Surface glycoproteins of differentiating neuroblastoma cells analyzed by lectin binding and flow cytometry.
Cell surface glycoproteins of mitotic neuroblastoma cells and cells differentiated by prostaglandin cyclic adenosine monophosphate treatment were quantified by flow cytometric analysis and specific fluorescent lectins. No differences in fluorescent lectin binding were seen between suspensions of mitotically active and differentiated N2AB-1 cells following exposure to either fluorescein (FL)-labeled soy bean agglutinin (FL-SBA) specific for N acetyl galactosamine or FL-concanavalin A (FL-CON A) which binds to mannose residues. These lectins, however, were shown to bind specifically to these cells as revealed by competitive blocking studies with hapten sugars. When FL Ulex europaeus (FL-UEA) specific for fucose was reacted with control or differentiated cells, no binding was seen even with an increased dose of lectin before or after enzyme treatment. However, differentiated N2AB-1 cells, reacted with FL-wheat germ agglutinin (FL-WGA) specific for N acetyl glucosamine, bound more FL-WGA than that seen for control cultures. Furthermore, specific sites for FL-WGA were shown to be saturable and were lost upon pretreatment of cells with neuraminidase. Neuraminidase pretreatment revealed masked sites for FL-CON A and FL-SBA since binding was increased at least twofold for these lectins on mitotic and differentiated cells. These data indicate that single cell measurements of surface glycoproteins can be made on living neural cells and that differentiation induces an increase in cell surface N-acetyl glucosamine residues.
Notter
M F
MF
Department of Neurobiology and Anatomy, University of Rochester, School of Medicine and Dentistry, New York 14642.
Leary
J F
JF
D000116:Acetylgalactosamine / Q000032:analysis / Q000378:metabolism; D000117:Acetylglucosamine / Q000032:analysis / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D003208:Concanavalin A / Q000378:metabolism; D005434:Flow Cytometry; D019793:Fluorescein; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006023:Glycoproteins / Q000032:analysis* / Q000378:metabolism; D037102:Lectins / Q000378:metabolism*; D008358:Mannose / Q000032:analysis / Q000378:metabolism; D051379:Mice; D009447:Neuroblastoma / Q000032:analysis*; D037121:Plant Lectins; D030262:Soybean Proteins; D014909:Wheat Germ Agglutinins / Q000378:metabolism; D010835:Phytohemagglutinins
D000116:Acetylgalactosamine; D000117:Acetylglucosamine; D003208:Concanavalin A; D019793:Fluorescein; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006023:Glycoproteins; D037102:Lectins; D008358:Mannose; D037121:Plant Lectins; D030262:Soybean Proteins; D014909:Wheat Germ Agglutinins; C028824:soybean lectin; D010835:Phytohemagglutinins