Elsevier

Biochemical Pharmacology

Volume 39, Issue 1, 1 January 1990, Pages 75-84
Biochemical Pharmacology

Characterization of cyclic nucleotide phosphodiesterases from cultured bovine aortic endothelial cells

https://doi.org/10.1016/0006-2952(90)90650-AGet rights and content

Abstract

Experiments were carried out in order to isolate and characterize the cyclic nucleotide phosphodiesterase activities in primary and low passages of cultured bovine aortic endothelial cells. The subcellular characterization of the cyclic nucleotide hydrolytic activity showed that both cAMP and cGMP hydrolytic activities were predominant in the cytosolic rather than the particulate fraction of the endothelial cell homogenate. At a low substrate concentration (0.25 μM), the major hydrolytic activity was for cAMP while at a high concentration (20 μM) it was for both cAMP and cGMP. Both cAMP and cGMP hydrolytic activities were insensitive to calmodulin. Cytosolic cyclic nucleotide phosphodiesterase activity was resolved into two distinct phosphodiesterase forms using HPLC. The first eluted form was designated cGS-PDE: it hydrolysed both cAMP and cGMP and its cAMP hydrolytic activity was markedly enhanced by the presence of cGMP. The second form was designated cAMP-PDE: it selectively hydrolysed cAMP. The cytosolic cAMP-PDE was inhibited by micromolar concentrations of cAMP-PDE inhibitors such as trequinsin, rolipram, dipyridamole or papaverine. The cGS-PDE was inhibited by micromolar concentrations of trequinsin, dipyridamole and papaverine and was insensitive to rolipram, except for the hydrolysis of cAMP which was inhibited in the micromolar range. Both the cAMP-PDE and the cGS-PDE were relatively insensitive to the selective cGMP-PDE inhibitor, zaprinast which was about 750-fold less potent on endothelial PDEs than on smooth muscle cGMP-PDE. The identification of selective and specific PDE inhibitors of the different PDE forms may allow a better understanding of the regulation and the role of cyclic nucleotides in endothelial cells.

References (31)

  • RF O'Brien et al.

    Endothelial cells in culture produce a vasoconstrictor substance

    J Cell Physiol

    (1987)
  • M Yanagisawa et al.

    A novel potent vasoconstrictor peptide produced by vascular endothelial cells

    Nature (Lond)

    (1988)
  • V Buonassisi et al.

    Hormone and neurotransmitter receptor in an established vascular endothelial cell line

  • JS Makarski

    Stimulation of cAMP production by vasoactive agents in cultured bovine aortic and pulmonary artery endothelial cells

    In Vitro

    (1981)
  • NK Hopkins et al.

    Regulation of endothelial cell cyclic nucleotide metabolism by prostacyclin

    J Clin Invest

    (1981)
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    Present address: Baylor College of Medicine, Center for Experimental Therapeutics, One Baylor Plaza, Houston, TX 77030, U.S.A.

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