Document Detail


Biochemical characterization and functional studies of Acanthamoeba mannose-binding protein.
MedLine Citation:
PMID:  16113295     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Acanthamoebae produce a painful, sight-threatening corneal infection. The adhesion of parasites to the host cells is a critical first step in the pathogenesis of infection. Subsequent to adhesion, the parasites produce a potent cytopathic effect (CPE) leading to target cell death. Recent studies showing that acanthamoebae express a mannose-binding protein (MBP) and that free alpha-mannose (alpha-Man) specifically inhibits the adhesion of parasites to host cells suggest that the MBP plays a key role in the pathogenesis of Acanthamoeba infection by mediating host-parasite interactions. However, direct evidence showing that Acanthamoeba MBP is a virulence protein has been lacking. In this study, we demonstrate that the polyclonal immunoglobulin Y (IgY) antibodies prepared against affinity-purified Acanthamoeba MBP markedly inhibit the adhesion of parasites to host cells. The antibody also inhibited the Acanthamoeba-induced CPE on host cells. In contrast, preimmune IgY did not influence either the adhesion of the parasites to host cells or the amoeba-induced CPE. Using a variety of approaches, including affinity chromatography on an alpha-Man gel, electrophoresis under native and denaturing conditions, biotinylation of cell surface proteins, and immunostaining, it was conclusively established that Acanthamoeba MBP is located on the surface membranes of the parasites. Neutral-sugar analysis and lectin binding experiments using succinylated concanavalin A, a plant lectin with high affinity for mannose, revealed that Acanthamoeba MBP is itself a mannose-containing glycoprotein. N-Glycanase treatment to remove N-linked oligosaccharides shifted the subunit molecular mass of MBP from 130 kDa to 110 kDa. Hexosamine analysis revealed that Acanthamoeba MBP lacks detectable levels of GalNAc, suggesting the absence of O-linked oligosaccharides. In summary, we have characterized Acanthamoeba MBP and have shown that it is a major virulence protein responsible for host-parasite interactions and the parasite-induced target cell destruction.
Authors:
Marco Garate; Ibis Cubillos; Jeffrey Marchant; Noorjahan Panjwani
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Infection and immunity     Volume:  73     ISSN:  0019-9567     ISO Abbreviation:  Infect. Immun.     Publication Date:  2005 Sep 
Date Detail:
Created Date:  2005-08-22     Completed Date:  2005-10-12     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0246127     Medline TA:  Infect Immun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5775-81     Citation Subset:  IM    
Affiliation:
Department of Ophthalmology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA 02111, USA.
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MeSH Terms
Descriptor/Qualifier:
Acanthamoeba castellanii / immunology,  pathogenicity,  physiology*
Cell Adhesion / physiology
Cell Membrane / chemistry,  physiology
Cornea / parasitology,  physiology
Epithelial Cells / parasitology,  physiology
Humans
Mannose / chemistry
Mannose-Binding Lectin / chemistry*,  immunology,  physiology*
Protozoan Proteins / chemistry*,  physiology*
Virulence
Grant Support
ID/Acronym/Agency:
EY08706/EY/NEI NIH HHS; EY09349/EY/NEI NIH HHS; P30 EY13078/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/Mannose-Binding Lectin; 0/Protozoan Proteins; 31103-86-3/Mannose
Comments/Corrections

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