Expression of functional recombinant mosquito salivary apyrase: a potential therapeutic platelet aggregation inhibitor.
Autor(es): Sun Dongfeng; McNicol Archibald; James Anthony A; Peng Zhikang
Resumo: Excessive platelet activation and accumulation can lead to vessel occlusion and thus present major therapeutic challenges in cardiovascular medicine. Apyrase, an ecto-enzyme with ADPase and ATPase activities, rapidly metabolizes ADP and ATP released from platelets and endothelial cells, thereby reducing platelet activation and recruitment. In the present study, we expressed a 68-kDa recombinant mosquito (Aedes aegypti) salivary apyrase using a baculovirus/insect cell expression system and purified it to homogeneity using anion-exchange chromatography on a large scale. A yield of 18 mg of purified recombinant apyrase was obtained from 1 litre of the medium. Kinetic analysis indicated that the recombinant apyrase had a K(m) of 12.5 microM for ADP and a K(m) of 15.0 microM for ATP. The recombinant apyrase inhibited ADP-, collagen- and thrombin-induced human platelet aggregation in a dose-dependent manner, indicating that the recombinant protein retained nucleotidase activity in a whole cell system, which suggests that it may serve as a therapeutic agent for inhibition of platelet-mediated thrombosis.
Imprenta: Platelets, v. 17, n. 3, p. 178-184, 2009
Identificador do objeto digital: https://doi.org/10.1080/09537100500460234
Descritores: Aedes aegypti - Cell ; Aedes aegypti - Pathogenesis ; Aedes aegypti - Protein synthesis ; Aedes aegypti - Proteins ; Aedes aegypti - virus
Data de publicação: 2006
