Inhibition of platelet function by recombinant soluble ecto-ADPase/CD39.

R B Gayle; C R Maliszewski; S D Gimpel; M A Schoenborn; R G Caspary; C Richards; K Brasel; V Price; J H Drosopoulos; N Islam; T N Alyonycheva; M J Broekman; A J Marcus

doi:10.1172/JCI1753

Inhibition of platelet function by recombinant soluble ecto-ADPase/CD39.

R B Gayle 3rd, C R Maliszewski, S D Gimpel, M A Schoenborn, R G Caspary, C Richards, K Brasel, V Price, J H Drosopoulos, N Islam, T N Alyonycheva, M J Broekman, and A J Marcus

Published May 1, 1998 - More info

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Abstract

Excessive platelet accumulation and recruitment, leading to vessel occlusion at sites of vascular injury, present major therapeutic challenges in cardiovascular medicine. Endothelial cell CD39, an ecto-enzyme with ADPase and ATPase activities, rapidly metabolizes ATP and ADP released from activated platelets, thereby abolishing recruitment. Therefore, a soluble form of CD39, retaining nucleotidase activities, would constitute a novel antithrombotic agent. We designed a recombinant, soluble form of human CD39, and isolated it from conditioned media from transiently transfected COS-1 cells and from stably transfected Chinese hamster ovary (CHO) cells. Conditioned medium from CHO cells grown under serum-free conditions was subjected to anti-CD39 immunoaffinity column chromatography, yielding a single approximately 66-kD protein with ATPase and ADPase activities. Purified soluble CD39 blocked ADP-induced platelet aggregation in vitro, and inhibited collagen-induced platelet reactivity. Kinetic analyses indicated that, while soluble CD39 had a Km for ADP of 5.9 microM and for ATP of 2.1 microM, the specificity constant kcat/Km was the same for both substrates. Intravenously administered soluble CD39 remained active in mice for an extended period of time, with an elimination phase half-life of almost 2 d. The data indicate that soluble CD39 is a potential therapeutic agent for inhibition of platelet-mediated thrombotic diatheses.

Inhibition of platelet function by recombinant soluble ecto-ADPase/CD39.

R B Gayle 3rd, C R Maliszewski, S D Gimpel, M A Schoenborn, R G Caspary, C Richards, K Brasel, V Price, J H Drosopoulos, N Islam, T N Alyonycheva, M J Broekman, and A J Marcus

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Inhibition of platelet function by recombinant soluble ecto-ADPase/CD39.

R B Gayle 3rd, C R Maliszewski, S D Gimpel, M A Schoenborn, R G Caspary, C Richards, K Brasel, V Price, J H Drosopoulos, N Islam, T N Alyonycheva, M J Broekman, and A J Marcus

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