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Nitric oxide decreases the sensitivity of pulmonary endothelial cells to LPS-induced apoptosis in a zinc-dependent fashion

  • Zi-Lue Tang
  • Karla J. Wasserloos
  • XiangHong Liu
  • Molly S. Stitt
  • Ian J. Reyolds
  • Bruce R. Pitt
  • Claudette M. St. Croix
Part of the Developments in Molecular and Cellular Biochemistry book series (DMCB, volume 37)

Abstract

We hypothesized that: (a) S-nitrosylation of metallothionein (MT) is a component of pulmonary endothelial cell nitric oxide (NO) signaling that is associated with an increase in labile zinc; and (b) NO mediated increases in labile zinc in turn reduce the sensitivity of pulmonary endothelium to LPS-induced apoptosis. We used microspectrofluorometric techniques to show that exposing mouse lung endothelial cells (MLEC) to the NO-donor, S-nitrosocysteine, resulted in a 45% increase in fluorescence of the Zn2+-specific fluorophore, Zinquin, that was rapidly reversed by exposure to the Zn2+ chelator, NNN‘N’-tetrakis-(2pyridylmethyl)ethylenediamine; TPEN). The absence of a NO-mediated increase in labile Zn2+ in MLEC from MT-I and -II knockout mice inferred a critical role for MT in the regulation of Zn2+ homeostasis by NO. Furthermore, we found that prior exposure of cultured endothelial cells from sheep pulmonary artery (SPAEC), to the NO-donor, S-nitroso-N-acetylpenicillamine (SNAP) reduced their sensitivity to lipopolysaccharide (LPS) induced apoptosis. The anti-apoptotic effects of NO were significantly inhibited by Zn2+ chelation with low doses of TPEN (1011M). Collectively, these data suggest that S-nitrosylation of MT is associated with an increase in labile (TPEN chelatable) zinc and NO-mediated MT dependent zinc release is associated with reduced sensitivity to LPS-induced apoptosis in pulmonary endothelium. (Mol Cell Biochem 234/235: 211–217, 2002)

Key words

live cell imaging Zinquin S-nitrosylation 

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Copyright information

© Springer Science+Business Media New York 2002

Authors and Affiliations

  • Zi-Lue Tang
    • 1
  • Karla J. Wasserloos
    • 1
  • XiangHong Liu
    • 1
  • Molly S. Stitt
    • 1
  • Ian J. Reyolds
    • 2
  • Bruce R. Pitt
    • 1
  • Claudette M. St. Croix
    • 1
  1. 1.Department of Environmental and Occupational HealthThe Graduate School of Public Health, University of PittsburghPittsburghUSA
  2. 2.Department of PharmacologyUniversity of Pittsburgh School of MedicinePittsburghUSA

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