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Am J Physiol Heart Circ Physiol 281: H1657-H1666, 2001;
0363-6135/01 $5.00
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Vol. 281, Issue 4, H1657-H1666, October 2001

Evidence supporting the Nucleotide Axis Hypothesis: ATP release and metabolism by coronary endothelium

Iain L. O. Buxton, Robert A. Kaiser, Brian C. Oxhorn, and Dennis J. Cheek

Department of Pharmacology, University of Nevada School of Medicine, Reno, Nevada 89557

The Nucleotide Axis Hypothesis, defined and supported herein, proposes that ATP stimulates the release of vasoactive mediators from endothelium, including ATP itself. Here, we show rapid endothelium-dependent, agonist-stimulated ATP elaboration in coronary vessels of guinea pigs. Measurement of extracellular ADP metabolism in intact vessels results in the time- and substrate-dependent formation of ATP in the coronary perfusate in amounts greater than can be accounted for by release from endothelium alone. ATP formation by endothelial cells is saturable (KM = 38.5 µmol/l, where KM is substrate concentration at which rate is half-maximal.) and trypsin-sensitive, membranes from [gamma -32P]ATP-labeled cells support ADP-dependent transphosphorylation by a 20-kDa protein, Western blots reveal the presence of a nucleoside diphosphate kinase (NDPK) of ~20 kDa in endothelial membranes, and analysis of NDPK antibody binding by flow cytometry is consistent with the presence of an ecto-NDPK on cardiac endothelial cells. Sequencing of the endothelial cell ecto-NDPK reveals a predicted amino acid sequence with 85% identity to human Nm23-H1 and consistent with a protein whose properties may confer membrane association as well as sites of regulation of activity. Our data underscore the potential importance of a nucleotide axis in cardiac blood vessels.

release of vasoactive factors; ectoenzyme; ecto-Nm23


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