Ecto-ATPases of clinical and non-clinical isolates of Acanthamoeba.
Acanthamoeba are opportunistic protozoan parasites that can cause fatal granulomatous amoebic encephalitis and eye keratitis, however the pathogenic mechanisms of Acanthamoeba remain unclear. In this study, we described the ability of live Acanthamoeba to hydrolyse extracellular ATP. Both clinical and non-clinical isolates belonging to genotypes, T1, T2, T3, T4 and T7 exhibited ecto-ATPase activities in vitro. Using non-denaturing polyacrylamide gel electrophoresis, ecto-ATPases were further characterized. All Acanthamoeba isolates tested, exhibited a single ecto-ATPase band (approximate molecular weight of 272 kDa). However, clinical isolates exhibited additional bands suggesting that ecto-ATPases may play a role in the pathogenesis of Acanthamoeba. This was supported using suramin (ecto-ATPase inhibitor), which inhibited Acanthamoeba-induced host cell cytotoxicity. Previously, we and others have shown that Acanthamoeba binds to host cells using their mannose-binding protein and binding can be blocked using exogenous alpha-mannose. In this study, we observed that alpha-mannose significantly increased ecto-ATPase activities of pathogenic Acanthamoeba belonging to T1, T2, T3 and T4 genotypes but had no effect on non-pathogenic Acanthamoeba (belonging to T7 genotype). Overall, we have shown, for the first time, that Acanthamoeba exhibit ecto-ATPase activities, which may play a role in the pathogenesis of Acanthamoeba as well as their potential role in the differentiation of pathogenic Acanthamoeba.
Sissons
James
J
School of Biological and Chemical Sciences, Birkbeck College, University of London, London WC1E 7HX, UK.
Alsam
Selwa
S
Jayasekera
Samantha
S
Khan
Naveed Ahmed
NA
D000048:Acanthamoeba / Q000201:enzymology* / Q000235:genetics; D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002448:Cell Adhesion; D004847:Epithelial Cells / Q000166:cytology / Q000469:parasitology; D019573:Epithelium, Corneal / Q000166:cytology / Q000469:parasitology; D005786:Gene Expression Regulation; D006801:Humans; D006868:Hydrolysis; D008358:Mannose / Q000378:metabolism; D037601:Mannose-Binding Lectin / Q000378:metabolism; D013498:Suramin / Q000494:pharmacology
D037601:Mannose-Binding Lectin; D013498:Suramin; D000251:Adenosine Triphosphatases; C027201:ectoATPase; D008358:Mannose