masuko Ushio-Fukai, Tohru Fukai

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

The extracellular superoxide dismutase (ecSOD) plays an important role in atherosclerosis and endothelial function by modulating levels of the superoxide anion (O2.-) in the extracellular space. Although heparan sulfate proteoglycan is an important ligand for ecSOD, little is known about other biological binding partners of ecSOD. The goal of this study was to identify novel proteins that interact with ecSOD. A yeast two-hybrid screening of a human aorta cDNA library using ecSOD as bait identified fibulin-5 as a predominant binding protein for ecSOD. Further analysis showed that the binding domain of ecSOD within fibulin-5 mapped to its C-terminal domain. In vitro pulldown assays and coimmunoprecipitation analysis further confirmed that ecSOD interacts with fibulin-5 in vitro and in vivo. Studies using fibulin-5 -/- mice indicated that fibulin-5 is required for binding of ecSOD to vascular tissue. Importantly, the decrease in tissue-bound ecSOD levels in aortas from fibulin-5-/- mice was associated with an increase in vascular O2.- levels. Furthermore, immunohistochemical analysis using ApoE-/- mice suggested a codistribution of ecSOD and fibulin-5 in atherosclerotic vessels. In summary, we provide in this study the first evidence that the ecSOD-fibulin-5 interaction is required for ecSOD binding to vascular tissues, thereby regulating vascular O2 .- levels. This interaction may represent a novel mechanism for controlling vascular redox state in the extracellular space in various cardiovascular diseases such as atherosclerosis and hypertension in which oxidative stress is increased.

Original languageEnglish (US)
Pages (from-to)1067-1074
Number of pages8
JournalCirculation research
Volume95
Issue number11
DOIs
StatePublished - Nov 26 2004

Fingerprint

Superoxide Dismutase
Carrier Proteins
Blood Vessels
Extracellular Space
fibulin
Aorta
Atherosclerosis
Heparan Sulfate Proteoglycans
Apolipoproteins E
Gene Library
Superoxides
Oxidation-Reduction
Oxidative Stress
Cardiovascular Diseases
Yeasts
Ligands
Hypertension

Keywords

  • Atherosclerosis
  • Extracellular matrix
  • Fibulin-5
  • Superoxide
  • Superoxide dismutase

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Fibulin-5 is a novel binding protein for extracellular superoxide dismutase. / Nguyen, Andrew D.; Itoh, Shinichi; Jeney, Viktoria; Yanagisawa, Hiromi; Fujimoto, Mitsuaki; Ushio-Fukai, Masuko; Fukai, Tohru.

In: Circulation research, Vol. 95, No. 11, 26.11.2004, p. 1067-1074.

Research output: Contribution to journalArticle

Nguyen, Andrew D. ; Itoh, Shinichi ; Jeney, Viktoria ; Yanagisawa, Hiromi ; Fujimoto, Mitsuaki ; Ushio-Fukai, Masuko ; Fukai, Tohru. / Fibulin-5 is a novel binding protein for extracellular superoxide dismutase. In: Circulation research. 2004 ; Vol. 95, No. 11. pp. 1067-1074.
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