Trypanosoma brucei rhodesiense bloodstream forms: surface ricin-binding glycoproteins are localized exclusively in the flagellar pocket and the flagellar adhesion zone.
Specific binding of fluoresceinated succinyl-concanavalin A, wheat germ agglutinin, and ricin to untreated and trypsinized bloodstream forms of Trypanosoma brucei rhodesiense was quantitated by flow cytofluorimetry, and sites of lectin binding were identified by fluorescence microscopy. All three lectins only bound to the flagellar pocket of untreated parasites. When parasites were trypsinized to remove the variant surface glycoprotein coat, new lectin binding sites were exposed, and specific binding of all three lectins increased significantly. New specific binding sites for succinyl-concanavalin A and wheat germ agglutinin were present along both the free flagellum and flagellar adhesion zone and were uniformly distributed on the parasite surface. However, ricin did not bind uniformly on the surface and did not stain the free flagellum of trypsinized cells. Ricin only bound to the flagellar adhesion zone of trypsinized cells and of cells that had been treated with formaldehyde prior to staining. Electron microscopy of cells exposed to ricin-colloidal gold complexes revealed that that ricin binding was restricted to the anterior membrane of the flagellar pocket of untrypsinized cells and to this portion of the flagellar pocket and the cell body membrane in the flagellar adhesion zone of trypsinized cells. Evidence that these membranes constitute a functionally important membrane microdomain is reviewed.
Brickman
M J
MJ
Department of Microbiology and Immunology, Duke University School of Medicine, Durham, North Carolina 27710.
Balber
A E
AE
D000818:Animals; D001665:Binding Sites; D003208:Concanavalin A / Q000031:analogs & derivatives; D005434:Flow Cytometry; D037102:Lectins / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000378:metabolism; D008566:Membranes / Q000032:analysis / Q000378:metabolism / Q000648:ultrastructure; D009844:Oligosaccharides / Q000378:metabolism; D012276:Ricin / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000032:analysis* / Q000378:metabolism / Q000648:ultrastructure; D014357:Trypsin; D014909:Wheat Germ Agglutinins
D037102:Lectins; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A; C019558:succinylconcanavalin A; D012276:Ricin; D014357:Trypsin