identification of a tsetse fly salivary protein with dual inhibitory action on human platelet aggregation采采蝇的识别唾液蛋白质与双人类血小板聚集抑制作用.pdfVIP
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identification of a tsetse fly salivary protein with dual inhibitory action on human platelet aggregation采采蝇的识别唾液蛋白质与双人类血小板聚集抑制作用
Identification of a Tsetse Fly Salivary Protein with Dual Inhibitory Action on Human Platelet Aggregation 1,2,3 1 2,3 1 1 Guy Caljon , Karin De Ridder , Patrick De Baetselier , Marc Coosemans , Jan Van Den Abbeele * 1 Unit of Entomology, Institute of Tropical Medicine Antwerp (ITM), Antwerp, Belgium, 2 Unit of Cellular and Molecular Immunology, Vrije Universiteit Brussel (VUB), Brussels, Belgium, 3 Department of Molecular and Cellular Interactions, Vlaams Instituut voor Biotechnologie (VIB), Ghent, Belgium Abstract Background: Tsetse flies (Glossina sp.), the African trypanosome vectors, rely on anti-hemostatic compounds for efficient blood feeding. Despite their medical importance, very few salivary proteins have been characterized and functionally annotated. Methodology/Principal Findings: Here we report on the functional characterisation of a 59nucleotidase-related (59Nuc) saliva protein of the tsetse fly Glossina morsitans morsitans. This protein is encoded by a 1668 bp cDNA corresponding at the genomic level with a single-copy 4 kb gene that is exclusively transcribed in the tsetse salivary gland tissue. The encoded 59Nuc protein is a soluble 65 kDa glycosylated compound of tsetse saliva with a dual anti-hemostatic action that relies on its combined apyrase activity and fibrinogen receptor (GPIIb/IIIa) antagonistic properties. Experimental evidence is based on the biochemical and functional characterization of recombinant protein and on the successful silencing of the 59nuc translation in the salivary gland by RNA interference (RNAi). Refolding of a 5 9Nuc/SUMO-fusion protein yielded an active apyrase enzyme with Km and Vmax values of 43 64 mM
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