Inhibition of platelet function by recombinant soluble Ecto-ADPase/CD39

162Citations
Citations of this article
44Readers
Mendeley users who have this article in their library.

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 ~ 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 K(m) for ADP of 5.9 μM and for ATP of 2.1 μM, the specificity constant k(cat)/K(m) 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.

Cite

CITATION STYLE

APA

Gayle, R. B., Maliszewski, C. R., Gimpel, S. D., Schoenborn, M. A., Caspary, R. G., Richards, C., … Marcus, A. J. (1998). Inhibition of platelet function by recombinant soluble Ecto-ADPase/CD39. Journal of Clinical Investigation, 101(9), 1851–1859. https://doi.org/10.1172/JCI1753

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free