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Proteomic Analysis of Vascular Smooth Muscle Cells Treated With Ouabain

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Cardiovascular Proteomics

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 357))

Abstract

Apoptosis of vascular smooth muscle cells (VSMC) plays an important role in remodeling the vessel walls, one of the major determinants of long-term blood pressure elevation and an independent risk factor for cardiovascular morbidity and mortality. Apoptosis in VSMC can be inhibited by inversion of the intracellular [Na+]/[K+] ratio after the sustained blockage of the Na+,K+-ATPase by ouabain. Using two-dimensional gel electrophoresis followed by tandem mass spectroscopy, we compared proteomes of control VSMC and of those with ouabain-inhibited Na+,K+-ATPase and found that ouabain treatment led to overexpression of numerous soluble and membrane-bound proteins. Among proteins, which showed the highest level of ouabain-induced expression, we identified mortalin (also known as GRP75 or PBP-74), a member of the heat shock protein 70 superfamily and a marker for cellular mortal and immortal phenotypes. Further experiments showed that mortalin RNA and protein levels are induced in ouabain-treated VSMC, and that transient transfection of cells with mortalin cDNA inhibited serum deprivation-induced apoptosis via inactivation of the tumor suppressor gene, p53.

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Author information

Authors and Affiliations

  1. Department of Medical Genetics, St. Justine Hospital, Côte St. Catherine, Montreal, Quebec, Canada

    Alexey V. Pshezhetsky

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  1. Alexey V. Pshezhetsky
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Editors and Affiliations

  1. Department of Immunology, Fundación Jiménez Díaz, Universidad Complutense, Madrid, Spain

    Fernando Vivanco

  2. Department of Biochemistry and Molecular Biology I, Universidad Complutense, Madrid, Spain

    Fernando Vivanco

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© 2007 Humana Press Inc.

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Pshezhetsky, A.V. (2007). Proteomic Analysis of Vascular Smooth Muscle Cells Treated With Ouabain. In: Vivanco, F. (eds) Cardiovascular Proteomics. Methods in Molecular Biology™, vol 357. Humana Press. https://doi.org/10.1385/1-59745-214-9:253

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  • DOI: https://doi.org/10.1385/1-59745-214-9:253

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-535-4

  • Online ISBN: 978-1-59745-214-4

  • eBook Packages: Springer Protocols

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