Characterization of Lotus tetragonolobus fucolectin components for differences in hemagglutinating and macrophage activating activities.
The fucose binding proteins (FBP) extracted from Lotus tetragonolobus seeds were isolated by affinity chromatography and compared with affinity purified commercial preparations for physical, antigenic, and biological properties. All preparations contained three protein components as determined by polyacrylamide gel electrophoresis, each with a subunit molecular weight of approximately 27 Kd. FBP preparations were also found to be antigenically identical by immunodiffusion analysis and possessed similar biological activities for hemagglutination of group 0 erythrocytes and macrophage activation in the migration inhibition assay. A reversible temperature dependent hemagglutination characteristic was found; FBP agglutinated erythrocytes at 4 degrees and 22 degrees C but not at 37 degrees C, which was reversed by decreasing the incubation temperature from 37 degrees C to 22 degrees C. Temperature dependent binding of FBP for macrophages was also demonstrated. Adsorption of crude FBP by group 0 erythrocytes preferentially removed hemagglutinin without loss of macrophage activating properties. Similarly adsorption of FBP with macrophages preferentially removed macrophage activating component. Separation of the lectin components by DEAE cellulose chromatography yielded two major fractions: a potent hemagglutinin with weak macrophage activating properties and a potent macrophage activator with weak hemagglutinating activity. Separation of the crude lectin by ultrafiltration indicated that the macrophage activating component exists in a highly aggregated form which may determine its macrophage activating properties. Our results indicate that L. tetragonolobus consists of two distinct classes of components which correspond to tetrameric glycoproteins of 118-120 Kd with potent temperature dependent hemagglutinating activity and a highly aggregated dimeric component of 58 Kd with macrophage activating properties.
D000017:ABO Blood-Group System; D002848:Chromatography, DEAE-Cellulose; D004912:Erythrocytes / Q000276:immunology*; D005643:Fucose / Q000378:metabolism; D006388:Hemagglutinins / Q000302:isolation & purification*; D006801:Humans; D037102:Lectins / Q000276:immunology* / Q000302:isolation & purification; D008262:Macrophage Activation; D008264:Macrophages / Q000276:immunology*; D008970:Molecular Weight; D013696:Temperature; D014462:Ultrafiltration
D000017:ABO Blood-Group System; D006388:Hemagglutinins; D037102:Lectins; C027112:fucose-binding lectin; D005643:Fucose