Heat shock protein 90 modulates endothelial nitric oxide synthase activity and vascular reactivity in the newborn piglet pulmonary circulation

Judy L. Aschner, Susan L. Foster, Mark Kaplowitz, Yongmei Zhang, Heng Zeng, Candice D. Fike

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Heat shock protein 90 (Hsp90) binding to endothelial nitric oxide synthase (eNOS) is an important step in eNOS activation. The conformational state of bound Hsp90 determines whether eNOS produces nitric oxide (NO) or superoxide (O2•-). We determined the effects of the Hsp90 antagonists geldanamycin (GA) and radicicol (RA) on basal and ACh-stimulated changes in vessel diameter, cGMP production, and Hsp90:eNOS coimmunoprecipitation in piglet resistance level pulmonary arteries (PRA). In perfused piglet lungs, we evaluated the effects of GA and RA on ACh-stimulated changes in pulmonary arterial pressure (Ppa) and perfusate accumulation of stable NO metabolites (NOx-). The effects of GA and RA on ACh-stimulated O2•- generation was investigated in cultured pulmonary microvascular endothelial cells (PMVEC) by dihydroethidine (DHE) oxidation and confocal microscopy. Hsp90 inhibition with GA or RA reduced ACh-mediated dilation, abolished the ACh-stimulated increase in cGMP, and reduced eNOS:Hsp90 coprecipitation. GA and RA also inhibited the ACh-mediated changes in Ppa and NOx- accumulation rates in perfused lungs. ACh increased the rate of DHE oxidation in PMVEC pretreated with GA and RA but not in untreated cells. The cell-permeable superoxide dismutase mimetic M40401 reversed GA-mediated inhibition of ACh-induced dilation in PRA. We conclude that Hsp90 is a modulator of eNOS activity and vascular reactivity in the newborn piglet pulmonary circulation. Uncoupling of eNOS with GA or RA inhibits ACh-mediated dilation by a mechanism that involves O2 •- generation.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume292
Issue number6
DOIs
StatePublished - Jun 2007
Externally publishedYes

Fingerprint

HSP90 Heat-Shock Proteins
Pulmonary Circulation
Nitric Oxide Synthase Type III
Blood Vessels
Lung
Dilatation
Pulmonary Artery
Nitric Oxide
Endothelial Cells
geldanamycin
monorden
Protein Binding
Confocal Microscopy
Superoxides
Superoxide Dismutase
Arterial Pressure

Keywords

  • Geldanamycin
  • Isolated perfused lungs
  • Pulmonary resistance arteries
  • Radicicol
  • Superoxide

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology

Cite this

Heat shock protein 90 modulates endothelial nitric oxide synthase activity and vascular reactivity in the newborn piglet pulmonary circulation. / Aschner, Judy L.; Foster, Susan L.; Kaplowitz, Mark; Zhang, Yongmei; Zeng, Heng; Fike, Candice D.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 292, No. 6, 06.2007.

Research output: Contribution to journalArticle

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